Sample records for laser scanning microscope

  1. ScanImage: flexible software for operating laser scanning microscopes.

    PubMed

    Pologruto, Thomas A; Sabatini, Bernardo L; Svoboda, Karel

    2003-05-17

    Laser scanning microscopy is a powerful tool for analyzing the structure and function of biological specimens. Although numerous commercial laser scanning microscopes exist, some of the more interesting and challenging applications demand custom design. A major impediment to custom design is the difficulty of building custom data acquisition hardware and writing the complex software required to run the laser scanning microscope. We describe a simple, software-based approach to operating a laser scanning microscope without the need for custom data acquisition hardware. Data acquisition and control of laser scanning are achieved through standard data acquisition boards. The entire burden of signal integration and image processing is placed on the CPU of the computer. We quantitate the effectiveness of our data acquisition and signal conditioning algorithm under a variety of conditions. We implement our approach in an open source software package (ScanImage) and describe its functionality. We present ScanImage, software to run a flexible laser scanning microscope that allows easy custom design.

  2. Purchase of a Laser Scanning Confocal Microscope at Xavier University of Louisiana

    DTIC Science & Technology

    2016-05-04

    SECURITY CLASSIFICATION OF: The purpose of this grant was to purchase a laser scanning confocal microscope to be used by multiple laboratories at...was being developed for undergraduate education. Over the course of the funding period, the microscope was purchased and installed, multiple training...Distribution Unlimited UU UU UU UU 04-05-2016 1-Feb-2015 31-Jan-2016 Final Report: Purchase of a Laser Scanning Confocal Microscope at Xavier

  3. Improved axial point spread function in a two-frequency laser scanning confocal fluorescence microscope

    NASA Astrophysics Data System (ADS)

    Wu, Jheng-Syong; Chung, Yung-Chin; Chien, Jun-Jei; Chou, Chien

    2018-01-01

    A two-frequency laser scanning confocal fluorescence microscope (TF-LSCFM) based on intensity modulated fluorescence signal detection was proposed. The specimen-induced spherical aberration and scattering effect were suppressed intrinsically, and high image contrast was presented due to heterodyne interference. An improved axial point spread function in a TF-LSCFM compared with a conventional laser scanning confocal fluorescence microscope was demonstrated and discussed.

  4. Scanning laser microscope for imaging nanostructured superconductors

    NASA Astrophysics Data System (ADS)

    Ishida, Takekazu; Arai, Kohei; Akita, Yukio; Miyanari, Mitsunori; Minami, Yusuke; Yotsuya, Tsutomu; Kato, Masaru; Satoh, Kazuo; Uno, Mayumi; Shimakage, Hisashi; Miki, Shigehito; Wang, Zhen

    2010-10-01

    The nanofabrication of superconductors yields various interesting features in superconducting properties. A variety of different imaging techniques have been developed for probing the local superconducting profiles. A scanning pulsed laser microscope has been developed by the combination of the XYZ piezo-driven stages and an optical fiber with an aspheric focusing lens. The scanning laser microscope is used to understand the position-dependent properties of a superconducting MgB 2 stripline of length 100 μm and width of 3 μm under constant bias current. Our results show that the superconducting stripline can clearly be seen in the contour image of the scanning laser microscope on the signal voltage. It is suggested from the observed image that the inhomogeneity is relevant in specifying the operating conditions such as detection efficiency of the sensor.

  5. The design and construction of a cost-efficient confocal laser scanning microscope

    NASA Astrophysics Data System (ADS)

    Xi, Peng; Rajwa, Bartlomiej; Jones, James T.; Robinson, J. Paul

    2007-03-01

    The optical dissection ability of confocal microscopy makes it a powerful tool for biological materials. However, the cost and complexity of confocal scanning laser microscopy hinders its wide application in education. We describe the construction of a simplified confocal scanning laser microscope and demonstrate three-dimensional projection based on cost-efficient commercial hardware, together with available open source software.

  6. Laser scanning confocal microscope with programmable amplitude, phase, and polarization of the illumination beam.

    PubMed

    Boruah, B R; Neil, M A A

    2009-01-01

    We describe the design and construction of a laser scanning confocal microscope with programmable beam forming optics. The amplitude, phase, and polarization of the laser beam used in the microscope can be controlled in real time with the help of a liquid crystal spatial light modulator, acting as a computer generated hologram, in conjunction with a polarizing beam splitter and two right angled prisms assembly. Two scan mirrors, comprising an on-axis fast moving scan mirror for line scanning and an off-axis slow moving scan mirror for frame scanning, configured in a way to minimize the movement of the scanned beam over the pupil plane of the microscope objective, form the XY scan unit. The confocal system, that incorporates the programmable beam forming unit and the scan unit, has been implemented to image in both reflected and fluorescence light from the specimen. Efficiency of the system to programmably generate custom defined vector beams has been demonstrated by generating a bottle structured focal volume, which in fact is the overlap of two cross polarized beams, that can simultaneously improve both the lateral and axial resolutions if used as the de-excitation beam in a stimulated emission depletion confocal microscope.

  7. A multiphoton laser scanning microscope setup for transcranial in vivo brain imaging on mice

    NASA Astrophysics Data System (ADS)

    Nase, Gabriele; Helm, P. Johannes; Reppen, Trond; Ottersen, Ole Petter

    2005-12-01

    We describe a multiphoton laser scanning microscope setup for transcranial in vivo brain imaging in mice. The modular system is based on a modified industrial standard Confocal Scanning Laser Microscope (CSLM) and is assembled mainly from commercially available components. A special multifunctional stage, which is optimized for both laser scanning microscopic observation and preparative animal surgery, has been developed and built. The detection unit includes a highly efficient photomultiplier tube installed in a Peltier-cooled thermal box shielding the detector from changes in room temperature and from distortions caused by external electromagnetic fields. The images are recorded using a 12-bit analog-to-digital converter. Depending on the characteristics of the staining, individual nerve cells can be imaged down to at least 100μm below the intact cranium and down to at least 200μm below the opened cranium.

  8. A study of cavity preparation by Er:YAG laser--observation of hard tooth structures by laser scanning microscope and examination of the time necessary to remove caries.

    PubMed

    Shigetani, Yoshimi; Okamoto, Akira; Abu-Bakr, Neamat; Iwaku, Masaaki

    2002-03-01

    The purpose of this study was to observe and measure the morphological changes that occur in the hard tissue after the application of Er:YAG laser. Another objective was to evaluate and compare the duration of application of both the laser apparatus and a conventional cutting device. In this study, sound and newly extracted carious tissues were used. The morphological changes in hard tooth structures produced by Er:YAG laser irradiation were examined by using a laser scanning microscope. Results showed that appropriate laser irradiation was 100 mJ/pulse for dentin, and 200 mJ/pulse for enamel. Also, the laser scanning microscope images were less damaged than the SEM images due to pretreatment of the specimens. The time taken to remove carious enamel by laser irradiation was slightly longer than the compared rotary cutting device; however, no differences between the two methods were observed in case of carious dentin removal.

  9. 75 FR 23272 - Government-Owned Inventions; Availability for Licensing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-03

    ...) Protection in Sunscreen Products Description of Invention: There are different types of ultraviolet (UV) rays..., PhD at 301-435-3131 or [email protected] for more information. Laser Scanning Microscopy for Three... data from a high-speed laser-scanning microscope and compute motion of the sample under the microscope...

  10. To boldly glow ... applications of laser scanning confocal microscopy in developmental biology.

    PubMed

    Paddock, S W

    1994-05-01

    The laser scanning confocal microscope (LSCM) is now established as an invaluable tool in developmental biology for improved light microscope imaging of fluorescently labelled eggs, embryos and developing tissues. The universal application of the LSCM in biomedical research has stimulated improvements to the microscopes themselves and the synthesis of novel probes for imaging biological structures and physiological processes. Moreover the ability of the LSCM to produce an optical series in perfect register has made computer 3-D reconstruction and analysis of light microscope images a practical option.

  11. A simple but precise method for quantitative measurement of the quality of the laser focus in a scanning optical microscope

    PubMed Central

    MACRAE, K.; TRAVIS, C.; AMOR, R.; NORRIS, G.; WILSON, S.H.; OPPO, G.‐L.; MCCONNELL, G.

    2015-01-01

    Summary We report a method for characterizing the focussing laser beam exiting the objective in a laser scanning microscope. This method provides the size of the optical focus, the divergence of the beam, the ellipticity and the astigmatism. We use a microscopic‐scale knife edge in the form of a simple transmission electron microscopy grid attached to a glass microscope slide, and a light‐collecting optical fibre and photodiode underneath the specimen. By scanning the laser spot from a reflective to a transmitting part of the grid, a beam profile in the form of an error function can be obtained and by repeating this with the knife edge at different axial positions relative to the beam waist, the divergence and astigmatism of the postobjective laser beam can be obtained. The measured divergence can be used to quantify how much of the full numerical aperture of the lens is used in practice. We present data of the beam radius, beam divergence, ellipticity and astigmatism obtained with low (0.15, 0.7) and high (1.3) numerical aperture lenses and lasers commonly used in confocal and multiphoton laser scanning microscopy. Our knife‐edge method has several advantages over alternative knife‐edge methods used in microscopy including that the knife edge is easy to prepare, that the beam can be characterized also directly under a cover slip, as necessary to reduce spherical aberrations for objectives designed to be used with a cover slip, and it is suitable for use with commercial laser scanning microscopes where access to the laser beam can be limited. PMID:25864964

  12. Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

    NASA Astrophysics Data System (ADS)

    Lorenzen, Manfred; Campbell, Duncan R.; Johnson, Craig W.

    1991-03-01

    Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner array for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.

  13. Optical fabrication and testing; Proceedings of the Meeting, Singapore, Oct. 22-27, 1990

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

    Lorenzen, M.; Campbell, D.R.; Johnson, C.W.

    1991-01-01

    Various papers on optical fabrication and testing are presented. Individual topics addressed include: interferometry with laser diodes, new methods for economic production of prisms and lenses, interferometer accuracy and precision, optical testing with wavelength scanning interferometer, digital Talbot interferometer, high-sensitivity interferometric technique for strain measurements, absolute interferometric testing of spherical surfaces, contouring using gratings created on an LCD panel, three-dimensional inspection using laser-based dynamic fringe projection, noncontact optical microtopography, laser scan microscope and infrared laser scan microscope, photon scanning tunneling microscopy. Also discussed are: combination-matching problems in the layout design of minilaser rangefinder, design and testing of a cube-corner arraymore » for laser ranging, mode and far-field pattern of diode laser-phased arrays, new glasses for optics and optoelectronics, optical properties of Li-doped ZnO films, application and machining of Zerodur for optical purposes, finish machining of optical components in mass production.« less

  14. Compensation of temporal and spatial dispersion for multiphoton acousto-optic laser-scanning microscopy

    NASA Astrophysics Data System (ADS)

    Iyer, Vijay; Saggau, Peter

    2003-10-01

    In laser-scanning microscopy, acousto-optic (AO) deflection provides a means to quickly position a laser beam to random locations throughout the field-of-view. Compared to conventional laser-scanning using galvanometer-driven mirrors, this approach increases the frame rate and signal-to-noise ratio, and reduces time spent illuminating sites of no interest. However, random-access AO scanning has not yet been combined with multi-photon microscopy, primarily because the femtosecond laser pulses employed are subject to significant amounts of both spatial and temporal dispersion upon propagation through common AO materials. Left uncompensated, spatial dispersion reduces the microscope"s spatial resolution while temporal dispersion reduces the multi-photon excitation efficacy. In previous work, we have demonstrated, 1) the efficacy of a single diffraction grating scheme which reduces the spatial dispersion at least 3-fold throughout the field-of-view, and 2) the use of a novel stacked-prism pre-chirper for compensating the temporal dispersion of a pair of AODs using a shorter mechanical path length (2-4X) than standard prism-pair arrangements. In this work, we demonstrate for the first time the use of these compensation approaches with a custom-made large-area slow-shear TeO2 AOD specifically suited for the development of a high-resolution 2-D random-access AO scanning multi-photon laser-scanning microscope (AO-MPLSM).

  15. Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope.

    PubMed

    Neděla, Vilém; Hřib, Jiří; Havel, Ladislav; Hudec, Jiří; Runštuk, Jiří

    2016-05-01

    This article describes the surface structure of Norway spruce early somatic embryos (ESEs) as a typical culture with asynchronous development. The microstructure of extracellular matrix covering ESEs were observed using the environmental scanning electron microscope as a primary tool and using the scanning electron microscope with cryo attachment and laser electron microscope as a complementary tool allowing our results to be proven independently. The fresh samples were observed in conditions of the air environment of the environmental scanning electron microscope (ESEM) with the pressure from 550Pa to 690Pa and the low temperature of the sample from -18°C to -22°C. The samples were studied using two different types of detector to allow studying either the thin surface structure or material composition. The scanning electron microscope with cryo attachment was used for imaging frozen extracellular matrix microstructure with higher resolution. The combination of both electron microscopy methods was suitable for observation of "native" plant samples, allowing correct evaluation of our results, free of error and artifacts. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. A setup for combined multiphoton laser scanning microscopic and multi-electrode patch clamp experiments on brain slices

    NASA Astrophysics Data System (ADS)

    Helm, P. Johannes; Reppen, Trond; Heggelund, Paul

    2009-02-01

    Multi Photon Laser Scanning Microscopy (MPLSM) appears today as one of the most powerful experimental tools in cellular neurophysiology, notably in studies of the functional dynamics of signal processing in single neurons. Simultaneous recording of fluorescence signals at high spatial and temporal resolution and electric signals by means of multi electrode patch clamp techniques have provided new paths for the systematic investigation of neuronal mechanisms. In particular, this approach has opened for direct studies of dendritic signal processing in neurons. We report about a setup optimized for simultaneous electrophysiological multi electrode patch clamp and multi photon laser scanning fluorescence microscopic experiments on brain slices. The microscopic system is based on a modified commercially available confocal scanning laser microscope (CLSM). From a technical and operational point of view, two developments are important: Firstly, in order to reduce the workload for the experimentalist, who in general is forced to concentrate on controlling the electrophysiological parameters during the recordings, a system of shutters has been installed together with dedicated electronic modules protecting the photo detectors against destructive light levels caused by erroneous opening or closing of microscopic light paths by the experimentalist. Secondly, the standard detection unit has been improved by installing the photomultiplier tubes (PMT) in a Peltier cooled thermal box shielding the detector from both room temperature and distortions caused by external electromagnetic fields. The electrophysiological system is based on an industrial standard multi patch clamp unit ergonomically arranged around the microscope stage. The electrophysiological and scanning processes can be time coordinated by standard trigger electronics.

  17. A high-resolution, confocal laser-scanning microscope and flash photolysis system for physiological studies.

    PubMed

    Parker, I; Callamaras, N; Wier, W G

    1997-06-01

    We describe the construction of a high-resolution confocal laser-scanning microscope, and illustrate its use for studying elementary Ca2+ signalling events in cells. An avalanche photodiode module and simple optical path provide a high efficiency system for detection of fluorescence signals, allowing use of a small confocal aperture giving near diffraction-limited spatial resolution (< 300 nm lateral and < 400 nm axial). When operated in line-scan mode, the maximum temporal resolution is 1 ms, and the associated computer software allows complete flexibility to record line-scans continuously for long (minutes) periods or to obtain any desired pixel resolution in x-y scans. An independent UV irradiation system permits simultaneous photolysis of caged compounds over either a uniform, wide field (arc lamp source) or at a tightly focussed spot (frequency-tripled Nd:YAG laser). The microscope thus provides a versatile tool for optical studies of dynamic cellular processes, as well as excellent resolution for morphological studies. The confocal scanner can be added to virtually any inverted microscope for a component cost that is only a small fraction of that of comparable commercial instruments, yet offers better performance and greater versatility.

  18. CONFOCAL LASER SCANNING MICROSCOPY OF RAT FOLLICLE DEVELOPMENT

    EPA Science Inventory

    This study used confocal laser scanning microscopy (CLSM) to study follicular development in millimeter pieces of rat ovary. To use this technology, it is essential to stain the tissue before laser excitation with the confocal microscope. Various fluorescent stains (Yo-Pro, Bo-Pr...

  19. Examination of silicon solar cells by means of the Scanning Laser Acoustic Microscope (SLAM)

    NASA Technical Reports Server (NTRS)

    Vorres, C.; Yuhas, D. E.

    1981-01-01

    The Scanning Laser Acoustic Microscope produces images of internal structure in materials. The acoustic microscope is an imaging system based upon acoustic rather than electromagnetic waves. Variations in the elastic propertis are primarily responsible for structure visualized in acoustic micrographs. The instrument used in these investigations is the SONOMICROSCOPE 100 which can be operated at ultrasonic frequencies of from 30 MHz to 500 MHz. The examination of the silicon solar cells was made at 100 MHz. Data are presented in the form of photomicrographs.

  20. Self-mixing laser diode included in scanning microwave microscope to the control of probe nanodisplacement

    NASA Astrophysics Data System (ADS)

    Usanov, D. A.; Skripal, A. V.; Astakhov, E. I.; Dobdin, S. Y.

    2018-04-01

    The possibilities of self-mixing interferometry for measuring nanodisplacement of a probe included in a near-field scanning microwave microscope have been considered. The features of the formation of a laser interference signal at current modulation of the wavelength of laser radiation have been investigated. Experimental responses of a semiconductor laser system included in scanning microwave microscope to control nanodisplacement of the probe have been demonstrated.To register the nanodisplacement of the probe, it is proposed to use the method of determining the stationary phase of a laser interference signal by low-frequency spectrum of a semiconductor laser. The change of the amplitudes of the spectral components in the spectrum of the interference signal due to creation of the standing wave in the external resonator of the laser self-mixing system has been shown. The form of the interference signal at current modulation of the radiation wavelength was experimentally obtained when the probe moves with a step of 80 nm. The results of measuring nanodisplacements of an electromagnetic translator STANDA 8MVT40-13 have been demonstrated. Deviation of the nanodisplacement of the proposed method does not exceed 15%.

  1. Parameters in selective laser melting for processing metallic powders

    NASA Astrophysics Data System (ADS)

    Kurzynowski, Tomasz; Chlebus, Edward; Kuźnicka, Bogumiła; Reiner, Jacek

    2012-03-01

    The paper presents results of studies on Selective Laser Melting. SLM is an additive manufacturing technology which may be used to process almost all metallic materials in the form of powder. Types of energy emission sources, mainly fiber lasers and/or Nd:YAG laser with similar characteristics and the wavelength of 1,06 - 1,08 microns, are provided primarily for processing metallic powder materials with high absorption of laser radiation. The paper presents results of selected variable parameters (laser power, scanning time, scanning strategy) and fixed parameters such as the protective atmosphere (argon, nitrogen, helium), temperature, type and shape of the powder material. The thematic scope is very broad, so the work was focused on optimizing the process of selective laser micrometallurgy for producing fully dense parts. The density is closely linked with other two conditions: discontinuity of the microstructure (microcracks) and stability (repeatability) of the process. Materials used for the research were stainless steel 316L (AISI), tool steel H13 (AISI), and titanium alloy Ti6Al7Nb (ISO 5832-11). Studies were performed with a scanning electron microscope, a light microscopes, a confocal microscope and a μCT scanner.

  2. Scanning thin-sheet laser imaging microscopy (sTSLIM) with structured illumination and HiLo background rejection.

    PubMed Central

    Schröter, Tobias J.; Johnson, Shane B.; John, Kerstin; Santi, Peter A.

    2011-01-01

    We report replacement of one side of a static illumination, dual sided, thin-sheet laser imaging microscope (TSLIM) with an intensity modulated laser scanner in order to implement structured illumination (SI) and HiLo image demodulation techniques for background rejection. The new system is equipped with one static and one scanned light-sheet and is called a scanning thin-sheet laser imaging microscope (sTSLIM). It is an optimized version of a light-sheet fluorescent microscope that is designed to image large specimens (<15 mm in diameter). In this paper we describe the hardware and software modifications to TSLIM that allow for static and uniform light-sheet illumination with SI and HiLo image demodulation. The static light-sheet has a thickness of 3.2 µm; whereas, the scanned side has a light-sheet thickness of 4.2 µm. The scanned side images specimens with subcellular resolution (<1 µm lateral and <4 µm axial resolution) with a size up to 15 mm. SI and HiLo produce superior contrast compared to both the uniform static and scanned light-sheets. HiLo contrast was greater than SI and is faster and more robust than SI because as it produces images in two-thirds of the time and exhibits fewer intensity streaking artifacts. PMID:22254177

  3. Effect of CO2 laser on Class V cavities of human molar teeth under a scanning electron microscope.

    PubMed

    Watanabe, I; Lopes, R A; Brugnera, A; Katayama, A Y; Gardini, A E

    1996-01-01

    The purpose of this study was to evaluate the effects of CO2 laser on dentin of class V cavities of extracted human molar teeth using a scanning electron microscope. SEM showed a smooth area with concentric lines formed by melting with subsequent recrystallization of dentin, areas of granulation, vitrified surface, numerous cracks, and irregular areas of descamative dentin. These data indicate that CO2 laser (4 and 6 watts) produces dentin alterations and limit its clinical applications.

  4. Recommendations for the design and the installation of large laser scanning microscopy systems

    NASA Astrophysics Data System (ADS)

    Helm, P. Johannes

    2012-03-01

    Laser Scanning Microscopy (LSM) has since the inventions of the Confocal Scanning Laser Microscope (CLSM) and the Multi Photon Laser Scanning Microscope (MPLSM) developed into an essential tool in contemporary life science and material science. The market provides an increasing number of turn-key and hands-off commercial LSM systems, un-problematic to purchase, set up and integrate even into minor research groups. However, the successful definition, financing, acquisition, installation and effective use of one or more large laser scanning microscopy systems, possibly of core facility character, often requires major efforts by senior staff members of large academic or industrial units. Here, a set of recommendations is presented, which are helpful during the process of establishing large systems for confocal or non-linear laser scanning microscopy as an effective operational resource in the scientific or industrial production process. Besides the description of technical difficulties and possible pitfalls, the article also illuminates some seemingly "less scientific" processes, i.e. the definition of specific laboratory demands, advertisement of the intention to purchase one or more large systems, evaluation of quotations, establishment of contracts and preparation of the local environment and laboratory infrastructure.

  5. Light and scanning electron microscope investigations comparing calculus removal using an Er:YAG laser and a frequency-doubled alexandrite laser

    NASA Astrophysics Data System (ADS)

    Rechmann, Peter; Hennig, Thomas; Sadegh, Hamid M. M.; Goldin, Dan S.

    1997-05-01

    With respect to lasers emitting within the mid-IR spectral domain fiber applicators are being developed. Intended is the use of these lasers in periodontal therapy and their application inside the gingival pocket. Aim of the study presented here is to compare the effect of an Er:YAG laser on dental calculus with the results following irradiation with a frequency doubled Alexandrite laser. The surface of freshly extracted wisdom teeth and of extracted teeth suffering from severe periodontitis were irradiated with both laser wavelengths using a standardized application protocol. Calculus on the enamel surface, at the enamel cementum junction and on the root surface was irradiated. For light microscope investigations undecalcified histological sections were prepared after treatment. For the scanning electron microscope teeth were dried in alcohol and sputtered with gold. Investigations revealed that with both laser systems calculus can be removed. Using the frequency doubled Alexandrite laser selective removal of calculus is possible while engaging the Er:YAG laser even at lowest energies necessary for calculus removal healthy cementum is ablated without control.

  6. Application of laser scanning speckle-microscopy for high-resolution express diagnostics of chlamydial infection

    NASA Astrophysics Data System (ADS)

    Ulyanov, Sergey; Larionova, Olga; Ulianova, Onega; Zaitsev, Sergey; Saltykov, Yury; Polyanina, Tatiana; Lyapina, Anna; Filonova, Nadezhda; Subbotina, Irina; Kalduzova, Irina; Utz, Sergey; Moiseeva, Yulia; Feodorova, Valentina

    2018-04-01

    Method of speckle-microscopy has been adapted to the problem of detection of Chlamydia trachomatis microbial cells in clinical samples. Prototype of laser scanning speckle-microscope has been designed. Spatial resolution and output characteristics of this microscope have been analyzed for the case of scanning of C. trachomatis bacteria inclusions - Elementary Bodies (EBs) inside the human cells, fixed on the glass. It has been demonstrated, that presence of C. trachomatis microbial cells in the sample can be easily detected using speckle microscopy.

  7. Solar-cell defect analyzer

    NASA Technical Reports Server (NTRS)

    Gauthier, M. K.; Miller, E. L.; Shumka, A.

    1980-01-01

    Laser-Scanning System pinpoints imperfections in solar cells. Entire solar panels containing large numbers of cells can be scanned. Although technique is similar to use of scanning electron microscope (SEM) to locate microscopic imperfections, it differs in that large areas may be examined, including entire solar panels, and it is not necessary to remove cover glass or encapsulants.

  8. Characterization of a subwavelength-scale 3D void structure using the FDTD-based confocal laser scanning microscopic image mapping technique.

    PubMed

    Choi, Kyongsik; Chon, James W; Gu, Min; Lee, Byoungho

    2007-08-20

    In this paper, a simple confocal laser scanning microscopic (CLSM) image mapping technique based on the finite-difference time domain (FDTD) calculation has been proposed and evaluated for characterization of a subwavelength-scale three-dimensional (3D) void structure fabricated inside polymer matrix. The FDTD simulation method adopts a focused Gaussian beam incident wave, Berenger's perfectly matched layer absorbing boundary condition, and the angular spectrum analysis method. Through the well matched simulation and experimental results of the xz-scanned 3D void structure, we first characterize the exact position and the topological shape factor of the subwavelength-scale void structure, which was fabricated by a tightly focused ultrashort pulse laser. The proposed CLSM image mapping technique based on the FDTD can be widely applied from the 3D near-field microscopic imaging, optical trapping, and evanescent wave phenomenon to the state-of-the-art bio- and nanophotonics.

  9. Two-photon microscopy and spectroscopy based on a compact confocal scanning head

    NASA Astrophysics Data System (ADS)

    Diaspro, Alberto; Chirico, Giberto; Federici, Federico; Cannone, Fabio; Beretta, Sabrina; Robello, Mauro; Olivini, Francesca; Ramoino, Paola

    2001-07-01

    We have combined a confocal laser scanning head modified for TPE (two-photon excitation) microscopy with some spectroscopic modules to study single molecules and molecular aggregates. The behavior of the TPE microscope unit has been characterized by means of point spread function measurements and of the demonstration of its micropatterning abilities. One-photon and two-photon mode can be simply accomplished by switching from a mono-mode optical fiber (one-photon) coupled to conventional laser sources to an optical module that allows IR laser beam (two- photon/TPE) delivery to the confocal laser scanning head. We have then described the characterization of the two-photon microscope for spectroscopic applications: fluorescence correlation, lifetime and fluorescence polarization anisotropy measurements. We describe the measurement of the response of the two-photon microscope to the light polarization and discuss fluorescence polarization anisotropy measurements on Rhodamine 6G as a function of the viscosity and on a globular protein, the Beta-lactoglobulin B labeled with Alexa 532 at very high dilutions. The average rotational and translational diffusion coefficients measured with fluorescence polarization anisotropy and fluorescence correlation methods are in good agreement with the protein size, therefore validating the use of the microscope for two-photon spectroscopy on biomolecules.

  10. Scanning thin-sheet laser imaging microscopy (sTSLIM) with structured illumination and HiLo background rejection.

    PubMed

    Schröter, Tobias J; Johnson, Shane B; John, Kerstin; Santi, Peter A

    2012-01-01

    We report replacement of one side of a static illumination, dual sided, thin-sheet laser imaging microscope (TSLIM) with an intensity modulated laser scanner in order to implement structured illumination (SI) and HiLo image demodulation techniques for background rejection. The new system is equipped with one static and one scanned light-sheet and is called a scanning thin-sheet laser imaging microscope (sTSLIM). It is an optimized version of a light-sheet fluorescent microscope that is designed to image large specimens (<15 mm in diameter). In this paper we describe the hardware and software modifications to TSLIM that allow for static and uniform light-sheet illumination with SI and HiLo image demodulation. The static light-sheet has a thickness of 3.2 µm; whereas, the scanned side has a light-sheet thickness of 4.2 µm. The scanned side images specimens with subcellular resolution (<1 µm lateral and <4 µm axial resolution) with a size up to 15 mm. SI and HiLo produce superior contrast compared to both the uniform static and scanned light-sheets. HiLo contrast was greater than SI and is faster and more robust than SI because as it produces images in two-thirds of the time and exhibits fewer intensity streaking artifacts. 2011 Optical Society of America

  11. Handheld nonlinear microscope system comprising a 2 MHz repetition rate, mode-locked Yb-fiber laser for in vivo biomedical imaging

    PubMed Central

    Krolopp, Ádám; Csákányi, Attila; Haluszka, Dóra; Csáti, Dániel; Vass, Lajos; Kolonics, Attila; Wikonkál, Norbert; Szipőcs, Róbert

    2016-01-01

    A novel, Yb-fiber laser based, handheld 2PEF/SHG microscope imaging system is introduced. It is suitable for in vivo imaging of murine skin at an average power level as low as 5 mW at 200 kHz sampling rate. Amplified and compressed laser pulses having a spectral bandwidth of 8 to 12 nm at around 1030 nm excite the biological samples at a ~1.89 MHz repetition rate, which explains how the high quality two-photon excitation fluorescence (2PEF) and second harmonic generation (SHG) images are obtained at the average power level of a laser pointer. The scanning, imaging and detection head, which comprises a conventional microscope objective for beam focusing, has a physical length of ~180 mm owing to the custom designed imaging telescope system between the laser scanner mirrors and the entrance aperture of the microscope objective. Operation of the all-fiber, all-normal dispersion Yb-fiber ring laser oscillator is electronically controlled by a two-channel polarization controller for Q-switching free mode-locked operation. The whole nonlinear microscope imaging system has the main advantages of the low price of the fs laser applied, fiber optics flexibility, a relatively small, light-weight scanning and detection head, and a very low risk of thermal or photochemical damage of the skin samples. PMID:27699118

  12. Handheld nonlinear microscope system comprising a 2 MHz repetition rate, mode-locked Yb-fiber laser for in vivo biomedical imaging.

    PubMed

    Krolopp, Ádám; Csákányi, Attila; Haluszka, Dóra; Csáti, Dániel; Vass, Lajos; Kolonics, Attila; Wikonkál, Norbert; Szipőcs, Róbert

    2016-09-01

    A novel, Yb-fiber laser based, handheld 2PEF/SHG microscope imaging system is introduced. It is suitable for in vivo imaging of murine skin at an average power level as low as 5 mW at 200 kHz sampling rate. Amplified and compressed laser pulses having a spectral bandwidth of 8 to 12 nm at around 1030 nm excite the biological samples at a ~1.89 MHz repetition rate, which explains how the high quality two-photon excitation fluorescence (2PEF) and second harmonic generation (SHG) images are obtained at the average power level of a laser pointer. The scanning, imaging and detection head, which comprises a conventional microscope objective for beam focusing, has a physical length of ~180 mm owing to the custom designed imaging telescope system between the laser scanner mirrors and the entrance aperture of the microscope objective. Operation of the all-fiber, all-normal dispersion Yb-fiber ring laser oscillator is electronically controlled by a two-channel polarization controller for Q-switching free mode-locked operation. The whole nonlinear microscope imaging system has the main advantages of the low price of the fs laser applied, fiber optics flexibility, a relatively small, light-weight scanning and detection head, and a very low risk of thermal or photochemical damage of the skin samples.

  13. Intensity calibration of a laser scanning confocal microscope based on concentrated dyes.

    PubMed

    Model, Michael A; Blank, James L

    2006-10-01

    To find water-soluble fluorescent dyes with absorption in various regions of the spectrum and investigate their utility as standards for laser scanning confocal microscopy. Several dyes were found to have characteristics required for fluorescence microscopy standards. The intensity of biological fluorescent specimens was measured against the emission of concentrated dyes. Results using different optics and different microscopes were compared. Slides based on concentrated dyes can be prepared in a highly reproducible manner and are stable under laser scanning. Normalized fluorescence of biological specimens remains consistent with different objective lenses and is tolerant to some mismatch in optical filters or imperfect pinhole alignment. Careful choice of scanning parameters is necessary to ensure linearity of intensity measurements. Concentrated dyes provide a robust and inexpensive intensity standard that can be used in basic research or clinical studies.

  14. Two-photon laser scanning microscopy with electrowetting-based prism scanning

    PubMed Central

    Supekar, Omkar D.; Ozbay, Baris N.; Zohrabi, Mo; Nystrom, Philip D.; Futia, Gregory L.; Restrepo, Diego; Gibson, Emily A.; Gopinath, Juliet T.; Bright, Victor M.

    2017-01-01

    Laser scanners are an integral part of high resolution biomedical imaging systems such as confocal or 2-photon excitation (2PE) microscopes. In this work, we demonstrate the utility of electrowetting on dielectric (EWOD) prisms as a lateral laser-scanning element integrated in a conventional 2PE microscope. To the best of our knowledge, this is the first such demonstration for EWOD prisms. EWOD devices provide a transmissive, low power consuming, and compact alternative to conventional adaptive optics, and hence this technology has tremendous potential. We demonstrate 2PE microscope imaging of cultured mouse hippocampal neurons with a FOV of 130 × 130 μm2 using EWOD prism scanning. In addition, we show simulations of the optical system with the EWOD prism, to evaluate the effect of propagating a Gaussian beam through the EWOD prism on the imaging quality. Based on the simulation results a beam size of 0.91 mm full width half max was chosen to conduct the imaging experiments, resulting in a numerical aperture of 0.17 of the imaging system. PMID:29296477

  15. Frequency-doubled Alexandrite laser for use in periodontology: a scanning electron microscopic investigation

    NASA Astrophysics Data System (ADS)

    Rechmann, Peter; Hennig, Thomas

    1996-12-01

    During prior studies it could be demonstrated that engaging a frequency double Alexandrite-laser allows a fast and strictly selective ablation of supra- and subgingival calculus. Furthermore, the removal of unstained microbial plaque was observed. First conclusions were drawn following light microscopic investigations on undecalcified sections of irradiated teeth. In the present study the cementum surface after irradiation with a frequency doubled Alexandrite-laser was observed by means of a scanning electron microscope. After irradiation sections of teeth were dried in alcohol and sputtered with gold. In comparison irradiated cementum surfaces of unerupted operatively removed wisdom teeth and tooth surfaces after the selective removal of calculus were investigated. A complete removal of calculus was observed as well as a remaining smooth surface of irradiated cementum.

  16. Spectral ophthalmoscopy based on supercontinuum

    NASA Astrophysics Data System (ADS)

    Cheng, Yueh-Hung; Yu, Jiun-Yann; Wu, Han-Hsuan; Huang, Bo-Jyun; Chu, Shi-Wei

    2010-02-01

    Confocal scanning laser ophthalmoscope (CSLO) has been established to be an important diagnostic tool for retinopathies like age-related macular degeneration, glaucoma and diabetes. Compared to a confocal laser scanning microscope, CSLO is also capable of providing optical sectioning on retina with the aid of a pinhole, but the microscope objective is replaced by the optics of eye. Since optical spectrum is the fingerprint of local chemical composition, it is attractive to incorporate spectral acquisition into CSLO. However, due to the limitation of laser bandwidth and chromatic/geometric aberration, the scanning systems in current CSLO are not compatible with spectral imaging. Here we demonstrate a spectral CSLO by combining a diffraction-limited broadband scanning system and a supercontinuum laser source. Both optical sectioning capability and sub-cellular resolution are demonstrated on zebrafish's retina. To our knowledge, it is also the first time that CSLO is applied onto the study of fish vision. The versatile spectral CSLO system will be useful to retinopathy diagnosis and neuroscience research.

  17. Hyperspectral imaging with laser-scanning sum-frequency generation microscopy

    PubMed Central

    Hanninen, Adam; Shu, Ming Wai; Potma, Eric O.

    2017-01-01

    Vibrationally sensitive sum-frequency generation (SFG) microscopy is a chemically selective imaging technique sensitive to non-centrosymmetric molecular arrangements in biological samples. The routine use of SFG microscopy has been hampered by the difficulty of integrating the required mid-infrared excitation light into a conventional, laser-scanning nonlinear optical (NLO) microscope. In this work, we describe minor modifications to a regular laser-scanning microscope to accommodate SFG microscopy as an imaging modality. We achieve vibrationally sensitive SFG imaging of biological samples with sub-μm resolution at image acquisition rates of 1 frame/s, almost two orders of magnitude faster than attained with previous point-scanning SFG microscopes. Using the fast scanning capability, we demonstrate hyperspectral SFG imaging in the CH-stretching vibrational range and point out its use in the study of molecular orientation and arrangement in biologically relevant samples. We also show multimodal imaging by combining SFG microscopy with second-harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) on the same imaging platfrom. This development underlines that SFG microscopy is a unique modality with a spatial resolution and image acquisition time comparable to that of other NLO imaging techniques, making point-scanning SFG microscopy a valuable member of the NLO imaging family. PMID:28966861

  18. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: LASER POWER MEASUREMENTS

    EPA Science Inventory

    Laser power abstract
    The reliability of the confocal laser-scanning microscope (CLSM) to obtain intensity measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. The laser power test appears to be one ...

  19. Apertureless scanning microscope probe as a detector of semiconductor laser emission

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

    Dunaevskiy, Mikhail, E-mail: Mike.Dunaeffsky@mail.ioffe.ru; National Research University of Information Technologies, Mechanics and Optics; Dontsov, Anton

    2015-04-27

    An operating semiconductor laser has been studied using a scanning probe microscope. A shift of the resonance frequency of probe that is due to its heating by laser radiation has been analyzed. The observed shift is proportional to the absorbed radiation and can be used to measure the laser near field or its output power. A periodical dependence of the measured signal has been observed as a function of distance between the probe and the surface of the laser due to the interference of the outgoing and cantilever-reflected waves. Due to the multiple reflections resulting in the interference, the lightmore » absorption by the probe cantilever is greatly enhanced compared with a single pass case. Interaction of infrared emission of a diode laser with different probes has been studied.« less

  20. Easy performance of 6-color confocal immunofluorescence with 4-laser line microscopes.

    PubMed

    Eissing, Nathalie; Heger, Lukas; Baranska, Anna; Cesnjevar, Robert; Büttner-Herold, Maike; Söder, Stephan; Hartmann, Arndt; Heidkamp, Gordon F; Dudziak, Diana

    2014-09-01

    Confocal laser scanning microscopy is an advanced technique for imaging tissue samples in vitro and in vivo at high optical resolution. The development of new fluorochrome variants do not only make it possible to perform multicolor flow cytometry of single cells, but in combination with high resolution laser scanning systems also to investigate the distribution of cells in lymphoid tissues by confocal immunofluorescence analyses, thus allowing the distinction of various cell populations directly in the tissue. Here, we provide a protocol for the visualization of at least six differently fluorochrome-labeled antibodies at the same time using a conventional confocal laser scanning microscope with four laser lines (405 nm, 488 nm, 555 nm, and 639 nm laser wavelength) in both murine and human tissue samples. We further demonstrate that compensation correction algorithms are not necessary to reduce spillover of fluorochromes into other channels when the used fluorochromes are combined according to their specific emission bands and the varying Stokes shift for co-excited fluorochromes with the same laser line. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Use of digital micromirror devices as dynamic pinhole arrays for adaptive confocal fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Pozzi, Paolo; Wilding, Dean; Soloviev, Oleg; Vdovin, Gleb; Verhaegen, Michel

    2018-02-01

    In this work, we present a new confocal laser scanning microscope capable to perform sensorless wavefront optimization in real time. The device is a parallelized laser scanning microscope in which the excitation light is structured in a lattice of spots by a spatial light modulator, while a deformable mirror provides aberration correction and scanning. A binary DMD is positioned in an image plane of the detection optical path, acting as a dynamic array of reflective confocal pinholes, images by a high performance cmos camera. A second camera detects images of the light rejected by the pinholes for sensorless aberration correction.

  2. Nano-material processing with laser radiation in the near field of a scanning probe tip

    NASA Astrophysics Data System (ADS)

    Jersch, J.; Demming, F.; Hildenhagen, J.; Dickmann, K.

    1998-04-01

    We report preliminary results of using a scanning probe microscope/laser combination to perform nanostructuring on insulator and metal surfaces in air. This technique enables processing of structures with a lateral resolution of approximately 10 nm. In this paper we present our last structuring results with both scanning tunnelling and scanning force microscopy. Some possible interaction mechanisms responsible for the structuring will be discussed.

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

    Tsai, Ming-Hung; School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei 110, Taiwan; Haung, Chiung-Fang

    In this study, neodymium-doped yttrium orthovanadate (Nd:YVO{sub 4}) as a laser source with different scanning speeds was used on biomedical Ti surface. The microstructural and biological properties of laser-modified samples were investigated by means of optical microscope, electron microscope, X-ray diffraction, surface roughness instrument, contact angle and cell cytotoxicity assay. After laser modification, the rough volcano-like recast layer with micro-/nanoporous structure and wave-like recast layer with nanoporous structure were generated on the surfaces of laser-modified samples, respectively. It was also found out that, an α → (α + rutile-TiO{sub 2}) phase transition occurred on the recast layers of laser-modified samples.more » The Ti surface becomes hydrophilic at a high speed laser scanning. Moreover, the cell cytotoxicity assay demonstrated that laser-modified samples did not influence the cell adhesion and proliferation behaviors of osteoblast (MG-63) cell. The laser with 50 mm/s scanning speed induced formation of rough volcano-like recast layer accompanied with micro-/nanoporous structure, which can promote cell adhesion and proliferation of MG-63 cell on Ti surface. The results indicated that the laser treatment was a potential technology to enhance the biocompatibility for titanium. - Highlights: • Laser induced the formation of recast layer with micro-/nanoporous structure on Ti. • An α → (α + rutile-TiO{sub 2}) phase transition was observed within the recast layer. • The Ti surface becomes hydrophilic at a high speed laser scanning. • Laser-modified samples exhibit good biocompatibility to osteoblast (MG-63) cell.« less

  4. Confocal laser scanning microscopic photoconversion: a new method to stabilize fluorescently labeled cellular elements for electron microscopic analysis.

    PubMed

    Colello, Raymond J; Tozer, Jordan; Henderson, Scott C

    2012-01-01

    Photoconversion, the method by which a fluorescent dye is transformed into a stable, osmiophilic product that can be visualized by electron microscopy, is the most widely used method to enable the ultrastructural analysis of fluorescently labeled cellular structures. Nevertheless, the conventional method of photoconversion using widefield fluorescence microscopy requires long reaction times and results in low-resolution cell targeting. Accordingly, we have developed a photoconversion method that ameliorates these limitations by adapting confocal laser scanning microscopy to the procedure. We have found that this method greatly reduces photoconversion times, as compared to conventional wide field microscopy. Moreover, region-of-interest scanning capabilities of a confocal microscope facilitate the targeting of the photoconversion process to individual cellular or subcellular elements within a fluorescent field. This reduces the area of the cell exposed to light energy, thereby reducing the ultrastructural damage common to this process when widefield microscopes are employed. © 2012 by John Wiley & Sons, Inc.

  5. Laser Scanning In Inspection

    NASA Astrophysics Data System (ADS)

    West, Patricia; Baker, Lionel R.

    1989-03-01

    This paper is a review of the applications of laser scanning in inspection. The reasons for the choice of a laser in flying spot scanning and the optical properties of a laser beam which are of value in a scanning instrument will be given. The many methods of scanning laser beams in both one and two dimensions will be described. The use of one dimensional laser scanners for automatic surface inspection for transmitting and reflective products will be covered in detail, with particular emphasis on light collection techniques. On-line inspection applications which will be mentioned include: photographic film web, metal strip products, paper web, glass sheet, car body paint surfaces and internal cylinder bores. Two dimensional laser scanning is employed in applications where increased resolution, increased depth of focus, and better contrast are required compared with conventional vidicon TV or solid state array cameras. Such examples as special microscope laser scanning systems and a TV compatible system for use in restricted areas of a nuclear reactor will be described. The technical and economic benefits and limitations of laser scanning video systems will be compared with conventional TV and CCD array devices.

  6. Femtosecond pulse laser-oriented recording on dental prostheses: a trial introduction.

    PubMed

    Ichikawa, Tetsuo; Hayasaki, Yoshio; Fujita, Keiji; Nagao, Kan; Murata, Masayo; Kawano, Takanori; Chen, JianRong

    2006-12-01

    The purpose of this study was to evaluate the feasibility of using a femtosecond pulse laser processing technique to store information on a dental prosthesis. Commercially pure titanium plates were processed by a femtosecond pulse laser system. The processed surface structure was observed with a reflective illumination microscope, scanning electron microscope, and atomic force microscope. Processed area was an almost conical pit with a clear boundary. When laser pulse energy was 2 microJ, the diameter and depth were approximately 10microm and 0.2 microm respectively--whereby both increased with laser pulse energy. Further, depth of pit increased with laser pulse number without any thermal effect. This study showed that the femtosecond pulse processing system was capable of recording personal identification and optional additional information on a dental prosthesis.

  7. Lateral resolution testing of a novel developed confocal microscopic imaging system

    NASA Astrophysics Data System (ADS)

    Zhang, Xin; Zhang, Yunhai; Chang, Jian; Huang, Wei; Xue, Xiaojun; Xiao, Yun

    2015-10-01

    Laser scanning confocal microscope has been widely used in biology, medicine and material science owing to its advantages of high resolution and tomographic imaging. Based on a set of confirmatory experiments and system design, a novel confocal microscopic imaging system is developed. The system is composed of a conventional fluorescence microscope and a confocal scanning unit. In the scanning unit a laser beam coupling module provides four different wavelengths 405nm 488nm 561nm and 638nm which can excite a variety of dyes. The system works in spot-to-spot scanning mode with a two-dimensional galvanometer. A 50 microns pinhole is used to guarantee that stray light is blocked and only the fluorescence signal from the focal point can be received . The three-channel spectral splitter is used to perform fluorescence imaging at three different working wavelengths simultaneously. The rat kidney tissue slice is imaged using the developed confocal microscopic imaging system. Nucleues labeled by DAPI and kidney spherule curved pipe labeled by Alexa Fluor 488 can be imaged clearly and respectively, realizing the distinction between the different components of mouse kidney tissue. The three-dimensional tomographic imaging of mouse kidney tissue is reconstructed by several two-dimensional images obtained in different depths. At last the resolution of the confocal microscopic imaging system is tested quantitatively. The experimental result shows that the system can achieve lateral resolution priority to 230nm.

  8. Laser-assisted electron tunneling in a STM junction

    NASA Astrophysics Data System (ADS)

    Chang, Shunhua Thomas

    2000-10-01

    Since its introduction in 1981, the Nobel prize-winning scanning tunneling microscope (STM) has been developed into a powerful yet conceptually simple instrument, replacing traditional scanning and transmission electron microscopes (SEM/TEM) in many of the microscopic surface phenomenon studies. The strength of the STM stems from the sensitive tunneling current-potential barrier width relationship of the electron tunneling process, and has been used to re-examine the frequency-mixing and harmonic generation properties of an non-linear metal- oxide-metal (MOM) tunneling junction. In this research, electron-tunneling events under polarized laser radiation at 514.5-nm argon and 10.6-μm carbon dioxide laser wavelengths were investigated. The objective is to understand the underlying interactive mechanisms between the tunneling junction and the external laser excitation. A commercial scanning tunneling microscope head and controller were incorporated into the experimental setup. Operation characteristics and the electrical properties of the STM junction were determined. Tunneling current and distance responses with respect to different laser polarization, modulation frequency, incident power, and tunneling distance were also conducted. From the experimental results it is shown that thermal expansion effect was the dominant source of response for laser modulation frequency up to about 100 kHz, in quantitative agreement with theoretical calculations. Different laser polarizations as the experiments demonstrated did not contribute significantly to the STM response in the investigated frequency range. The electric field induced by the laser beam was calculated to be one to two order of magnitudes lower than the field required to initiate field emission where the tunneling junction I- V curve is most non-linear. Also, the electrical coupling of the incident laser at the STM junction was determined to be non-critical at visible laser wavelength, and the reflected laser energy from the sample re-entering the junction was shown to be weak and did not influence the ongoing electron tunneling process. In conclusion, the thermal expansion of the physical tunneling junction was found to be responsible to the tunneling current modulation in a laser - STM setup for laser modulation frequencies in the lower frequency range.

  9. Note: Laser beam scanning using a ferroelectric liquid crystal spatial light modulator

    NASA Astrophysics Data System (ADS)

    Das, Abhijit; Boruah, Bosanta R.

    2014-04-01

    In this work we describe laser beam scanning using a ferroelectric liquid crystal spatial light modulator. Commercially available ferroelectric liquid crystal spatial light modulators are capable of displaying 85 colored images in 1 s using a time dithering technique. Each colored image, in fact, comprises 24 single bit (black and white) images displayed sequentially. We have used each single bit image to write a binary phase hologram. For a collimated laser beam incident on the hologram, one of the diffracted beams can be made to travel along a user defined direction. We have constructed a beam scanner employing the above arrangement and demonstrated its use to scan a single laser beam in a laser scanning optical sectioning microscope setup.

  10. Two-Photon Imaging with Diffractive Optical Elements

    PubMed Central

    Watson, Brendon O.; Nikolenko, Volodymyr; Yuste, Rafael

    2009-01-01

    Two-photon imaging has become a useful tool for optical monitoring of neural circuits, but it requires high laser power and serial scanning of each pixel in a sample. This results in slow imaging rates, limiting the measurements of fast signals such as neuronal activity. To improve the speed and signal-to-noise ratio of two-photon imaging, we introduce a simple modification of a two-photon microscope, using a diffractive optical element (DOE) which splits the laser beam into several beamlets that can simultaneously scan the sample. We demonstrate the advantages of DOE scanning by enhancing the speed and sensitivity of two-photon calcium imaging of action potentials in neurons from neocortical brain slices. DOE scanning can easily improve the detection of time-varying signals in two-photon and other non-linear microscopic techniques. PMID:19636390

  11. Effects of conventional welding and laser welding on the tensile strength, ultimate tensile strength and surface characteristics of two cobalt-chromium alloys: a comparative study.

    PubMed

    Madhan Kumar, Seenivasan; Sethumadhava, Jayesh Raghavendra; Anand Kumar, Vaidyanathan; Manita, Grover

    2012-06-01

    The purpose of this study was to evaluate the efficacy of laser welding and conventional welding on the tensile strength and ultimate tensile strength of the cobalt-chromium alloy. Samples were prepared with two commercially available cobalt-chromium alloys (Wironium plus and Diadur alloy). The samples were sectioned and the broken fragments were joined using Conventional and Laser welding techniques. The welded joints were subjected to tensile and ultimate tensile strength testing; and scanning electron microscope to evaluate the surface characteristics at the welded site. Both on laser welding as well as on conventional welding technique, Diadur alloy samples showed lesser values when tested for tensile and ultimate tensile strength when compared to Wironium alloy samples. Under the scanning electron microscope, the laser welded joints show uniform welding and continuous molt pool all over the surface with less porosity than the conventionally welded joints. Laser welding is an advantageous method of connecting or repairing cast metal prosthetic frameworks.

  12. A high-resolution combined scanning laser and widefield polarizing microscope for imaging at temperatures from 4 K to 300 K.

    PubMed

    Lange, M; Guénon, S; Lever, F; Kleiner, R; Koelle, D

    2017-12-01

    Polarized light microscopy, as a contrast-enhancing technique for optically anisotropic materials, is a method well suited for the investigation of a wide variety of effects in solid-state physics, as, for example, birefringence in crystals or the magneto-optical Kerr effect (MOKE). We present a microscopy setup that combines a widefield microscope and a confocal scanning laser microscope with polarization-sensitive detectors. By using a high numerical aperture objective, a spatial resolution of about 240 nm at a wavelength of 405 nm is achieved. The sample is mounted on a 4 He continuous flow cryostat providing a temperature range between 4 K and 300 K, and electromagnets are used to apply magnetic fields of up to 800 mT with variable in-plane orientation and 20 mT with out-of-plane orientation. Typical applications of the polarizing microscope are the imaging of the in-plane and out-of-plane magnetization via the longitudinal and polar MOKE, imaging of magnetic flux structures in superconductors covered with a magneto-optical indicator film via the Faraday effect, or imaging of structural features, such as twin-walls in tetragonal SrTiO 3 . The scanning laser microscope furthermore offers the possibility to gain local information on electric transport properties of a sample by detecting the beam-induced voltage change across a current-biased sample. This combination of magnetic, structural, and electric imaging capabilities makes the microscope a viable tool for research in the fields of oxide electronics, spintronics, magnetism, and superconductivity.

  13. A high-resolution combined scanning laser and widefield polarizing microscope for imaging at temperatures from 4 K to 300 K

    NASA Astrophysics Data System (ADS)

    Lange, M.; Guénon, S.; Lever, F.; Kleiner, R.; Koelle, D.

    2017-12-01

    Polarized light microscopy, as a contrast-enhancing technique for optically anisotropic materials, is a method well suited for the investigation of a wide variety of effects in solid-state physics, as, for example, birefringence in crystals or the magneto-optical Kerr effect (MOKE). We present a microscopy setup that combines a widefield microscope and a confocal scanning laser microscope with polarization-sensitive detectors. By using a high numerical aperture objective, a spatial resolution of about 240 nm at a wavelength of 405 nm is achieved. The sample is mounted on a 4He continuous flow cryostat providing a temperature range between 4 K and 300 K, and electromagnets are used to apply magnetic fields of up to 800 mT with variable in-plane orientation and 20 mT with out-of-plane orientation. Typical applications of the polarizing microscope are the imaging of the in-plane and out-of-plane magnetization via the longitudinal and polar MOKE, imaging of magnetic flux structures in superconductors covered with a magneto-optical indicator film via the Faraday effect, or imaging of structural features, such as twin-walls in tetragonal SrTiO3. The scanning laser microscope furthermore offers the possibility to gain local information on electric transport properties of a sample by detecting the beam-induced voltage change across a current-biased sample. This combination of magnetic, structural, and electric imaging capabilities makes the microscope a viable tool for research in the fields of oxide electronics, spintronics, magnetism, and superconductivity.

  14. In-situ investigation of laser surface modifications of WC-Co hard metals inside a scanning electron microscope

    NASA Astrophysics Data System (ADS)

    Mueller, H.; Wetzig, K.; Schultrich, B.; Pompe, Wolfgang; Chapliev, N. I.; Konov, Vitaly I.; Pimenov, S. M.; Prokhorov, Alexander M.

    1989-05-01

    The investigation of laser interaction with solid surfaces and of the resulting mechanism of surface modification are of technical interest to optimize technological processes, and they are also of fundamental scientific importance. Most instructive indormation is available with the ail of the in-situ techniques. For instance, measuring of the photon emission of the irradiated surface ane the plasma torch (if it is produced) simultaneously to laser action, makes it possible to gain a global characterization of the laser-solid interaction. In order to obtain additional information about surface and structure modifications in microscopic detail , a laser and scanning electron microscope were combined in to a tandem equipment (LASEM). Inside this eqiipment the microscopic observation is carried out directly at the laser irradiated area without any displacement of the sample. In this way, the stepwise development of surface modification during multipulse irradiation is visible in microscopic details and much more reliable information about the surface modification process is obtainable in comparison to an external laser irradiation. Such kind of equipments were realized simultaneously and independently in the Institut of General Physics (Moscow) and the Central Institute of Solid State Physics and Material Research (Dresden) using a CO2 and a LTd-glass-laser, respectively. In the following the advantages and possibilities of a LASEM shall be demonstrated by some selected investigations of WC-CO hardmeta. The results were obtained in collaboration by both groups with the aid of the pulsed CO2-laser. The TEA CO2 laser was transmitted through a ZnSe-window into the sample chamber of the SEM and focused ofAo tfte sample surface. It was operated in TEM - oo mode with a repetition rate of about 1 pulse per second. A peak power density of about 160 MW/cm2 was achieved in front of the sample surface.

  15. EVALUATION OF CONFOCAL MICROSCOPY SYSTEM PERFORMANCE

    EPA Science Inventory

    BACKGROUND. The confocal laser scanning microscope (CLSM) has enormous potential in many biological fields. Currently there is a subjective nature in the assessment of a confocal microscope's performance by primarily evaluating the system with a specific test slide provided by ea...

  16. Automatic analysis for neuron by confocal laser scanning microscope

    NASA Astrophysics Data System (ADS)

    Satou, Kouhei; Aoki, Yoshimitsu; Mataga, Nobuko; Hensh, Takao K.; Taki, Katuhiko

    2005-12-01

    The aim of this study is to develop a system that recognizes both the macro- and microscopic configurations of nerve cells and automatically performs the necessary 3-D measurements and functional classification of spines. The acquisition of 3-D images of cranial nerves has been enabled by the use of a confocal laser scanning microscope, although the highly accurate 3-D measurements of the microscopic structures of cranial nerves and their classification based on their configurations have not yet been accomplished. In this study, in order to obtain highly accurate measurements of the microscopic structures of cranial nerves, existing positions of spines were predicted by the 2-D image processing of tomographic images. Next, based on the positions that were predicted on the 2-D images, the positions and configurations of the spines were determined more accurately by 3-D image processing of the volume data. We report the successful construction of an automatic analysis system that uses a coarse-to-fine technique to analyze the microscopic structures of cranial nerves with high speed and accuracy by combining 2-D and 3-D image analyses.

  17. Modular Scanning Confocal Microscope with Digital Image Processing.

    PubMed

    Ye, Xianjun; McCluskey, Matthew D

    2016-01-01

    In conventional confocal microscopy, a physical pinhole is placed at the image plane prior to the detector to limit the observation volume. In this work, we present a modular design of a scanning confocal microscope which uses a CCD camera to replace the physical pinhole for materials science applications. Experimental scans were performed on a microscope resolution target, a semiconductor chip carrier, and a piece of etched silicon wafer. The data collected by the CCD were processed to yield images of the specimen. By selecting effective pixels in the recorded CCD images, a virtual pinhole is created. By analyzing the image moments of the imaging data, a lateral resolution enhancement is achieved by using a 20 × / NA = 0.4 microscope objective at 532 nm laser wavelength.

  18. Versatile optical system for static and dynamic thermomagnetic recording using a scanning laser microscope

    NASA Astrophysics Data System (ADS)

    Clegg, Warwick W.; Jenkins, David F. L.; Helian, Na; Windmill, James; Windmill, Robert

    2001-12-01

    Scanning Laser Microscopes (SLM) have been used to characterise the magnetic domain properties of various magnetic and magneto-optical materials. The SLM in our laboratory has been designed to enable both static and dynamic read-write operations to be performed on stationary media. In a conventional (static) SLM, data bits are recorded thermo-magnetically by focusing a pulse of laser light onto the sample surface. If the laser beam has a Gaussian intensity distribution (TEM00) then so will the focused laser spot. The resultant temperature profile will largely mirror the intensity distribution of the focused spot, and in the region where the temperature is sufficiently high for switching to occur, in the presence of bias field, a circular data bit will be recorded. However, in a real magneto-optical drive the bits are written onto non-stationary media, and the resultant bit will be non-circular. A versatile optical system has been developed to facilitate both recording and imaging of data bits. To simulate the action of a Magneto-Optical drive, the laser is pulsed via an Acousto-Optic Modulator, whilst being scanned across the sample using a galvanometer mounted mirror, thus imitating a storage medium rotating above a MO head with high relative velocity between the beam and medium. Static recording is simply achieved by disabling the galvanometer scan mirror. Polar magneto-optic Kerr effect images are acquired using multiple-segment photo-detectors for diffraction-limited scanned spot detection, with either specimen scanning for highest resolution or beam scanning for near real-time image acquisition. Results will be presented to illustrate the systems capabilities.

  19. Nanoscale welding of multi-walled carbon nanotubes by 1064 nm fiber laser

    NASA Astrophysics Data System (ADS)

    Yuan, Yanping; Liu, Zhi; Zhang, Kaihu; Han, Weina; Chen, Jimin

    2018-07-01

    This study proposes an efficient approach which uses 1064 nm continuous fiber laser to achieve nanoscale welding of crossed multi-walled carbon nanotubes (MWCNTs). By changing the irradiation time, different quality of nanoscale welding is obtained. The morphology changes are investigated by scanning electron microscope (SEM) and transmission electron microscope (TEM). The experiments demonstrate that better quality of MWCNTs nanoscale welding after 3 s irradiation can be obtained. It is found that new graphene layers between crossed nanotubes induced by laser make the nanoscale welding achieved due to the absorption of laser energy.

  20. Effect of scanning velocity on femtosecond laser-induced periodic surface structures on HgCdTe crystal

    NASA Astrophysics Data System (ADS)

    Gu, Hongan; Dai, Ye; Wang, Haodong; Yan, Xiaona; Ma, Guohong

    2017-12-01

    In this paper, a femtosecond laser line-scanning irradiation was used to induce the periodic surface microstructure on HgCdTe crystal. Low spatial frequency laser induced periodic surface structures of 650-770 nm and high spatial frequency laser induced periodic surface structures of 152-246 nm were respectively found with different scanning speeds. The evolution process from low spatial frequency laser induced periodic surface structures to high spatial frequency laser induced periodic surface structures is characterized by scanning electron microscope. Their spatial periods deduced by using a two-dimensional Fourier transformation partly agree with the predictions of the Sipe-Drude theory. Confocal micro-Raman spectral show that the atomic arrangement of induced low spatial frequency laser-induced structures are basically consistent with the crystal in the central area of laser-scanning line, however a new peak at 164 cm-1 for the CdTe-like mode becomes evident due to the Hg vaporization when strong laser ablation happens. The obtained surface periodic ripples may have applications in fabricating advanced infrared detector.

  1. Acquisition of multiple image stacks with a confocal laser scanning microscope

    NASA Astrophysics Data System (ADS)

    Zuschratter, Werner; Steffen, Thomas; Braun, Katharina; Herzog, Andreas; Michaelis, Bernd; Scheich, Henning

    1998-06-01

    Image acquisition at high magnification is inevitably correlated with a limited view over the entire tissue section. To overcome this limitation we designed software for multiple image-stack acquisition (3D-MISA) in confocal laser scanning microscopy (CLSM). The system consists of a 4 channel Leica CLSM equipped with a high resolution z- scanning stage mounted on a xy-monitorized stage. The 3D- MISA software is implemented into the microscope scanning software and uses the microscope settings for the movements of the xy-stage. It allows storage and recall of 70 xyz- positions and the automatic 3D-scanning of image arrays between selected xyz-coordinates. The number of images within one array is limited only by the amount of disk space or memory available. Although for most applications the accuracy of the xy-scanning stage is sufficient for a precise alignment of tiled views, the software provides the possibility of an adjustable overlap between two image stacks by shifting the moving steps of the xy-scanning stage. After scanning a tiled image gallery of the extended focus-images of each channel will be displayed on a graphic monitor. In addition, a tiled image gallery of individual focal planes can be created. In summary, the 3D-MISA allows 3D-image acquisition of coherent regions in combination with high resolution of single images.

  2. Surface wettability of silicon substrates enhanced by laser ablation

    NASA Astrophysics Data System (ADS)

    Tseng, Shih-Feng; Hsiao, Wen-Tse; Chen, Ming-Fei; Huang, Kuo-Cheng; Hsiao, Sheng-Yi; Lin, Yung-Sheng; Chou, Chang-Pin

    2010-11-01

    Laser-ablation techniques have been widely applied for removing material from a solid surface using a laser-beam irradiating apparatus. This paper presents a surface-texturing technique to create rough patterns on a silicon substrate using a pulsed Nd:YAG laser system. The different degrees of microstructure and surface roughness were adjusted by the laser fluence and laser pulse duration. A scanning electron microscope (SEM) and a 3D confocal laser-scanning microscope are used to measure the surface micrograph and roughness of the patterns, respectively. The contact angle variations between droplets on the textured surface were measured using an FTA 188 video contact angle analyzer. The results indicate that increasing the values of laser fluence and laser pulse duration pushes more molten slag piled around these patterns to create micro-sized craters and leads to an increase in the crater height and surface roughness. A typical example of a droplet on a laser-textured surface shows that the droplet spreads very quickly and almost disappears within 0.5167 s, compared to a contact angle of 47.9° on an untextured surface. This processing technique can also be applied to fabricating Si solar panels to increase the absorption efficiency of light.

  3. Laser scatter in clinical applications

    NASA Astrophysics Data System (ADS)

    Luther, Ed; Geddie, William

    2008-02-01

    Brightfield Laser Scanning Imaging (BLSI) is available on Laser Scanning Cytometers (LSCs) from CompuCyte Corporation. Briefly, digitation of photodetector outputs is coordinated with the combined motions of a small diameter (typically 2 to 10 microns) laser beam scanning a specimen in the Y direction (directed by a galvanometer-driven scanning mirror) and the microscope stage motion in the X direction. The output measurements are assembled into a two-dimensional array to provide a "non-real" digital image, where each pixel value reports the amount of laser-scattered light that is obtained when the laser beam is centered on that location. Depending on the detector positions, these images are analogous to Differential Interference Contrast or Phase Contrast microscopy. We report the incorporation of the new laser scattering capabilities into the workflow of a high-volume clinical cytology laboratory at University Health Network, Toronto, Canada. The laboratory has been employing LSC technology since 2003 for immunophenotypic fluorescence analysis of approximately 1200 cytological specimens per year, using the Clatch methodology. The new BLSI component allows visualization of cellular morphology at higher resolution levels than is possible with standard brightfield microscopic evaluation of unstained cells. BLSI is incorporated into the triage phase, where evaluation of unstained samples is combined with fluorescence evaluation to obtain specimen background levels. Technical details of the imaging methodology will be presented, as well as illustrative examples from current studies and comparisons to detailed, but obscure, historical studies of cytology specimens based on phase contrast microscopy.

  4. How the confocal laser scanning microscope entered biological research.

    PubMed

    Amos, W B; White, J G

    2003-09-01

    A history of the early development of the confocal laser scanning microscope in the MRC Laboratory of Molecular Biology in Cambridge is presented. The rapid uptake of this technology is explained by the wide use of fluorescence in the 80s. The key innovations were the scanning of the light beam over the specimen rather than vice-versa and a high magnification at the level of the detector, allowing the use of a macroscopic iris. These were followed by an achromatic all-reflective relay system, a non-confocal transmission detector and novel software for control and basic image processing. This design was commercialized successfully and has been produced and developed over 17 years, surviving challenges from alternative technologies, including solid-state scanning systems. Lessons are pointed out from the unusual nature of the original funding and research environment. Attention is drawn to the slow adoption of the instrument in diagnostic medicine, despite promising applications.

  5. Three-dimensional scanning confocal laser microscope

    DOEpatents

    Anderson, R. Rox; Webb, Robert H.; Rajadhyaksha, Milind

    1999-01-01

    A confocal microscope for generating an image of a sample includes a first scanning element for scanning a light beam along a first axis, and a second scanning element for scanning the light beam at a predetermined amplitude along a second axis perpendicular to the first axis. A third scanning element scans the light beam at a predetermined amplitude along a third axis perpendicular to an imaging plane defined by the first and second axes. The second and third scanning element are synchronized to scan at the same frequency. The second and third predetermined amplitudes are percentages of their maximum amplitudes. A selector determines the second and third predetermined amplitudes such that the sum of the percentages is equal to one-hundred percent.

  6. Automated Image Analysis Corrosion Working Group Update: February 1, 2018

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

    Wendelberger, James G.

    These are slides for the automated image analysis corrosion working group update. The overall goals were: automate the detection and quantification of features in images (faster, more accurate), how to do this (obtain data, analyze data), focus on Laser Scanning Confocal Microscope (LCM) data (laser intensity, laser height/depth, optical RGB, optical plus laser RGB).

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

    PubMed Central

    Negrean, Adrian; Mansvelder, Huibert D.

    2014-01-01

    In laser-scanning microscopy often an off-the-shelf achromatic doublet is used as a scan lens which can reduce the available diffraction-limited field-of-view (FOV) by a factor of 3 and introduce chromatic aberrations that are scan angle dependent. Here we present several simple lens designs of superior quality that fully make use of high-NA low-magnification objectives, offering diffraction-limited imaging over a large FOV and wavelength range. We constructed a two-photon laser-scanning microscope with optimized custom lenses which had a near diffraction limit point-spread-function (PSF) with less than 3.6% variation over a 400 µm FOV and less than 0.5 µm lateral color between 750 and 1050 nm. PMID:24877017

  8. Microscope and method of use

    DOEpatents

    Bongianni, Wayne L.

    1984-01-01

    A method and apparatus for electronically focusing and electronically scanning microscopic specimens are given. In the invention, visual images of even moving, living, opaque specimens can be acoustically obtained and viewed with virtually no time needed for processing (i.e., real time processing is used). And planar samples are not required. The specimens (if planar) need not be moved during scanning, although it will be desirable and possible to move or rotate nonplanar specimens (e.g., laser fusion targets) against the lens of the apparatus. No coupling fluid is needed, so specimens need not be wetted. A phase acoustic microscope is also made from the basic microscope components together with electronic mixers.

  9. Microscope and method of use

    DOEpatents

    Bongianni, W.L.

    1984-04-17

    A method and apparatus for electronically focusing and electronically scanning microscopic specimens are given. In the invention, visual images of even moving, living, opaque specimens can be acoustically obtained and viewed with virtually no time needed for processing (i.e., real time processing is used). And planar samples are not required. The specimens (if planar) need not be moved during scanning, although it will be desirable and possible to move or rotate nonplanar specimens (e.g., laser fusion targets) against the lens of the apparatus. No coupling fluid is needed, so specimens need not be wetted. A phase acoustic microscope is also made from the basic microscope components together with electronic mixers. 7 figs.

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

    PubMed

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

    2003-05-01

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

  11. Application of laser scanning confocal microscopy in the soft tissue exquisite structure for 3D scan

    PubMed Central

    Zhang, Zhaoqiang; Ibrahim, Mohamed; Fu, Yang; Wu, Xujia; Ren, Fei; Chen, Lei

    2018-01-01

    Three-dimensional (3D) printing is a new developing technology for printing individualized materials swiftly and precisely in the field of biological medicine (especially tissue-engineered materials). Prior to printing, it is necessary to scan the structure of the natural biological tissue, then construct the 3D printing digital model through optimizing the scanned data. By searching the literatures, magazines at home and abroad, this article reviewed the current status, main processes and matters needing attention of confocal laser scanning microscope (LSCM) in the application of soft tissue fine structure 3D scanning, empathizing the significance of LSCM in this field. PMID:29755838

  12. Modular Scanning Confocal Microscope with Digital Image Processing

    PubMed Central

    McCluskey, Matthew D.

    2016-01-01

    In conventional confocal microscopy, a physical pinhole is placed at the image plane prior to the detector to limit the observation volume. In this work, we present a modular design of a scanning confocal microscope which uses a CCD camera to replace the physical pinhole for materials science applications. Experimental scans were performed on a microscope resolution target, a semiconductor chip carrier, and a piece of etched silicon wafer. The data collected by the CCD were processed to yield images of the specimen. By selecting effective pixels in the recorded CCD images, a virtual pinhole is created. By analyzing the image moments of the imaging data, a lateral resolution enhancement is achieved by using a 20 × / NA = 0.4 microscope objective at 532 nm laser wavelength. PMID:27829052

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

    NASA Technical Reports Server (NTRS)

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

    1981-01-01

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

  14. Femtosecond laser for cavity preparation in enamel and dentin: ablation efficiency related factors.

    PubMed

    Chen, H; Li, H; Sun, Yc; Wang, Y; Lü, Pj

    2016-02-11

    To study the effects of laser fluence (laser energy density), scanning line spacing and ablation depth on the efficiency of a femtosecond laser for three-dimensional ablation of enamel and dentin. A diode-pumped, thin-disk femtosecond laser (wavelength 1025 nm, pulse width 400 fs) was used for the ablation of enamel and dentin. The laser spot was guided in a series of overlapping parallel lines on enamel and dentin surfaces to form a three-dimensional cavity. The depth and volume of the ablated cavity was then measured under a 3D measurement microscope to determine the ablation efficiency. Different values of fluence, scanning line spacing and ablation depth were used to assess the effects of each variable on ablation efficiency. Ablation efficiencies for enamel and dentin were maximized at different laser fluences and number of scanning lines and decreased with increases in laser fluence or with increases in scanning line spacing beyond spot diameter or with increases in ablation depth. Laser fluence, scanning line spacing and ablation depth all significantly affected femtosecond laser ablation efficiency. Use of a reasonable control for each of these parameters will improve future clinical application.

  15. Effect of the Femtosecond Laser on an Intracorneal Inlay for Surgical Compensation of Presbyopia during Cataract Surgery: Scanning Electron Microscope Imaging.

    PubMed

    Ibarz, Marta; Rodríguez-Prats, Jose Luis; Hernández-Verdejo, Jose Luis; Tañá, Pedro

    2017-02-01

    To investigate the effect of the femtosecond laser-assisted cataract surgery (FLACS) on porcine eyes implanted with a Kamra corneal inlay and to describe how the inlay may change the effect of the femtosecond laser on the lens. FLACS was performed on six porcine eyes and a Kamra corneal inlay had been implanted, exploring the lens under the surgical microscope. Another Kamra corneal inlay was attached to the upper part of the transparent hemisphere used for calibration of the femtosecond laser. Capsulorhexis, arcuate incisions, and phacofragmentation were carried out. The Kamra corneal inlay was compared with a nontreated one using a scanning electron microscope (SEM), and the hemisphere was analyzed with a surgical microscope. Capsulorhexis and phacofragmentation were completed in all the porcine eyes, although accuracy to determine the exact effect on the lens was not possible to achieve. The effect of the femtosecond laser on the PMMA hemisphere through the Kamra corneal inlay showed the capsulorhexis was placed outside the outer margin of the inlay and a sharply sculpted fragmentation pattern with a three-dimensional (donut-shaped) annulus untreated beneath it. SEM images of the nontreated and the treated inlays were comparable. No ultrastructural changes were found in the treated Kamra corneal inlay. FLACS can be performed with a Kamra corneal inlay for surgical compensation of presbyopia without the risk of damaging the inlay. The Kamra corneal inlay acts as a screen that avoids the laser to reach the areas beneath its shadow, but not the exposed areas of the lens.

  16. Two-Photon Fluorescence Microscope for Microgravity Research

    NASA Technical Reports Server (NTRS)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2005-01-01

    A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the longer-wavelength excitation light and passes the shorter-wavelength fluorescence light. Also, the confocal pinhole has been removed to increase the signal strength. The laser beam is scanned by a twoperpendicular- axis pair of galvanometer mirrors through a pupil transfer lens into the side port of an inverted microscope. Finally, the beam is focused by a 63-magnification, 1.3-numerical- aperture oil-immersion objective lens onto a specimen. The pupil transfer lens serves to match the intermediate image planes of the scanning head and the microscope, and its location is critical. In order to maximize the quality of the image, (that is, the point spread function of the objective lens for all scan positions), the entire system was modeled in optical-design software, and the various free design parameters (the parameters of the spatial-filter components as well as the separations of all of the system components) were determined through an iterative optimization process. A modular design was chosen to facilitate access to the optical train for future fluorescence correlation spectroscopy and fluorescence-lifetime experiments.

  17. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: SPECTROSCOPY AND FOUNDATIONS FOR QUANTITATION

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The reliability of the CLSM to obtain specific measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. For man...

  18. Demonstration of a plenoptic microscope based on laser optical feedback imaging.

    PubMed

    Glastre, Wilfried; Hugon, Olivier; Jacquin, Olivier; Guillet de Chatellus, Hugues; Lacot, Eric

    2013-03-25

    A new kind of plenoptic imaging system based on Laser Optical Feedback Imaging (LOFI) is presented and is compared to another previously existing device based on microlens array. Improved photometric performances, resolution and depth of field are obtained at the price of a slow point by point scanning. Main properties of plenoptic microscopes such as numerical refocusing on any curved surface or aberrations compensation are both theoretically and experimentally demonstrated with a LOFI-based device.

  19. Development of a New Punch Head Shape to Replicate Scale-Up Issues on a Laboratory Tablet Press III: Replicating sticking phenomenon using the SAS punch and evaluation by checking the tablet surface using 3D laser scanning microscope.

    PubMed

    Ito, Manabu; Aoki, Shigeru; Uchiyama, Jumpei; Yamato, Keisuke

    2018-04-20

    Sticking is a common observation in the scale-up stage on the punch tip using a commercial tableting machine. The difference in the total compression time between a laboratory and a commercial tableting machine is considered one of the main root causes of scale up issues in the tableting processes. The proposed Size Adjusted for Scale-up (SAS) punch can be used to adjust the consolidation and dwell times for commercial tableting machine. As a result, the sticking phenomenon is able to be replicated at the pilot scale stage. As reported in this paper, the quantification of sticking was measured using a 3D laser scanning microscope to check the tablet surface. It was shown that the sticking area decreased with the addition of magnesium stearate in the formulation, but the sticking depth was not affected by the additional amount of magnesium stearate. It is proposed that use of a 3D laser scanning microscope can be applied to evaluate sticking as a process analytical technology (PAT) tool and so sticking can be monitored continuously without stopping the machine. Copyright © 2018. Published by Elsevier Inc.

  20. Multimodal nonlinear microscope based on a compact fiber-format laser source

    NASA Astrophysics Data System (ADS)

    Crisafi, Francesco; Kumar, Vikas; Perri, Antonio; Marangoni, Marco; Cerullo, Giulio; Polli, Dario

    2018-01-01

    We present a multimodal non-linear optical (NLO) laser-scanning microscope, based on a compact fiber-format excitation laser and integrating coherent anti-Stokes Raman scattering (CARS), stimulated Raman scattering (SRS) and two-photon-excitation fluorescence (TPEF) on a single platform. We demonstrate its capabilities in simultaneously acquiring CARS and SRS images of a blend of 6-μm poly(methyl methacrylate) beads and 3-μm polystyrene beads. We then apply it to visualize cell walls and chloroplast of an unprocessed fresh leaf of Elodea aquatic plant via SRS and TPEF modalities, respectively. The presented NLO microscope, developed in house using off-the-shelf components, offers full accessibility to the optical path and ensures its easy re-configurability and flexibility.

  1. Visible-to-visible four-photon ultrahigh resolution microscopic imaging with 730-nm diode laser excited nanocrystals.

    PubMed

    Wang, Baoju; Zhan, Qiuqiang; Zhao, Yuxiang; Wu, Ruitao; Liu, Jing; He, Sailing

    2016-01-25

    Further development of multiphoton microscopic imaging is confronted with a number of limitations, including high-cost, high complexity and relatively low spatial resolution due to the long excitation wavelength. To overcome these problems, for the first time, we propose visible-to-visible four-photon ultrahigh resolution microscopic imaging by using a common cost-effective 730-nm laser diode to excite the prepared Nd(3+)-sensitized upconversion nanoparticles (Nd(3+)-UCNPs). An ordinary multiphoton scanning microscope system was built using a visible CW diode laser and the lateral imaging resolution as high as 161-nm was achieved via the four-photon upconversion process. The demonstrated large saturation excitation power for Nd(3+)-UCNPs would be more practical and facilitate the four-photon imaging in the application. A sample with fine structure was imaged to demonstrate the advantages of visible-to-visible four-photon ultrahigh resolution microscopic imaging with 730-nm diode laser excited nanocrystals. Combining the uniqueness of UCNPs, the proposed visible-to-visible four-photon imaging would be highly promising and attractive in the field of multiphoton imaging.

  2. Laser excited confocal microscope fluorescence scanner and method

    DOEpatents

    Mathies, Richard A.; Peck, Konan

    1992-01-01

    A fluorescent scanner for scanning the fluorescence from a fluorescence labeled separated sample on a sample carrier including a confocal microscope for illuminating a predetermined volume of the sample carrier and/or receiving and processing fluorescence emissions from said volume to provide a display of the separated sample.

  3. In Situ Observation of Kinetic Processes of Lath Bainite Nucleation and Growth by Laser Scanning Confocal Microscope in Reheated Weld Metals

    NASA Astrophysics Data System (ADS)

    Mao, Gaojun; Cao, Rui; Guo, Xili; Jiang, Yong; Chen, Jianhong

    2017-12-01

    The kinetic processes of nucleation and growth of bainite laths in reheated weld metals are observed and analyzed by a combination of a laser confocal scanning microscope and an electron backscattering diffraction with a field emission scanning electron microscope. The results indicate that the surface relief induced by phase transformation is able to reveal the real microstructural morphologies of bainite laths when viewed from various angles. Five nucleation modes and six types of growth behaviors of bainite laths are revealed. The bainite lath growth rates are measured to vary over a wide range, from 2 μm/s to higher than 2000 μm/s. The orientations of the bainite laths within a prior austenite grain are examined and denoted as different variants. On the basis of variant identification, the reason is analyzed for various growth rates which are demonstrated to be affected by (1) the density of the high-angle misorientation in it, (2) the included angle between habit planes of different variants, and (3) the direction of lath growth with respect to the free (polished) surface.

  4. Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons.

    PubMed Central

    Fan, G Y; Fujisaki, H; Miyawaki, A; Tsay, R K; Tsien, R Y; Ellisman, M H

    1999-01-01

    A video-rate (30 frames/s) scanning two-photon excitation microscope has been successfully tested. The microscope, based on a Nikon RCM 8000, incorporates a femtosecond pulsed laser with wavelength tunable from 690 to 1050 nm, prechirper optics for laser pulse-width compression, resonant galvanometer for video-rate point scanning, and a pair of nonconfocal detectors for fast emission ratioing. An increase in fluorescent emission of 1.75-fold is consistently obtained with the use of the prechirper optics. The nonconfocal detectors provide another 2.25-fold increase in detection efficiency. Ratio imaging and optical sectioning can therefore be performed more efficiently without confocal optics. Faster frame rates, at 60, 120, and 240 frames/s, can be achieved with proportionally reduced scan lines per frame. Useful two-photon images can be acquired at video rate with a laser power as low as 2.7 mW at specimen with the genetically modified green fluorescent proteins. Preliminary results obtained using this system confirm that the yellow "cameleons" exhibit similar optical properties as under one-photon excitation conditions. Dynamic two-photon images of cardiac myocytes and ratio images of yellow cameleon-2.1, -3.1, and -3.1nu are also presented. PMID:10233058

  5. High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

    PubMed

    Bullen, A; Patel, S S; Saggau, P

    1997-07-01

    The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.

  6. High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

    PubMed Central

    Bullen, A; Patel, S S; Saggau, P

    1997-01-01

    The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging. Images FIGURE 6 PMID:9199810

  7. Laser-induced selective metallization of polypropylene doped with multiwall carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Ratautas, Karolis; Gedvilas, Mindaugas; Stankevičiene, Ina; Jagminienė, Aldona; Norkus, Eugenijus; Pira, Nello Li; Sinopoli, Stefano; Račiukaitis, Gediminas

    2017-08-01

    Moulded interconnect devices (MID) offer the material, weight and cost saving by integration electronic circuits directly into polymeric components used in automotive and other consumer products. Lasers are used to write circuits directly by modifying the surface of polymers followed by an electroless metal plating. A new composite material - the polypropylene doped with multiwall carbon nanotubes was developed for the laser-induced selective metallization. Mechanism of surface activation by laser irradiation was investigated in details utilising pico- and nanoseconds lasers. Deposition of copper was performed in the autocatalytic electroless plating bath. The laser-activated polymer surfaces have been studied using the Raman spectroscopy and scanning electron microscope (SEM). Microscopic images revealed that surface becomes active only after its melting by a laser. Alterations in the Raman spectra of the D and G bands indicated the clustering of carbon additives in the composite material. Optimal laser parameters for the surface activation were found by measuring a sheet resistance of the finally metal-plated samples. A spatially selective copper plating was achieved with the smallest conductor line width of 22 μm at the laser scanning speed of 3 m/s and the pulse repetition rate of 100 kHz. Finally, the technique was validated by making functional electronic circuits by this MID approach.

  8. Demonstration of bacterial biofilms in culture-negative silicone stent and jones tube.

    PubMed

    Parsa, Kami; Schaudinn, Christoph; Gorur, Amita; Sedghizadeh, Parish P; Johnson, Thomas; Tse, David T; Costerton, John W

    2010-01-01

    To demonstrate the presence of bacterial biofilms on a dacryocystorhinostomy silicone stent and a Jones tube. One dacryocystorhinostomy silicone stent and one Jones tube were removed from 2 patients who presented with an infection of their respective nasolacrimal system. Cultures were obtained, and the implants were processed for scanning electron microscopy and confocal laser scanning microscopy, advanced microscopic methods that are applicable for detection of uncultivable biofilm organisms. Routine bacterial cultures revealed no growth, but bacterial biofilms on outer and inner surfaces of both implants were confirmed by advanced microscopic techniques. To the authors' knowledge, this is the first article that documents the presence of biofilms on a Crawford stent or a Jones tube on patients who presented with infections involving the nasolacrimal system. Although initial cultures revealed absence of any bacterial growth, confocal laser scanning microscopy and scanning electron microscopy documented bacterial colonization. Clinicians should consider the role of biofilms and the limitation of our standard culturing techniques while treating patients with device- or implant-related infections.

  9. Seamless stitching of tile scan microscope images.

    PubMed

    Legesse, F B; Chernavskaia, O; Heuke, S; Bocklitz, T; Meyer, T; Popp, J; Heintzmann, R

    2015-06-01

    For diagnostic purposes, optical imaging techniques need to obtain high-resolution images of extended biological specimens in reasonable time. The field of view of an objective lens, however, is often smaller than the sample size. To image the whole sample, laser scanning microscopes acquire tile scans that are stitched into larger mosaics. The appearance of such image mosaics is affected by visible edge artefacts that arise from various optical aberrations which manifest in grey level jumps across tile boundaries. In this contribution, a technique for stitching tiles into a seamless mosaic is presented. The stitching algorithm operates by equilibrating neighbouring edges and forcing the brightness at corners to a common value. The corrected image mosaics appear to be free from stitching artefacts and are, therefore, suited for further image analysis procedures. The contribution presents a novel method to seamlessly stitch tiles captured by a laser scanning microscope into a large mosaic. The motivation for the work is the failure of currently existing methods for stitching nonlinear, multimodal images captured by our microscopic setups. Our method eliminates the visible edge artefacts that appear between neighbouring tiles by taking into account the overall illumination differences among tiles in such mosaics. The algorithm first corrects the nonuniform brightness that exists within each of the tiles. It then compensates for grey level differences across tile boundaries by equilibrating neighbouring edges and forcing the brightness at the corners to a common value. After these artefacts have been removed further image analysis procedures can be applied on the microscopic images. Even though the solution presented here is tailored for the aforementioned specific case, it could be easily adapted to other contexts where image tiles are assembled into mosaics such as in astronomical or satellite photos. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  10. Influence of Ultrasonic Surface Rolling on Microstructure and Wear Behavior of Selective Laser Melted Ti-6Al-4V Alloy

    PubMed Central

    Wang, Zhen; Xiao, Zhiyu; Huang, Chuanshou; Wen, Liping; Zhang, Weiwen

    2017-01-01

    The present article studied the effect of ultrasonic surface rolling process (USRP) on the microstructure and wear behavior of a selective laser melted Ti-6Al-4V alloy. Surface characteristics were investigated using optical microscope, nano-indentation, scanning electron microscope, transmission electron microscope and laser scanning confocal microscope. Results indicated that the thickness of pore-free surfaces increased to 100~200 μm with the increasing ultrasonic surface rolling numbers. Severe work hardening occurred in the densified layer, resulting in the formation of refined grains, dislocation walls and deformation twins. After 1000 N 6 passes, about 15.5% and 14.1% increment in surficial Nano-hardness and Vickers-hardness was obtained, respectively. The hardness decreased gradually from the top surface to the substrate. Wear tests revealed that the friction coefficient declined from 0.74 (polished surface) to 0.64 (USRP treated surface) and the wear volume reduced from 0.205 mm−3 to 0.195 mm−3. The difference in wear volume between USRP treated and polished samples increased with sliding time. The enhanced wear resistance was concluded to be associated with the improvement of hardness and shear resistance and also the inhibition of delamination initiation. PMID:29048344

  11. Laser shock wave assisted patterning on NiTi shape memory alloy surfaces

    NASA Astrophysics Data System (ADS)

    Seyitliyev, Dovletgeldi; Li, Peizhen; Kholikov, Khomidkhodza; Grant, Byron; Karaca, Haluk E.; Er, Ali O.

    2017-02-01

    An advanced direct imprinting method with low cost, quick, and less environmental impact to create thermally controllable surface pattern using the laser pulses is reported. Patterned micro indents were generated on Ni50Ti50 shape memory alloys (SMA) using an Nd:YAG laser operating at 1064 nm combined with suitable transparent overlay, a sacrificial layer of graphite, and copper grid. Laser pulses at different energy densities which generates pressure pulses up to 10 GPa on the surface was focused through the confinement medium, ablating the copper grid to create plasma and transferring the grid pattern onto the NiTi surface. Scanning electron microscope (SEM) and optical microscope images of square pattern with different sizes were studied. One dimensional profile analysis shows that the depth of the patterned sample initially increase linearly with the laser energy until 125 mJ/pulse where the plasma further absorbs and reflects the laser beam. In addition, light the microscope image show that the surface of NiTi alloy was damaged due to the high power laser energy which removes the graphite layer.

  12. Laser excited confocal microscope fluorescence scanner and method

    DOEpatents

    Mathies, R.A.; Peck, K.

    1992-02-25

    A fluorescent scanner is designed for scanning the fluorescence from a fluorescence labeled separated sample on a sample carrier. The scanner includes a confocal microscope for illuminating a predetermined volume of the sample carrier and/or receiving and processing fluorescence emissions from the volume to provide a display of the separated sample. 8 figs.

  13. Development and applications of optical interferometric micrometrology in the Angstrom and subangstrom range

    NASA Technical Reports Server (NTRS)

    Lauer, James L.; Abel, Phillip B.

    1988-01-01

    The characteristics of the scanning tunneling microscope and atomic force microscope (AFM) are briefly reviewed, and optical methods, mainly interferometry, of sufficient resolution to measure AFM deflections are discussed. The methods include optical resonators, laser interferometry, multiple-beam interferometry, and evanescent wave detection. Experimental results using AFM are reviewed.

  14. Demodulation signal processing in multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Fisher, Walter G.; Wachter, Eric A.; Piston, David W.

    2002-06-01

    Multiphoton laser scanning microscopy offers numerous advantages, but sensitivity can be seriously affected by contamination from ambient room light. Typically, this forces experiments to be performed in an absolutely dark room. Since mode-locked lasers are used to generate detectable signals, signal-processing can be used to avoid such problems by taking advantage of the pulsed characteristics of such lasers. Demodulation of the fluorescence signal generated at the mode-locked frequency can result in significant reduction of interference from ambient noise sources. Such demodulation can be readily adapted to existing microscopes by inserting appropriate processor circuitry between the detector and data collection system, yielding a more robust microscope.

  15. High temporal and spatial resolution studies of bone cells using real-time confocal reflection microscopy.

    PubMed

    Boyde, A; Vesely, P; Gray, C; Jones, S J

    1994-01-01

    Chick and rat bone-derived cells were mounted in sealed coverslip-covered chambers; individual osteoclasts (but also osteoblasts) were selected and studied at 37 degrees C using three different types of high-speed scanning confocal microscopes: (1) A Noran Tandem Scanning Microscope (TSM) was used with a low light level, cooled CCD camera for image transfer to a Noran TN8502 frame store-based image analysing computer to make time lapse movie sequences using 0.1 s exposure periods, thus losing some of the advantage of the high frame rate of the TSM. Rapid focus adjustment using computer controlled piezo drivers permitted two or more focus planes to be imaged sequentially: thus (with additional light-source shuttering) the reflection confocal image could be alternated with the phase contrast image at a different focus. Individual cells were followed for up to 5 days, suggesting no significant irradiation problem. (2) Exceptional temporal and spatial resolution is available in video rate laser confocal scanning microscopes (VRCSLMs). We used the Noran Odyssey unitary beam VRCSLM with an argon ion laser at 488 nm and acousto-optic deflection (AOD) on the line axis: this instrument is truly and adjustably confocal in the reflection mode. (3) We also used the Lasertec 1LM11 line scan instrument, with an He-Ne laser at 633 nm, and AOD for the frame scan. We discuss the technical problems and merits of the different approaches. The VRCSLMs documented rapid, real-time oscillatory motion: all the methods used show rapid net movement of organelles within bone cells. The interference reflection mode gives particularly strong contrasts in confocal instruments. Phase contrast and other interference methods used in the microscopy of living cells can be used simultaneously in the TSM.

  16. Evaluation of Enterococcus faecalis adhesion, penetration, and method to prevent the penetration of Enterococcus faecalis into root cementum: Confocal laser scanning microscope and scanning electron microscope analysis.

    PubMed

    Halkai, Rahul S; Hegde, Mithra N; Halkai, Kiran R

    2016-01-01

    To ascertain the role of Enterococcus faecalis in persistent infection and a possible method to prevent the penetration of E. faecalis into root cementum. One hundred and twenty human single-rooted extracted teeth divided into five groups. Group I (control): intact teeth, Group II: no apical treatment done, Group III divided into two subgroups. In Groups IIIa and IIIb, root apex treated with lactic acid of acidic and neutral pH, respectively. Group IV: apical root cementum exposed to lactic acid and roughened to mimic the apical resorption. Group V: apical treatment done same as Group IV and root-end filling done using mineral trioxide aggregate (MTA). Apical one-third of all samples immersed in E. faecalis broth for 8 weeks followed by bone morphogenetic protein and obturation and again immersed into broth for 8 weeks. Teeth split into two halves and observed under confocal laser scanning microscope and scanning electron microscope, organism identified by culture and polymerase chain reaction techniques. Adhesion and penetration was observed in Group IIIa and Group IV. Only adhesion in Group II and IIIB and no adhesion and penetration in Group I and V. Adhesion and penetration of E. faecalis into root cementum providing a long-term nidus for subsequent infection are the possible reason for persistent infection and root-end filling with MTA prevents the adhesion and penetration.

  17. Any Way You Slice It—A Comparison of Confocal Microscopy Techniques

    PubMed Central

    Jonkman, James

    2015-01-01

    The confocal fluorescence microscope has become a popular tool for life sciences researchers, primarily because of its ability to remove blur from outside of the focal plane of the image. Several different kinds of confocal microscopes have been developed, each with advantages and disadvantages. This article will cover the grid confocal, classic confocal laser-scanning microscope (CLSM), the resonant scanning-CLSM, and the spinning-disk confocal microscope. The way each microscope technique works, the best applications the technique is suited for, the limitations of the technique, and new developments for each technology will be presented. Researchers who have access to a range of different confocal microscopes (e.g., through a local core facility) should find this paper helpful for choosing the best confocal technology for specific imaging applications. Others with funding to purchase an instrument should find the article helpful in deciding which technology is ideal for their area of research. PMID:25802490

  18. Laser scanning confocal microscopy: history, applications, and related optical sectioning techniques.

    PubMed

    Paddock, Stephen W; Eliceiri, Kevin W

    2014-01-01

    Confocal microscopy is an established light microscopical technique for imaging fluorescently labeled specimens with significant three-dimensional structure. Applications of confocal microscopy in the biomedical sciences include the imaging of the spatial distribution of macromolecules in either fixed or living cells, the automated collection of 3D data, the imaging of multiple labeled specimens and the measurement of physiological events in living cells. The laser scanning confocal microscope continues to be chosen for most routine work although a number of instruments have been developed for more specific applications. Significant improvements have been made to all areas of the confocal approach, not only to the instruments themselves, but also to the protocols of specimen preparation, to the analysis, the display, the reproduction, sharing and management of confocal images using bioinformatics techniques.

  19. Diffraction-Unlimited Fluorescence Imaging with an EasySTED Retrofitted Confocal Microscope.

    PubMed

    Klauss, André; Hille, Carsten

    2017-01-01

    The easySTED technology provides the means to retrofit a confocal microscope to a diffraction-unlimited stimulated emission depletion (STED) microscope.Although commercial STED systems are available today, for many users of confocal laser scanning microscopes the option of retrofitting their confocal system to a STED system ready for diffraction-unlimited imaging may present an attractive option. The easySTED principle allowing for a joint beam path of excitation and depletion light promises some advantages concerning technical complexity and alignment effort for such an STED upgrade. In the one beam path design of easySTED the use of a common laser source, either a supercontinuum source or two separate lasers coupled into the same single-mode fiber, becomes feasible. The alignment of the focal light distribution of the STED beam relative to that of the excitation beam in all three spatial dimensions is therefore omitted respectively reduced to coupling the STED laser into the common single-mode fiber. Thus, only minor modifications need to be applied to the beam path in the confocal microscope to be upgraded. Those comprise adding polarization control elements and the easySTED waveplate, and adapting the beamsplitter to the excitation/STED wavelength combination.

  20. Apertureless near-field scanning optical microscope working with or without laser source.

    PubMed

    Formanek, F; De Wilde, Y; Aigouy, L; Chen, Y

    2004-01-01

    An apertureless near-field scanning optical microscope (ANSOM), used indifferent configurations, is presented. Our versatile home-made setup, based on a sharp tungsten tip glued onto a quartz tuning fork and working in tapping mode, allows to perform imaging over a broad spectral range. We have recorded optical images in the visible (wavelength, lambda = 655 nm) and in the infrared (lambda = 10.6 microm), proving that the setup routinely achieves an optical resolution of <50 nm regardless of the illumination wavelength. We have also shown optical images recorded in the visible (lambda = 655 nm) in an inverted configuration where the tip does not perturb the focused spot of the illumination laser. Approach curves as well as image profiles have revealed that on demodulating the optical signal at higher harmonics, we can obtain an effective probe sharpening which results in an improvement of the resolution. Finally, we have presented optical images recorded in the infrared without any illumination, that is, the usual laser source is replaced by a simple heating of the sample. This has shown that the ANSOM can be used as a near-field thermal optical microscope (NTOM) to probe the near field generated by the thermal emission of the sample.

  1. Imaging fluorescence detected linear dichroism of plant cell walls in laser scanning confocal microscope.

    PubMed

    Steinbach, Gábor; Pomozi, István; Zsiros, Ottó; Páy, Anikó; Horváth, Gábor V; Garab, Gyozo

    2008-03-01

    Anisotropy carries important information on the molecular organization of biological samples. Its determination requires a combination of microscopy and polarization spectroscopy tools. The authors constructed differential polarization (DP) attachments to a laser scanning microscope in order to determine physical quantities related to the anisotropic distribution of molecules in microscopic samples; here the authors focus on fluorescence-detected linear dichroism (FDLD). By modulating the linear polarization of the laser beam between two orthogonally polarized states and by using a demodulation circuit, the authors determine the associated transmitted and fluorescence intensity-difference signals, which serve the basis for LD (linear dichroism) and FDLD, respectively. The authors demonstrate on sections of Convallaria majalis root tissue stained with Acridin Orange that while (nonconfocal) LD images remain smeared and weak, FDLD images recorded in confocal mode reveal strong anisotropy of the cell wall. FDLD imaging is suitable for mapping the anisotropic distribution of transition dipoles in 3 dimensions. A mathematical model is proposed to account for the fiber-laminate ultrastructure of the cell wall and for the intercalation of the dye molecules in complex, highly anisotropic architecture. Copyright 2007 International Society for Analytical Cytology.

  2. Characterisation of a resolution enhancing image inversion interferometer.

    PubMed

    Wicker, Kai; Sindbert, Simon; Heintzmann, Rainer

    2009-08-31

    Image inversion interferometers have the potential to significantly enhance the lateral resolution and light efficiency of scanning fluorescence microscopes. Self-interference of a point source's coherent point spread function with its inverted copy leads to a reduction in the integrated signal for off-axis sources compared to sources on the inversion axis. This can be used to enhance the resolution in a confocal laser scanning microscope. We present a simple image inversion interferometer relying solely on reflections off planar surfaces. Measurements of the detection point spread function for several types of light sources confirm the predicted performance and suggest its usability for scanning confocal fluorescence microscopy.

  3. Two-photon imaging in living brain slices.

    PubMed

    Mainen, Z F; Maletic-Savatic, M; Shi, S H; Hayashi, Y; Malinow, R; Svoboda, K

    1999-06-01

    Two-photon excitation laser scanning microscopy (TPLSM) has become the tool of choice for high-resolution fluorescence imaging in intact neural tissues. Compared with other optical techniques, TPLSM allows high-resolution imaging and efficient detection of fluorescence signal with minimal photobleaching and phototoxicity. The advantages of TPLSM are especially pronounced in highly scattering environments such as the brain slice. Here we describe our approaches to imaging various aspects of synaptic function in living brain slices. To combine several imaging modes together with patch-clamp electrophysiological recordings we found it advantageous to custom-build an upright microscope. Our design goals were primarily experimental convenience and efficient collection of fluorescence. We describe our TPLSM imaging system and its performance in detail. We present dynamic measurements of neuronal morphology of neurons expressing green fluorescent protein (GFP) and GFP fusion proteins as well as functional imaging of calcium dynamics in individual dendritic spines. Although our microscope is a custom instrument, its key advantages can be easily implemented as a modification of commercial laser scanning microscopes. Copyright 1999 Academic Press.

  4. Quantification of incisal tooth wear in upper anterior teeth: conventional vs new method using toolmakers microscope and a three-dimensional measuring technique.

    PubMed

    Al-Omiri, Mahmoud K; Sghaireen, Mohd G; Alzarea, Bader K; Lynch, Edward

    2013-12-01

    This study aimed to quantify tooth wear in upper anterior teeth using a new CAD-CAM Laser scanning machine, tool maker microscope and conventional tooth wear index. Fifty participants (25 males and 25 females, mean age = 25 ± 4 years) were assessed for incisal tooth wear of upper anterior teeth using Smith and Knight clinical tooth wear index (TWI) on two occasions, the study baseline and 1 year later. Stone dies for each tooth were prepared and scanned using the CAD-CAM Laser Cercon System. Scanned images were printed and examined under a toolmaker microscope to quantify tooth wear and then the dies were directly assessed under the microscope to measure tooth wear. The Wilcoxon Signed Ranks Test was used to analyze the data. TWI scores for incisal edges were 0-3 and were similar at both occasions. Score 4 was not detected. Wear values measured by directly assessing the dies under the toolmaker microscope (range = 113 - 150 μm, mean = 130 ± 20 μm) were significantly more than those measured from Cercon Digital Machine images (range=52-80 μm, mean = 68 ± 23 μm) and both showed significant differences between the two occasions. Wear progression in upper anterior teeth was effectively detected by directly measuring the dies or the images of dies under toolmaker microscope. Measuring the dies of worn dentition directly under tool maker microscope enabled detection of wear progression more accurately than measuring die images obtained with Cercon Digital Machine. Conventional method was the least sensitive for tooth wear quantification and was unable to identify wear progression in most cases. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Fluorescence microscopy.

    PubMed

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

    2014-10-01

    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.

  6. Fluorescence Microscopy

    PubMed Central

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

    2016-01-01

    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. PMID:25275114

  7. Method and apparatus for chemical and topographical microanalysis

    NASA Technical Reports Server (NTRS)

    Kossakovski, Dmitri A. (Inventor); Baldeschwieler, John D. (Inventor); Beauchamp, Jesse L. (Inventor)

    2002-01-01

    A scanning probe microscope is combined with a laser induced breakdown spectrometer to provide spatially resolved chemical analysis of the surface correlated with the surface topography. Topographical analysis is achieved by scanning a sharp probe across the sample at constant distance from the surface. Chemical analysis is achieved by the means of laser induced breakdown spectroscopy by delivering pulsed laser radiation to the sample surface through the same sharp probe, and consequent collection and analysis of emission spectra from plasma generated on the sample by the laser radiation. The method comprises performing microtopographical analysis of the sample with a scanning probe, selecting a scanned topological site on the sample, generating a plasma plume at the selected scanned topological site, and measuring a spectrum of optical emission from the plasma at the selected scanned topological site. The apparatus comprises a scanning probe, a pulsed laser optically coupled to the probe, an optical spectrometer, and a controller coupled to the scanner, laser and spectrometer for controlling the operation of the scanner, laser and spectrometer. The probe and scanner are used for topographical profiling the sample. The probe is also used for laser radiation delivery to the sample for generating a plasma plume from the sample. Optical emission from the plasma plume is collected and delivered to the optical spectrometer so that analysis of emission spectrum by the optical spectrometer allows for identification of chemical composition of the sample at user selected sites.

  8. A high performance, cost-effective, open-source microscope for scanning two-photon microscopy that is modular and readily adaptable.

    PubMed

    Rosenegger, David G; Tran, Cam Ha T; LeDue, Jeffery; Zhou, Ning; Gordon, Grant R

    2014-01-01

    Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems.

  9. Wide-band acousto-optic deflectors for large field of view two-photon microscope.

    PubMed

    Jiang, Runhua; Zhou, Zhenqiao; Lv, Xiaohua; Zeng, Shaoqun

    2012-04-01

    Acousto-optic deflector (AOD) is an attractive scanner for two-photon microscopy because it can provide fast and versatile laser scanning and does not involve any mechanical movements. However, due to the small scan range of available AOD, the field of view (FOV) of the AOD-based microscope is typically smaller than that of the conventional galvanometer-based microscope. Here, we developed a novel wide-band AOD to enlarge the scan angle. Considering the maximum acceptable acoustic attenuation in the acousto-optic crystal, relatively lower operating frequencies and moderate aperture were adopted. The custom AOD was able to provide 60 MHz 3-dB bandwidth and 80% peak diffraction efficiency at 840 nm wavelength. Based on a pair of such AOD, a large FOV two-photon microscope was built with a FOV up to 418.5 μm (40× objective). The spatiotemporal dispersion was compensated simultaneously with a single custom-made prism. By means of dynamic power modulation, the variation of laser intensity within the FOV was reduced below 5%. The lateral and axial resolution of the system were 0.58-2.12 μm and 2.17-3.07 μm, respectively. Pollen grain images acquired by this system were presented to demonstrate the imaging capability at different positions across the entire FOV. © 2012 American Institute of Physics

  10. A High Performance, Cost-Effective, Open-Source Microscope for Scanning Two-Photon Microscopy that Is Modular and Readily Adaptable

    PubMed Central

    Rosenegger, David G.; Tran, Cam Ha T.; LeDue, Jeffery; Zhou, Ning; Gordon, Grant R.

    2014-01-01

    Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems. PMID:25333934

  11. Microwave Frequency Comb from a Semiconductor in a Scanning Tunneling Microscope.

    PubMed

    Hagmann, Mark J; Yarotski, Dmitry A; Mousa, Marwan S

    2017-04-01

    Quasi-periodic excitation of the tunneling junction in a scanning tunneling microscope, by a mode-locked ultrafast laser, superimposes a regular sequence of 15 fs pulses on the DC tunneling current. In the frequency domain, this is a frequency comb with harmonics at integer multiples of the laser pulse repetition frequency. With a gold sample the 200th harmonic at 14.85 GHz has a signal-to-noise ratio of 25 dB, and the power at each harmonic varies inversely with the square of the frequency. Now we report the first measurements with a semiconductor where the laser photon energy must be less than the bandgap energy of the semiconductor; the microwave frequency comb must be measured within 200 μm of the tunneling junction; and the microwave power is 25 dB below that with a metal sample and falls off more rapidly at the higher harmonics. Our results suggest that the measured attenuation of the microwave harmonics is sensitive to the semiconductor spreading resistance within 1 nm of the tunneling junction. This approach may enable sub-nanometer carrier profiling of semiconductors without requiring the diamond nanoprobes in scanning spreading resistance microscopy.

  12. Direct laser sintered WC-10Co/Cu nanocomposites

    NASA Astrophysics Data System (ADS)

    Gu, Dongdong; Shen, Yifu

    2008-04-01

    In the present work, the direct metal laser sintering (DMLS) process was used to prepare the WC-Co/Cu nanocomposites in bulk form. The WC reinforcing nanoparticles were added in the form of WC-10 wt.% Co composite powder. The microstructural features and mechanical properties of the laser-sintered sample were characterized by X-ray diffraction (XRD), atomic force microscope (AFM), scanning electron microscope (SEM), energy dispersive X-ray spectroscope (EDX), and nanoindentation tester. It showed that the original nanometric nature of the WC reinforcing particulates was well retained without appreciable grain growth after laser processing. A homogeneous distribution of the WC reinforcing nanoparticles with a coherent particulate/matrix interfacial bonding was obtained in the laser-sintered structure. The 94.3% dense nanocomposites have a dynamic nanohardness of 3.47 GPa and a reduced elastic modulus of 613.42 GPa.

  13. Surface Modifications with Laser Synthesized Mo Modified Coating

    NASA Astrophysics Data System (ADS)

    Sun, Lu; Chen, Hao; Liu, Bo

    2013-01-01

    Mg-Cu-Al was first used to improve the surface performance of TA15 titanium alloys by means of laser cladding technique. The synthesis of hard composite coating on TA15 titanium alloy by laser cladding of Mg-Cu-Al-B4C/Mo pre-placed powders was investigated by means of scanning electron microscope, energy dispersive spectrometer and high resolution transmission electron microscope. Experimental results indicated that such composite coating mainly consisted of TiB2, TiB, TiC, Ti3Al and AlCuMg. Compared with TA15 alloy substrate, an improvement of wear resistance was observed for this composite coating due to the actions of fine grain, amorphous and hard phase strengthening.

  14. Corrosion performance of 7075 alloy under laser heat treatment

    NASA Astrophysics Data System (ADS)

    Liu, Tong; Su, Ruiming; Qu, Yingdong; Li, Rongde

    2018-05-01

    Microstructure, exfoliation corrosion (EXCO), intergranular corrosion (IGC) and potentidynamic polarization test of the 7075 aluminum alloy after retrogression and re-aging (RRA) treatment, and laser retrogression and re-aging (LRRA), respectively, were studied by using scanning electron microscope, and transmission electron microscope (TEM). The results show that after pre-aging, laser treatment (650 W, 2 mm s‑1) and re-aging a lot of matrix precipitates of alloy were precipitated again. The semi-continuous grain boundary precipitates and the wider precipitate-free zones (PFZ) improve the corrosion resistance of the alloy. The corrosion properties of the alloy after LRRA (650 W, 2 mm s‑1) treatment are better than that after RRA treatment.

  15. MEMS-based handheld confocal microscope for in-vivo skin imaging

    PubMed Central

    Arrasmith, Christopher L.; Dickensheets, David L.; Mahadevan-Jansen, Anita

    2010-01-01

    This paper describes a handheld laser scanning confocal microscope for skin microscopy. Beam scanning is accomplished with an electromagnetic MEMS bi-axial micromirror developed for pico projector applications, providing 800x600 (SVGA) resolution at 56 frames per second. The design uses commercial objective lenses with an optional hemisphere front lens, operating with a range of numerical aperture from NA=0.35 to NA=1.1 and corresponding diagonal field of view ranging from 653 μm to 216 μm. Using NA=1.1 and a laser wavelength of 830 nm we measured the axial response to be 1.14 μm full width at half maximum, with a corresponding 10%-90% lateral edge response of 0.39 μm. Image examples showing both epidermal and dermal features including capillary blood flow are provided. These images represent the highest resolution and frame rate yet achieved for tissue imaging with a MEMS bi-axial scan mirror. PMID:20389391

  16. Sealing ability of three root-end filling materials prepared using an erbium: Yttrium aluminium garnet laser and endosonic tip evaluated by confocal laser scanning microscopy

    PubMed Central

    Nanjappa, A Salin; Ponnappa, KC; Nanjamma, KK; Ponappa, MC; Girish, Sabari; Nitin, Anita

    2015-01-01

    Aims: (1) To compare the sealing ability of mineral trioxide aggregate (MTA), Biodentine, and Chitra-calcium phosphate cement (CPC) when used as root-end filling, evaluated under confocal laser scanning microscope using Rhodamine B dye. (2) To evaluate effect of ultrasonic retroprep tip and an erbium:yttrium aluminium garnet (Er:YAG) laser on the integrity of three different root-end filling materials. Materials and Methods: The root canals of 80 extracted teeth were instrumented and obturated with gutta-percha. The apical 3 mm of each tooth was resected and 3 mm root-end preparation was made using ultrasonic tip (n = 30) and Er:YAG laser (n = 30). MTA, Biodentine, and Chitra-CPC were used to restore 10 teeth each. The samples were coated with varnish and after drying, they were immersed in Rhodamine B dye for 24 h. The teeth were then rinsed, sectioned longitudinally, and observed under confocal laser scanning microscope. Statistical Analysis Used: Data were analyzed using one-way analysis of variance (ANOVA) and a post-hoc Tukey's test at P < 0.05 (R software version 3.1.0). Results: Comparison of microleakage showed maximum peak value of 0.45 mm for Biodentine, 0.85 mm for MTA, and 1.05 mm for Chitra-CPC. The amount of dye penetration was found to be lesser in root ends prepared using Er:YAG laser when compared with ultrasonics, the difference was found to be statistically significant (P < 0.05). Conclusions: Root-end cavities prepared with Er:YAG laser and restored with Biodentine showed superior sealing ability compared to those prepared with ultrasonics. PMID:26180420

  17. First results for custom-built low-temperature (4.2 K) scanning tunneling microscope/molecular beam epitaxy and pulsed laser epitaxy system designed for spin-polarized measurements

    NASA Astrophysics Data System (ADS)

    Foley, Andrew; Alam, Khan; Lin, Wenzhi; Wang, Kangkang; Chinchore, Abhijit; Corbett, Joseph; Savage, Alan; Chen, Tianjiao; Shi, Meng; Pak, Jeongihm; Smith, Arthur

    2014-03-01

    A custom low-temperature (4.2 K) scanning tunneling microscope system has been developed which is combined directly with a custom molecular beam epitaxy facility (and also including pulsed laser epitaxy) for the purpose of studying surface nanomagnetism of complex spintronic materials down to the atomic scale. For purposes of carrying out spin-polarized STM measurements, the microscope is built into a split-coil, 4.5 Tesla superconducting magnet system where the magnetic field can be applied normal to the sample surface; since, as a result, the microscope does not include eddy current damping, vibration isolation is achieved using a unique combination of two stages of pneumatic isolators along with an acoustical noise shield, in addition to the use of a highly stable as well as modular `Pan'-style STM design with a high Q factor. First 4.2 K results reveal, with clear atomic resolution, various reconstructions on wurtzite GaN c-plane surfaces grown by MBE, including the c(6x12) on N-polar GaN(0001). Details of the system design and functionality will be presented.

  18. Phase Sensitive Demodulation in Multiphoton Microscopy

    NASA Astrophysics Data System (ADS)

    Fisher, Walt G.; Piston, David W.; Wachter, Eric A.

    2002-06-01

    Multiphoton laser scanning microscopy offers advantages in depth of penetration into intact samples over other optical sectioning techniques. To achieve these advantages it is necessary to detect the emitted light without spatial filtering. In this nondescanned (nonconfocal) approach, ambient room light can easily contaminate the signal, forcing experiments to be performed in absolute darkness. For multiphoton microscope systems employing mode-locked lasers, signal processing can be used to reduce such problems by taking advantage of the pulsed characteristics of such lasers. Specifically, by recovering fluorescence generated at the mode-locked frequency, interference from stray light and other ambient noise sources can be significantly reduced. This technology can be adapted to existing microscopes by inserting demodulation circuitry between the detector and data collection system. The improvement in signal-to-noise ratio afforded by this approach yields a more robust microscope system and opens the possibility of moving multiphoton microscopy from the research lab to more demanding settings, such as the clinic.

  19. Changes in the emission properties of metallic targets upon exposure to repetitively pulsed laser radiation

    NASA Astrophysics Data System (ADS)

    Konov, V. I.; Pimenov, S. M.; Prokhorov, A. M.; Chapliev, N. I.

    1988-02-01

    A scanning electron microscope and a repetitively pulsed CO2 laser are used to reveal the relationships which govern the correlation of the transforming metal surface microrelief with the emission of charged particles and the surface luminescence upon exposure to multipulse laser focusing. It is shown that the effect of sorption and laser-stimulated desorption on the emission signals can manifest itself in different ways depending on the current oscillation mode in the target-vacuum chamber circuit.

  20. Special Features of the Optical Absorption and Photoconductivity of Indium Monoselenide Upon Laser Excitation

    NASA Astrophysics Data System (ADS)

    Kyazym-Zade, A. G.; Salmanov, V. M.; Guseinov, A. G.; Mamedov, R. M.; Salmanova, A. A.; Akhmedova, F. Sh.

    2018-02-01

    The successive ionic layer adsorption and reaction (SILAR) method is used to prepare InSe thin films and InSe nanoparticles. Shapes and sizes of the obtained nanoparticles are investigated using a scanning electron microscope and an atomic force microscope. The main parameters of the examined structures, nanoparticle sizes (4-20 nm), and band gap ( E g = 1.60 eV) for nanoparticles with the least sizes are determined. Superfast (1.5·10-8 s) photocurrent relaxation and stimulated emission with line half-width of 8 Å have been observed upon exposure to laser radiation.

  1. An in-vitro evaluation of the effect of 980 nm diode laser irradiation on intra-canal dentin surface and dentinal tubule openings after biomechanical preparation: Scanning electron microscopic study

    PubMed Central

    Jhingan, Pulkit; Sandhu, Meera; Jindal, Garima; Goel, Deepti; Sachdev, Vinod

    2015-01-01

    Context: Very recently, diode laser has been used for disinfecting the root canals in endodontic treatment and increasing its success rate and longevity utilizing the thermal effect of laser on surrounding tissues. Aims: The aim of this study is to evaluate the effect of 980 nm laser irradiation on intra-canal dentin surface – scanning electron microscopic (SEM) - in-vitro study. Methods: A total of 40 single-rooted freshly extracted permanent teeth were collected. Teeth were sectioned at the cemento-enamel junction using diamond disc. Root canals of all samples were prepared using hand ProTaper, which were randomly assigned into two groups (n = 20 each). Group 1: Receiving no treatment after biomechanical preparation; Group 2: 980 nm diode laser-treated root canals. Teeth were prepared for SEM analysis to check the size of intra-canal dentinal tubule openings. Statistical Analysis Used: Data were analyzed using SPSS V.16 software and compared using Levene's and independent t-test. Results: On statistical analysis, width of intracanal dentinal tubule openings in Group 1 (control) was significantly higher than those observed in Group 2 (diode laser-treated) (P < 0.001). Conclusion: This study showed that the application of 980 nm diode laser on intra-radicular dentin resulted in ultrastructural alterations resulting in melting of dentin. PMID:26097338

  2. Scanning electron microscope comparative surface evaluation of glazed-lithium disilicate ceramics under different irradiation settings of Nd:YAG and Er:YAG lasers.

    PubMed

    Viskic, Josko; Jokic, Drazen; Jakovljevic, Suzana; Bergman, Lana; Ortolan, Sladana Milardovic; Mestrovic, Senka; Mehulic, Ketij

    2018-01-01

    To evaluate the surface of glazed lithium disilicate dental ceramics after irradiation under different irradiation settings of Nd:YAG and Er:YAG lasers using a scanning electron microscope (SEM). Three glazed-press lithium disilicate ceramic discs were treated with HF, Er:YAG, and Nd:YAG, respectively. The laser-setting variables tested were laser mode, repetition rate (Hz), power (W), time of exposure (seconds), and laser energy (mJ). Sixteen different variable settings were tested for each laser type, and all the samples were analyzed by SEM at 500× and 1000× magnification. Surface analysis of the HF-treated sample showed a typical surface texture with a homogenously rough pattern and exposed ceramic crystals. Er:YAG showed no effect on the surface under any irradiation setting. The surface of Nd:YAG-irradiated samples showed cracking, melting, and resolidifying of the ceramic glaze. These changes became more pronounced as the power increased. At the highest power setting (2.25 W), craters on the surface with large areas of melted or resolidified glaze surrounded by globules were visible. However, there was little to no exposure of ceramic crystals or visible regular surface roughening. Neither Er:YAG nor Nd:YAG dental lasers exhibited adequate surface modification for bonding of orthodontic brackets on glazed lithium disilicate ceramics compared with the control treated with 9.5% HF.

  3. Control and acquisition systems for new scanning transmission x-ray microscopes at Advanced Light Source (abstract)

    NASA Astrophysics Data System (ADS)

    Tyliszczak, T.; Hitchcock, P.; Kilcoyne, A. L. D.; Ade, H.; Hitchcock, A. P.; Fakra, S.; Steele, W. F.; Warwick, T.

    2002-03-01

    Two new scanning x-ray transmission microscopes are being built at beamline 5.3.2 and beamline 7.0 of the Advanced Light Source that have novel aspects in their control and acquisition systems. Both microscopes use multiaxis laser interferometry to improve the precision of pixel location during imaging and energy scans as well as to remove image distortions. Beam line 5.3.2 is a new beam line where the new microscope will be dedicated to studies of polymers in the 250-600 eV energy range. Since this is a bending magnet beam line with lower x-ray brightness than undulator beam lines, special attention is given to the design not only to minimize distortions and vibrations but also to optimize the controls and acquisition to improve data collection efficiency. 5.3.2 microscope control and acquisition is based on a PC computer running WINDOWS 2000. All mechanical stages are moved by stepper motors with rack mounted controllers. A dedicated counter board is used for counting and timing and a multi-input/output board is used for analog acquisition and control of the focusing mirror. A three axis differential laser interferometer is being used to improve stability and precision by careful tracking of the relative positions of the sample and zone plate. Each axis measures the relative distance between a mirror placed on the sample stage and a mirror attached to the zone plate holder. Agilent Technologies HP 10889A servo-axis interferometer boards are used. While they were designed to control servo motors, our tests show that they can be used to directly control the piezo stage. The use of the interferometer servo-axis boards provides excellent point stability for spectral measurements. The interferometric feedback also provides active vibration isolation which reduces deleterious impact of mechanical vibrations up to 20-30 Hz. It also can improve the speed and precision of image scans. Custom C++ software has been written to provide user friendly control of the microscope and integration with visual light microscopy indexing of the samples. The beam line 7.0 microscope upgrade is a new design which will replace the existing microscope. The design is similar to that of beam line 5.3.2, including interferometric position encoding. However the acquisition and control is based on VXI systems, a Sun computer, and LABVIEW™ software. The main objective of the BL 7.0 microscope upgrade is to achieve precise image scans at very high speed (pixel dwells as short as 10 μs) to take full advantage of the high brightness of the 7.0 undulator beamline. Results of tests and a discussion of the benefits of our scanning microscope designs will be presented.

  4. Laser Cladding of Ti-6Al-4V Alloy with Ti-Al2O3 Coating for Biomedical Applications

    NASA Astrophysics Data System (ADS)

    Mthisi, A.; Popoola, A. P. I.; Adebiyi, D. I.; Popoola, O. M.

    2018-05-01

    The indispensable properties of Ti-6Al-4V alloy coupled with poor tribological properties and delayed bioactivity make it a subject of interest to explore in biomedical application. A quite number of numerous coatings have been employed on titanium alloys, with aim to overcome the poor properties exhibited by this alloy. In this work, the possibility of laser cladding different ad-mixed powders (Ti - 5 wt.% Al2O3 and Ti - 8wt.% Al2O3) on Ti-6Al-4V at various laser scan speed (0.6 and 0.8 m/min) were investigated. The microstructure, phase constituents and corrosion of the resultant coatings were characterized by scanning electron microscope (SEM), Optical microscope, X-Ray diffractometer (XRD) and potentiostat respectively. The electrochemical behaviour of the produced coatings was studied in a simulated body fluid (Hanks solution). The microstructural results show that a defect free coating is achieved at low scan speed and ad-mixed of Ti-5 wt. % Al2O3. Cladding of Ti - Al2O3 improved the corrosion resistance of Ti-6Al-4V alloy regardless of varying neither scan speed nor ad-mixed percentage. However, Ti-5 wt.% Al2O3 coating produced at low scan speed revealed the highest corrosion resistance among the coatings due to better quality coating layer. Henceforth, this coating may be suitable for biomedical applications.

  5. Miniaturized fiber-coupled confocal fluorescence microscope with an electrowetting variable focus lens using no moving parts

    PubMed Central

    Ozbay, Baris N.; Losacco, Justin T.; Cormack, Robert; Weir, Richard; Bright, Victor M.; Gopinath, Juliet T.; Restrepo, Diego; Gibson, Emily A.

    2015-01-01

    We report a miniature, lightweight fiber-coupled confocal fluorescence microscope that incorporates an electrowetting variable focus lens to provide axial scanning for full three-dimensional (3D) imaging. Lateral scanning is accomplished by coupling our device to a laser-scanning confocal microscope through a coherent imaging fiber-bundle. The optical components of the device are combined in a custom 3D-printed adapter with an assembled weight of <2 g that can be mounted onto the head of a mouse. Confocal sectioning provides an axial resolution of ~12 µm and an axial scan range of ~80 µm. The lateral field-of-view is 300 µm, and the lateral resolution is 1.8 µm. We determined these parameters by imaging fixed sections of mouse neuronal tissue labeled with green fluorescent protein (GFP) and fluorescent bead samples in agarose gel. To demonstrate viability for imaging intact tissue, we resolved multiple optical sections of ex vivo mouse olfactory nerve fibers expressing yellow fluorescent protein (YFP). PMID:26030555

  6. Two-Photon Fluorescence Correlation Spectroscopy

    NASA Technical Reports Server (NTRS)

    Zimmerli, Gregory A.; Fischer, David G.

    2002-01-01

    We will describe a two-photon microscope currently under development at the NASA Glenn Research Center. It is composed of a Coherent Mira 900 tunable, pulsed Titanium:Sapphire laser system, an Olympus Fluoview 300 confocal scanning head, and a Leica DM IRE inverted microscope. It will be used in conjunction with a technique known as fluorescence correlation spectroscopy (FCS) to study intracellular protein dynamics. We will briefly explain the advantages of the two-photon system over a conventional confocal microscope, and provide some preliminary experimental results.

  7. Ultrahigh resolution multicolor colocalization of single fluorescent probes

    DOEpatents

    Weiss, Shimon; Michalet, Xavier; Lacoste, Thilo D.

    2005-01-18

    A novel optical ruler based on ultrahigh-resolution colocalization of single fluorescent probes is described. Two unique families of fluorophores are used, namely energy-transfer fluorescent beads and semiconductor nanocrystal (NC) quantum dots, that can be excited by a single laser wavelength but emit at different wavelengths. A novel multicolor sample-scanning confocal microscope was constructed which allows one to image each fluorescent light emitter, free of chromatic aberrations, by scanning the sample with nanometer scale steps using a piezo-scanner. The resulting spots are accurately localized by fitting them to the known shape of the excitation point-spread-function of the microscope.

  8. Use of scanning near-field optical microscope with an aperture probe for detection of luminescent nanodiamonds

    NASA Astrophysics Data System (ADS)

    Shershulin, V. A.; Samoylenko, S. R.; Shenderova, O. A.; Konov, V. I.; Vlasov, I. I.

    2017-02-01

    The suitability of scanning near-field optical microscopy (SNOM) to image photoluminescent diamond nanoparticles with nanoscale resolution is demonstrated. Isolated diamond nanocrystals with an average size of 100 nm, containing negatively charged nitrogen-vacancy (NV-) centers, were chosen as tested material. The NV- luminescence was stimulated by continuous 532 nm laser light. Sizes of analyzed crystallites were monitored by an atomic force microscope. The lateral resolution of the order of 100 nm was reached in SNOM imaging of diamond nanoparticles using 150 nm square aperture of the probe.

  9. Morphology of Er:YAG-laser-treated root surfaces

    NASA Astrophysics Data System (ADS)

    Keller, Ulrich; Stock, Karl; Hibst, Raimund

    1997-12-01

    From previous studies it could be demonstrated that an efficient ablation of dental calculus is possible using an Er:YAG laser with a special contact fiber tip. After improving of the design and the efficiency of light transmission of the contact tip laser treated tooth root surfaces were investigated due to morphological changes in comparison to conventional root scaling and planing. Surface modifications were observed histologically under the light microscope and by means of a Scanning Electron Microscope. During laser treatment the intrapulpal temperature increase was measured. The results show that the improved contact tip a microstructured surface can be generated, which shows no signs of thermal effects even when a laser pulse repetition rate of 15 Hz was used. Temperature increase was limited to 4 K at a repetition rate of 10 Hz and to 5.5 K at a repetition rate of 15 Hz.

  10. Research on Microstructure and Properties of Welded Joint of High Strength Steel

    NASA Astrophysics Data System (ADS)

    Zhu, Pengxiao; Li, Yi; Chen, Bo; Ma, Xuejiao; Zhang, Dongya; Tang, Cai

    2018-01-01

    BS960 steel plates were welded by Laser-MAG and MAG. The microstructure and properties of the welded joints were investigated by optical microscope, micro-hardness tester, universal tensile testing machine, impact tester, scanning electron microscope (SEM) and fatigue tester. By a series of experiments, the following results were obtained: The grain size of the coarse grain zone with Laser-MAG welded joint is 20μm, and that with MAG welded joint is about 32μm, both of the fine grain region are composed of fine lath martensite and granular bainite; the width of the heat affected region with Laser-MAG is lower than that with MAG. The strength and impact energy of welded joints with Laser-MAG is higher than that with MAG. The conditioned fatigue limit of welded joint with Laser-MAG is 280MPa; however, the conditioned fatigue limit of welded joint with MAG is 250MPa.

  11. Scalable patterning using laser-induced shock waves

    NASA Astrophysics Data System (ADS)

    Ilhom, Saidjafarzoda; Kholikov, Khomidkhodza; Li, Peizhen; Ottman, Claire; Sanford, Dylan; Thomas, Zachary; San, Omer; Karaca, Haluk E.; Er, Ali O.

    2018-04-01

    An advanced direct imprinting method with low cost, quick, and minimal environmental impact to create a thermally controllable surface pattern using the laser pulses is reported. Patterned microindents were generated on Ni50Ti50 shape memory alloys and aluminum using an Nd: YAG laser operating at 1064 nm combined with a suitable transparent overlay, a sacrificial layer of graphite, and copper grid. Laser pulses at different energy densities, which generate pressure pulses up to a few GPa on the surface, were focused through the confinement medium, ablating the copper grid to create plasma and transferring the grid pattern onto the surface. Scanning electron microscope and optical microscope images show that various patterns were obtained on the surface with high fidelity. One-dimensional profile analysis indicates that the depth of the patterned sample initially increases with the laser energy and later levels off. Our simulations of laser irradiation process also confirm that high temperature and high pressure could be generated when the laser energy density of 2 J/cm2 is used.

  12. Performance of a three-dimensional-printed microscanner in a laser scanning microscopy application

    NASA Astrophysics Data System (ADS)

    Oyman, Hilmi Artun; Gokdel, Yigit Daghan; Ferhanoglu, Onur; Yalcinkaya, Arda Deniz

    2018-04-01

    A magnetically actuated microscanner is used in a laser scanning microscopy application. Stress distribution along the circular-profiled flexure is compared with a rectangular counterpart in finite-element environment. Magnetic actuation mechanism of the scanning unit is explained in detail. Moreover, reliability of the scanner is tested for 3×106 cycle. The scanning device is designed to meet a confocal microscopy application providing 100 μm×100 μm field of view and <3-μm lateral resolution. The resonance frequencies of the device were analytically modeled, where we obtained 130- and 268-Hz resonance values for the out-of-plane and torsion modes, respectively. The scanning device provided an optical scan angle about 2.5 deg for 170-mA drive current, enabling the desired field of view for our custom built confocal microscope setup. Finally, imaging experiments were conducted on a resolution target, showcasing the desired scan area and resolution.

  13. Thermal radiation scanning tunnelling microscopy

    NASA Astrophysics Data System (ADS)

    de Wilde, Yannick; Formanek, Florian; Carminati, Rémi; Gralak, Boris; Lemoine, Paul-Arthur; Joulain, Karl; Mulet, Jean-Philippe; Chen, Yong; Greffet, Jean-Jacques

    2006-12-01

    In standard near-field scanning optical microscopy (NSOM), a subwavelength probe acts as an optical `stethoscope' to map the near field produced at the sample surface by external illumination. This technique has been applied using visible, infrared, terahertz and gigahertz radiation to illuminate the sample, providing a resolution well beyond the diffraction limit. NSOM is well suited to study surface waves such as surface plasmons or surface-phonon polaritons. Using an aperture NSOM with visible laser illumination, a near-field interference pattern around a corral structure has been observed, whose features were similar to the scanning tunnelling microscope image of the electronic waves in a quantum corral. Here we describe an infrared NSOM that operates without any external illumination: it is a near-field analogue of a night-vision camera, making use of the thermal infrared evanescent fields emitted by the surface, and behaves as an optical scanning tunnelling microscope. We therefore term this instrument a `thermal radiation scanning tunnelling microscope' (TRSTM). We show the first TRSTM images of thermally excited surface plasmons, and demonstrate spatial coherence effects in near-field thermal emission.

  14. Adapting a compact confocal microscope system to a two-photon excitation fluorescence imaging architecture.

    PubMed

    Diaspro, A; Corosu, M; Ramoino, P; Robello, M

    1999-11-01

    Within the framework of a national National Institute of Physics of Matter (INFM) project, we have realised a two-photon excitation (TPE) fluorescence microscope based on a new generation commercial confocal scanning head. The core of the architecture is a mode-locked Ti:Sapphire laser (Tsunami 3960, Spectra Physics Inc., Mountain View, CA) pumped by a high-power (5 W, 532 nm) laser (Millennia V, Spectra Physics Inc.) and an ultracompact confocal scanning head, Nikon PCM2000 (Nikon Instruments, Florence, Italy) using a single-pinhole design. Three-dimensional point-spread function has been measured to define spatial resolution performances. The TPE microscope has been used with a wide range of excitable fluorescent molecules (DAPI, Fura-2, Indo-1, DiOC(6)(3), fluoresceine, Texas red) covering a single photon spectral range from UV to green. An example is reported on 3D imaging of the helical structure of the sperm head of the Octopus Eledone cirrhosa labelled with an UV excitable dye, i.e., DAPI. The system can be easily switched for operating both in conventional and two-photon mode. Copyright 1999 Wiley-Liss, Inc.

  15. Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

    PubMed Central

    Lim, Daryl; Ford, Tim N.; Chu, Kengyeh K.; Mertz, Jerome

    2011-01-01

    We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish. PMID:21280920

  16. Optically sectioned in vivo imaging with speckle illumination HiLo microscopy.

    PubMed

    Lim, Daryl; Ford, Tim N; Chu, Kengyeh K; Mertz, Jerome

    2011-01-01

    We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

  17. Optically sectioned in vivo imaging with speckle illumination HiLo microscopy

    NASA Astrophysics Data System (ADS)

    Lim, Daryl; Ford, Tim N.; Chu, Kengyeh K.; Mertz, Jerome

    2011-01-01

    We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

  18. Mirrorlike pulsed laser deposited tungsten thin film.

    PubMed

    Mostako, A T T; Rao, C V S; Khare, Alika

    2011-01-01

    Mirrorlike tungsten thin films on stainless steel substrate deposited via pulsed laser deposition technique in vacuum (10(-5) Torr) is reported, which may find direct application as first mirror in fusion devices. The crystal structure of tungsten film is analyzed using x-ray diffraction pattern, surface morphology of the tungsten films is studied with scanning electron microscope and atomic force microscope. The film composition is identified using energy dispersive x-ray. The specular and diffuse reflectivities with respect to stainless steel substrate of the tungsten films are recorded with FTIR spectra. The thickness and the optical quality of pulsed laser deposition deposited films are tested via interferometric technique. The reflectivity is approaching about that of the bulk for the tungsten film of thickness ∼782 nm.

  19. Effects of Laser Shock Processing on Morphologies and Mechanical Properties of ANSI 304 Stainless Steel Weldments Subjected to Cavitation Erosion

    PubMed Central

    Zhang, Lei; Lu, Jin-Zhong; Zhang, Yong-Kang; Ma, Hai-Le; Luo, Kai-Yu; Dai, Feng-Ze

    2017-01-01

    Effects of laser shock processing (LSP) on the cavitation erosion resistance of laser weldments were investigated by optical microscope (OM), scanning electron microscope (SEM) observations, roughness tester, micro hardness tester, and X-ray diffraction (XRD) technology. The morphological microstructures were characterized. Cumulative mass loss, incubation period, erosion rate, and damaged surface areas were monitored during cavitation erosion. Surface roughness, micro-hardness, and residual stress were measured in different zones. Results showed that LSP could improve the damage of morphological microstructures and mechanical properties after cavitation erosion. The compressive residual stresses were generated during the process of LSP, which was an effective guarantee for the improvement of the above mentioned properties. PMID:28772652

  20. A portable confocal hyperspectral microscope without any scan or tube lens and its application in fluorescence and Raman spectral imaging

    NASA Astrophysics Data System (ADS)

    Li, Jingwei; Cai, Fuhong; Dong, Yongjiang; Zhu, Zhenfeng; Sun, Xianhe; Zhang, Hequn; He, Sailing

    2017-06-01

    In this study, a portable confocal hyperspectral microscope is developed. In traditional confocal laser scanning microscopes, scan lens and tube lens are utilized to achieve a conjugate relationship between the galvanometer and the back focal plane of the objective, in order to achieve a better resolution. However, these lenses make it difficult to scale down the volume of the system. In our portable confocal hyperspectral microscope (PCHM), the objective is placed directly next to the galvomirror. Thus, scan lens and tube lens are not included in our system and the size of this system is greatly reduced. Furthermore, the resolution is also acceptable in many biomedical and food-safety applications. Through reducing the optical length of the system, the signal detection efficiency is enhanced. This is conducive to realizing both the fluorescence and Raman hyperspectral imaging. With a multimode fiber as a pinhole, an improved image contrast is also achieved. Fluorescent spectral images for HeLa cells/fingers and Raman spectral images of kumquat pericarp are present. The spectral resolution and spatial resolutions are about 0.4 nm and 2.19 μm, respectively. These results demonstrate that this portable hyperspectral microscope can be used in in-vivo fluorescence imaging and in situ Raman spectral imaging.

  1. Effect of diode laser and ultrasonics with and without ethylenediaminetetraacetic acid on smear layer removal from the root canals: A scanning electron microscope study.

    PubMed

    Amin, Khalid; Masoodi, Ajaz; Nabi, Shahnaz; Ahmad, Parvaiz; Farooq, Riyaz; Purra, Aamir Rashid; Ahangar, Fayaz Ahmad

    2016-01-01

    To evaluate the effect of diode laser and ultrasonics with and without ethylenediaminetetraacetic acid (EDTA) on the smear layer removal from root canals. A total of 120 mandibular premolars were decoronated to working the length of 12 mm and prepared with protaper rotary files up to size F3. Group A canals irrigated with 1 ml of 3% sodium hypochlorite (NaOCl) followed by 3 ml of 3% NaOCl. Group B canals irrigated with 1 ml of 17% EDTA followed by 3 ml of 3% NaOCl. Group C canals lased with a diode laser. Group D canals were initially irrigated with 0.8 ml of 17% EDTA the remaining 0.2 ml was used to fill the root canals, and diode laser application was done. Group E canals were irrigated with 1 ml distilled water with passive ultrasonic activation, followed by 3 ml of 3% NaOCl. Group F canals were irrigated with 1 ml EDTA with passive ultrasonic activation, followed by 3 ml of 3% NaOCl. Scanning electron microscope examination of canals was done for remaining smear layer at coronal middle and apical third levels. Ultrasonics with EDTA had the least smear layer scores. Diode laser alone performed significantly better than ultrasonics.

  2. In-situ Crystallization of Highly Volatile Commercial Mold Flux Using an Isolated Observation System in the Confocal Laser Scanning Microscope

    NASA Astrophysics Data System (ADS)

    Park, Jun-Yong; Ryu, Jae Wook; Sohn, Il

    2014-08-01

    The in situ crystallization behavior of highly volatile commercial mold fluxes for medium carbon steels was investigated using the confocal laser scanning microscope (CLSM) equipped with an optimized isolated observation system. The highly volatile compounds of the mold flux were suppressed during heating allowing direct observation in the CLSM. Cooling rates of 25, 50, 100, 400, and 800 K/min were incorporated and continuous cooling transformation (CCT) diagrams of 4 different commercial mold fluxes for medium carbon steels were developed. Identification of the crystalline phase was conducted with XRD and SEM-EDS analysis. A cuspidine crystalline was observed in all samples at various cooling rates. With higher basicity, CaF2, and NaF, the crystallization of the fluxes was enhanced according to the CCT diagram. As the slag structure becomes depolymerized, the diffusion rate of the cathodic ions seems to increase.

  3. INTERACTION OF LASER RADIATION WITH MATTER AND OTHER LASER APPLICATIONS: Changes in the emission properties of metal targets during pulse-periodic laser irradiation

    NASA Astrophysics Data System (ADS)

    Konov, Vitalii I.; Pimenov, S. M.; Prokhorov, A. M.; Chapliev, N. I.

    1988-02-01

    A scanning electron microscope was used with a pulse-periodic CO2 laser to discover the laws governing the correlation of the modified microrelief of metal surfaces, subjected to the action of multiple laser pulses, with the emission of charged particles and the luminescence of the irradiated zone. It was established that the influence of sorption and laser-induced desorption on the emission signals may be manifested differently depending on the regime of current generation in the "target-vacuum chamber" circuit.

  4. Comparative scanning electron microscope analysis of diode laser and desensitizing toothpastes for evaluation of efficacy of dentinal tubular occlusion.

    PubMed

    Reddy, Guntakala Vikram; Akula, Sushma; Malgikar, Suryakanth; Babu, Palaparthy Raja; Reddy, Gooty Jagadish; Josephin, Johnson Juliet

    2017-01-01

    The present study aims to evaluate the efficacy of diode laser alone and in combination with desensitizing toothpastes in occluding dentinal tubules (both partially occluded and completely occluded tubules) by scanning electron microscope (SEM). Fifty human teeth were extracted, cervical cavities were prepared and etched with 17% ethylenediaminetetraacetic acid, and smear layer was removed to expose the tubules. The teeth were divided into five groups: Group I - Application of NovaMin-formulated toothpaste, Group II - Application of Pro-Argin ™ -formulated toothpaste, Group III - Application of diode laser in noncontact mode, Group IV - NovaMin-formulated toothpaste followed by laser irradiation, and Group V - Pro-Argin ™ -formulated toothpaste followed by laser irradiation. After treatment, quantitative analysis of occluded dentinal tubules was done by SEM analysis. The mean values of percentages of total occlusion of dentinal tubules in Groups I, II, III, IV, and V were 92.73% ± 1.38, 90.67% ± 1.86, 96.57% ± 0.64, 97.3% ± 0.68, and 96.9% ± 6.08, respectively. Addition of diode laser (Groups III, IV, and V) yielded a significant occlusion of the dentinal tubules when compared to desensitizing toothpastes alone (Groups I and II). Diode laser (Group III) has shown more efficacy in occluding dentinal tubules when compared with desensitizing toothpastes which was statistically significant ( P < 0.05). Among the five groups, NovaMin + diode laser (Group IV) showed the highest percentage of occluded dentinal tubules.

  5. High-power direct green laser oscillation of 598 mW in Pr(3+)-doped waterproof fluoroaluminate glass fiber excited by two-polarization-combined GaN laser diodes.

    PubMed

    Nakanishi, Jun; Horiuchi, Yuya; Yamada, Tsuyoshi; Ishii, Osamu; Yamazaki, Masaaki; Yoshida, Minoru; Fujimoto, Yasushi

    2011-05-15

    We demonstrated a high-power and highly efficient Pr-doped waterproof fluoride glass fiber laser at 522.2 nm excited by two-polarization-combined GaN laser diodes and achieved a subwatt output power of 598 mW and slope efficiency of 43.0%. This system will enable us to make a vivid laser display, a photocoagulation laser for eye surgery, a color confocal scanning laser microscope, and an effective laser for material processing. Direct visible ultrashort pulse generation is also expected. © 2011 Optical Society of America

  6. Epiphany sealer penetration into dentinal tubules: Confocal laser scanning microscopic study.

    PubMed

    Ravi, S V; Nageswar, Rao; Swapna, Honwad; Sreekant, Puthalath; Ranjith, Madhavan; Mahidhar, Surabhi

    2014-03-01

    The aim of the following study was to evaluate the percentage and average depth of epiphany sealer penetration into dentinal tubules among the coronal, middle and apical thirds of the root using the confocal laser scanning microscopy (CLSM). A total of 10 maxillary central incisors were prepared and obturated with Resilon-Epiphany system. Sealer was mixed with fluorescent rhodamine B isothiyocyanate dye for visibility under confocal microscope. Teeth were cross-sectioned into coronal, middle and apical sections-2 mm thick. Sections were observed under CLSM. Images were analyzed for percentage and average depth of sealer penetration into dentinal tubules using the lasso tool in Adobe Photoshop CS3 (Adobe systems incorporated, San jose, CA) and laser scanning microscopy (LSM 5) image analyzer. One-way analysis of variance with Student Neuman Keuls post hoc tests, Kruskal-Wallis test and Wilcoxon signed-rank post hoc tests. The results showed that a higher percentage of sealer penetration in coronal section-89.23%, followed by middle section-84.19% and the apical section-64.9%. Average depth of sealer penetration for coronal section was 526.02 μm, middle-385.26 μm and apical-193.49 μm. Study concluded that there was higher epiphany sealer penetration seen in coronal followed by middle and least at apical third of the roots.

  7. Quantification of tooth wear: conventional vs new method using toolmakers microscope and a three-dimensional measuring technique.

    PubMed

    Al-Omiri, Mahmoud K; Harb, Rousan; Abu Hammad, Osama A; Lamey, Philip-John; Lynch, Edward; Clifford, Thomas J

    2010-07-01

    This study aimed to evaluate the reliability of a new CAD-CAM Laser scanning machine in detection of incisal tooth wear through a 6-month period and to compare the accuracy of using this new machine against measuring tooth wear using tool maker microscope and conventional tooth wear index. Twenty participants (11 males and 9 females, mean age=22.7 years, SD=2.0) were assessed for incisal tooth wear of lower anterior teeth using Smith and Knight clinical tooth wear index (TWI) on two occasions, the study baseline and 6 months later. Stone dies for each tooth were prepared and scanned using the CAD-CAM Laser Cercon System (Cercon Smart Ceramics, DeguDent, Germany). Scanned images were printed and examined under a toolmaker microscope (Stedall-Dowding Machine Tool Company, Optique et Mecanique de Precision, Marcel Aubert SA, Switzerland) to quantify tooth wear and then the dies were directly assessed under the microscope to measure tooth wear. The Wilcoxon Signed Ranks Test was used to analyse the data. TWI scores for incisal edges were 0, 1, and 2 and were similar at both occasions. Scores 3 and 4 were not detected. Wear values measured by directly assessing the dies under the tool maker microscope (range=517-656microm, mean=582microm, and SD=50) were significantly more than those measured from the Cercon digital machine images (range=132-193microm, mean =165microm, and SD=27) and both showed significant differences between the two occasions. Measuring images obtained with Cercon digital machine under tool maker microscope allowed detection of wear progression over the 6-month period. However, measuring the dies of worn dentition directly under the tool maker microscope enabled detection of wear progression more accurately. Conventional method was the least sensitive for tooth wear quantification and was unable to identify wear progression in most cases. Copyright 2010 Elsevier Ltd. All rights reserved.

  8. Wide field video-rate two-photon imaging by using spinning disk beam scanner

    NASA Astrophysics Data System (ADS)

    Maeda, Yasuhiro; Kurokawa, Kazuo; Ito, Yoko; Wada, Satoshi; Nakano, Akihiko

    2018-02-01

    The microscope technology with wider view field, deeper penetration depth, higher spatial resolution and higher imaging speed are required to investigate the intercellular dynamics or interactions of molecules and organs in cells or a tissue in more detail. The two-photon microscope with a near infrared (NIR) femtosecond laser is one of the technique to improve the penetration depth and spatial resolution. However, the video-rate or high-speed imaging with wide view field is difficult to perform with the conventional two-photon microscope. Because point-to-point scanning method is used in conventional one, so it's difficult to achieve video-rate imaging. In this study, we developed a two-photon microscope with spinning disk beam scanner and femtosecond NIR fiber laser with around 10 W average power for the microscope system to achieve above requirements. The laser is consisted of an oscillator based on mode-locked Yb fiber laser, a two-stage pre-amplifier, a main amplifier based on a Yb-doped photonic crystal fiber (PCF), and a pulse compressor with a pair of gratings. The laser generates a beam with maximally 10 W average power, 300 fs pulse width and 72 MHz repetition rate. And the beam incident to a spinning beam scanner (Yokogawa Electric) optimized for two-photon imaging. By using this system, we achieved to obtain the 3D images with over 1mm-penetration depth and video-rate image with 350 x 350 um view field from the root of Arabidopsis thaliana.

  9. Plasmon resonance and the imaging of metal-impregnated neurons with the laser scanning confocal microscope

    PubMed Central

    Thompson, Karen J; Harley, Cynthia M; Barthel, Grant M; Sanders, Mark A; Mesce, Karen A

    2015-01-01

    The staining of neurons with silver began in the 1800s, but until now the great resolving power of the laser scanning confocal microscope has not been utilized to capture the in-focus and three-dimensional cytoarchitecture of metal-impregnated cells. Here, we demonstrate how spectral confocal microscopy, typically reserved for fluorescent imaging, can be used to visualize metal-labeled tissues. This imaging does not involve the reflectance of metal particles, but rather the excitation of silver (or gold) nanoparticles and their putative surface plasmon resonance. To induce such resonance, silver or gold particles were excited with visible-wavelength laser lines (561 or 640 nm), and the maximal emission signal was collected at a shorter wavelength (i.e., higher energy state). Because the surface plasmon resonances of noble metal nanoparticles offer a superior optical signal and do not photobleach, our novel protocol holds enormous promise of a rebirth and further development of silver- and gold-based cell labeling protocols. DOI: http://dx.doi.org/10.7554/eLife.09388.001 PMID:26670545

  10. Alignment-free, all-spliced fiber laser source for CARS microscopy based on four-wave-mixing.

    PubMed

    Baumgartl, Martin; Gottschall, Thomas; Abreu-Afonso, Javier; Díez, Antonio; Meyer, Tobias; Dietzek, Benjamin; Rothhardt, Manfred; Popp, Jürgen; Limpert, Jens; Tünnermann, Andreas

    2012-09-10

    An environmentally-stable low-repetition rate fiber oscillator is developed to produce narrow-bandwidth pulses with several tens of picoseconds duration. Based on this oscillator an alignment-free all-fiber laser for multi-photon microscopy is realized using in-fiber frequency conversion based on four-wave-mixing. Both pump and Stokes pulses for coherent anti-Stokes Raman scattering (CARS) microscopy are readily available from one fiber end, intrinsically overlapped in space and time, which drastically simplifies the experimental handling for the user. The complete laser setup is mounted on a home-built laser scanning microscope with small footprint. High-quality multimodal microscope images of biological tissue are presented probing the CH-stretching resonance of lipids at an anti-Stokes Raman-shift of 2845 cm(-1) and second-harmonic generation of collagen. Due to its simplicity, compactness, maintenance-free operation, and ease-of-use the presented low-cost laser is an ideal source for bio-medical applications outside laser laboratories and in particular inside clinics.

  11. Experimental study of hot cracking at circular welding joints of 42CrMo steel

    NASA Astrophysics Data System (ADS)

    Zhang, Yan; Chen, Genyu; Chen, Binghua; Wang, Jinhai; Zhou, Cong

    2017-12-01

    The hot cracking at circular welding joints of quenched and tempered 42CrMo steel were studied. The flow of the molten pool and the solidification process of weld were observed with a high-speed video camera. The information on the variations in the weld temperature was collected using an infrared (IR) thermal imaging system. The metallurgical factors of hot cracking were analyzed via metallographic microscope and scanning electron microscope (SEM). The result shows that leading laser laser-metal active gas (MAG) hybrid welding process has a smaller solid-liquid boundary movement rate (VSL) and a smaller solid-liquid boundary temperature gradient (GSL) compared with leading arc laser-MAG hybrid welding process and laser welding process. Additionally, the metal in the molten pool has superior permeability while flowing toward the dendritic roots and can compensate for the inner-dendritic pressure balance. Therefore, leading laser laser-MAG hybrid welding process has the lowest hot cracking susceptibility.

  12. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: SPECTROSCOPY

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of emitted signals. The accuracy of these measurements demands that...

  13. Optical Interferometric Micrometrology

    NASA Technical Reports Server (NTRS)

    Abel, Phillip B.; Lauer, James R.

    1989-01-01

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

  14. Two-dimensional profiling of carriers in terahertz quantum cascade lasers using calibrated scanning spreading resistance microscopy and scanning capacitance microscopy.

    PubMed

    Dhar, R S; Ban, D

    2013-07-01

    The distribution of charge carriers inside the active region of a terahertz (THz) quantum cascade laser (QCL) has been measured with scanning spreading resistance microscopy (SSRM) and scanning capacitance microscopy (SCM). Individual quantum well-barrier modules with a 35.7-nm single module thickness in the active region of the device have been resolved for the first time using high-resolution SSRM and SCM techniques at room temperature. SSRM and SCM measurements on the quantum well-barrier structure were calibrated utilizing known GaAs dopant staircase samples. Doping concentrations derived from SSRM and SCM measurements were found to be in quantitative agreement with the designed average doping values of the n-type active region in the terahertz quantum cascade laser. The secondary ion mass spectroscopy provides a partial picture of internal device parameters, and we have demonstrated with our results the efficacy of uniting calibrated SSRM and SCM to delineate quantitatively the transverse cross-sectional structure of complex two-dimensional terahertz quantum cascade laser devices. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

  15. Local Resistance Profiling of Ultra Shallow Junction Annealed with Combination of Spike Lamp and Laser Annealing Processes using Scanning Spreading Resistance Microscope

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

    Abo, Satoshi; Nishikawa, Kazuhisa; Ushigome, Naoya

    2011-01-07

    Local resistance profiles of ultra shallow boron and arsenic implanted into silicon with energies of 2.0 and 4.0 keV and doses of 2.0x10{sup 15} and 1.0x10{sup 15} ions/cm{sup 2} activated by a combination of conventional spike lamp and laser annealing processes were measured by scanning spreading resistance microscope (SSRM) with a depth resolution of less than 10 nm. The lowest local resistance at the low resistance region in 2.0 keV boron implanted silicon with 1050 deg. C spike lamp annealing followed by 0.35 kW/mm{sup 2} laser annealing was half of that without laser annealing. The lowest local resistance at themore » low resistance region in the arsenic implanted silicon activated by 1050 deg. C spike lamp annealing followed by 0.39 kW/mm{sup 2} laser annealing was 74% lower than that followed by 0.36 kW/mm{sup 2} laser annealing. The lowest local resistances at the low resistance regions in the arsenic implanted silicon with 0.36 and 0.39 kW/mm{sup 2} laser annealing followed by 1050 deg. C spike lamp annealing were 41 and 33% lower than those with spike lamp annealing followed by laser annealing. Laser annealing followed by spike lamp annealing could suppress the diffusion of the impurities and was suitable for making the ultra shallow and low resistance regions.« less

  16. Ultrastructural analysis of dental ceramic surface processed by a 1070 nm fiber laser

    NASA Astrophysics Data System (ADS)

    Fornaini, C.; Merigo, E.; Poli, F.; Rocca, J.-P.; Selleri, S.; Cucinotta, A.

    2018-04-01

    Background: Lithium di-silicate dental ceramic bonding, realized by using different resins, is strictly dependent on micro-mechanical retention and chemical adhesion. The aim of this in vitro study was to investigate the capability of a 1070 nm fibre laser for their surface treatment. Methods: Samples were irradiated by a pulsed fibre laser at 1070 nm with different parameters (peak power of 5, 7.5, and 10 kW, repetition rate (RR) 20 kHz, speed of 10 and 50 mm/sec, and total energy density from 1.3 to 27 kW/cm2) Subsequently, the surface modifications were analysed by optical microscope, scanning electron microscope (SEM) and energy dispersive X-ray Spectroscopy (EDS). Results: With a peak power of 5 kW, RR of 20 kHz, and speed of 50 mm/sec, the microscopic observation of the irradiated surface showed increased roughness with small areas of melting and carbonization. EDS analysis revealed that, with these parameters, there are no evident differences between laser-processed samples and controls. Conclusions: A 1070 nm fibre laser can be considered as a good device to increase the adhesion of lithium di-silicate ceramics when optimum parameters are considered.

  17. Nano material processing with lasers in combination with nearfield technology

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

    Dickmann, K.; Jersch, J.; Demming, F.

    1996-12-31

    Recent research work has shown, that focusing of laser radiation down to a few nanometer can be obtained by using lasers in combination with nearfield technology (known from Scanning Tunneling Microscope STM or Atomic Force Microscope AFM). Lateral external illumination of STM- or AFM-probe tips with laser radiation can cause tremendous intensity enhancement in the nearfield underneath the tip. This effect can be explained by various electrostatic as well as electrodynamic effects known from Surface Enhanced Raman Spectroscopy (SERS). This effect was utilized to concentrate laser radiation with high intensity between a tip and a substrate in the nearfield. FOLANT-techniquemore » (FOcusing of LAser radiation in the Nearfield of a Tip) enables intensity enhancement up to 10{sup 6} in a narrow localized zone underneath the tip. The interaction area with nanometer scale can be applied for material processing even down to atomic dimensions. Using STM-/ laser-combination, hillocks, pits and grooves with lateral dimensions down to 10 nm have been obtained on gold substrates. AFM-/ laser-combination enabled nanostructures down to 20 nm on dielectric materials as for example polycarbonate.« less

  18. 193nm high power lasers for the wide bandgap material processing

    NASA Astrophysics Data System (ADS)

    Fujimoto, Junichi; Kobayashi, Masakazu; Kakizaki, Koji; Oizumi, Hiroaki; Mimura, Toshio; Matsunaga, Takashi; Mizoguchi, Hakaru

    2017-02-01

    Recently infrared laser has faced resolution limit of finer micromachining requirement on especially semiconductor packaging like Fan-Out Wafer Level Package (FO-WLP) and Through Glass Via hole (TGV) which are hard to process with less defect. In this study, we investigated ablation rate with deep ultra violet excimer laser to explore its possibilities of micromachining on organic and glass interposers. These results were observed with a laser microscopy and Scanning Electron Microscope (SEM). As the ablation rates of both materials were quite affordable value, excimer laser is expected to be put in practical use for mass production.

  19. Mechanical design of a precision linear flexural stage for 3D x-ray diffraction microscope at the Advanced Photon Source

    NASA Astrophysics Data System (ADS)

    Shu, D.; Liu, W.; Kearney, S.; Anton, J.; Tischler, J. Z.

    2015-09-01

    The 3-D X-ray diffraction microscope is a new nondestructive tool for the three-dimensional characterization of mesoscopic materials structure. A flexural-pivot-based precision linear stage has been designed to perform a wire scan as a differential aperture for the 3-D diffraction microscope at the Advanced Photon Source, Argonne National Laboratory. The mechanical design and finite element analyses of the flexural stage, as well as its initial mechanical test results with laser interferometer are described in this paper.

  20. Direct-write maskless lithography using patterned oxidation of Si-substrate Induced by femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Kiani, Amirkianoosh; Venkatakrishnan, Krishnan; Tan, Bo

    2013-03-01

    In this study we report a new method for direct-write maskless lithography using oxidized silicon layer induced by high repetition (MHz) ultrafast (femtosecond) laser pulses under ambient condition. The induced thin layer of predetermined pattern can act as an etch stop during etching process in alkaline etchants such as KOH. The proposed method can be leading to promising solutions for direct-write maskless lithography technique since the proposed method offers a higher degree of flexibility and reduced time and cost of fabrication which makes it particularly appropriate for rapid prototyping and custom scale manufacturing. A Scanning Electron Microscope (SEM), Micro-Raman, Energy Dispersive X-ray (EDX), optical microscope and X-ray diffraction spectroscopy (XRD) were used to evaluate the quality of oxidized layer induced by laser pulses.

  1. Development and applications of optical interferometric micrometrology in the angstrom and subangstrom range

    NASA Technical Reports Server (NTRS)

    Lauer, James L.; Abel, Phillip B.

    1988-01-01

    The recent development of the scanning electron tunneling microscope and the atomic force microscope requires absolute standards for measurements in the angstrom and subangstrom range. Optical interferometry with lasers and multiple mode laser resonances can provide absolute measurements as the laser wavelengths are very accurately known. A key feature of such measurements is the use of piezoelectric crystals as translators of the highest accuracy for very small disturbances. However, the dimensional changes of these crystals resulting from electrical potential changes depend on many variables, among them the method of mounting, so that accurate calibrations are necessary. Starting from advances in optical metrology made by physicists trying to find gravity waves, advances which led to measurements down to 10 to the -5 A, the author designed and built a much simpler system for the angstrom range. The major limiting factors were mechanical vibrations, air currents, thermal changes and laser instabilities.

  2. Large scale infrared imaging of tissue micro arrays (TMAs) using a tunable Quantum Cascade Laser (QCL) based microscope.

    PubMed

    Bassan, Paul; Weida, Miles J; Rowlette, Jeremy; Gardner, Peter

    2014-08-21

    Chemical imaging in the field of vibrational spectroscopy is developing into a promising tool to complement digital histopathology. Applications include screening of biopsy tissue via automated recognition of tissue/cell type and disease state based on the chemical information from the spectrum. For integration into clinical practice, data acquisition needs to be speeded up to implement a rack based system where specimens are rapidly imaged to compete with current visible scanners where 100's of slides can be scanned overnight. Current Fourier transform infrared (FTIR) imaging with focal plane array (FPA) detectors are currently the state-of-the-art instrumentation for infrared absorption chemical imaging, however recent development in broadly tunable lasers in the mid-IR range is considered the most promising potential candidate for next generation microscopes. In this paper we test a prototype quantum cascade laser (QCL) based spectral imaging microscope with a focus on discrete frequency chemical imaging. We demonstrate how a protein chemical image of the amide I band (1655 cm(-1)) of a 2 × 2.4 cm(2) breast tissue microarray (TMA) containing over 200 cores can be measured in 9 min. This result indicates that applications requiring chemical images from a few key wavelengths would be ideally served by laser-based microscopes.

  3. Design and installation of a multimode microscopy system

    NASA Astrophysics Data System (ADS)

    Helm, Johannes P.; Haug, Finn-Mogens S.; Storm, Johan F.; Ottersen, Ole-Petter

    2001-04-01

    We describe design and installation of a multi-mode microscopy core facility in an environment of varied research activity in life-sciences. The experimentators can select any combination of a) microscopes (upright, upright fixed-stage, inverted), b) microscopy modes (widefield, DIC, IRDIC, widefield epifluorescence, transmission LSM, reflection and fluorescence CLSM, MPLSM), c) imaging techniques (direct observation, video observation, photography, quantitative camera-recording, flying spot scanning), d) auxiliary systems (equipment for live specimen imaging, electrophysiology, time-coordinated laser-scanning and electrophysiology, patch-clamp). The equipment is installed on one large vibration-isolating optical table (3m X 1.5m X 0.3m). Electronics, auxiliary equipment, and a fiber-coupled, remotely controlled Ar+-Kr+ laser are mounted in a rack system fixed to the ceiling. The design of the shelves allows the head of the CSLM to be moved to any of the microscopes without increasing critical cable lengths. At the same time easy access to all the units is preserved. The beam of a Titanium-Sapphire laser, controlled by means of an EOM and a prism GVD, is coupled directly to the microscopes. Three mirrors mounted on a single precision translation table are integrated into the beam steering system so that the beam can easily be redirected to any of the microscopes. All the available instruments can be operated by the educated and trained user. The system is popular among researchers in neuroanatomy, embryology, cell biology, molecular biology - including the study of protein interactions, e.g. by means of FRET, and electrophysiology. Its colocalization with an EM facility promises to provide considerable synergy effects.

  4. Comparative scanning electron microscope analysis of diode laser and desensitizing toothpastes for evaluation of efficacy of dentinal tubular occlusion

    PubMed Central

    Reddy, Guntakala Vikram; Akula, Sushma; Malgikar, Suryakanth; Babu, Palaparthy Raja; Reddy, Gooty Jagadish; Josephin, Johnson Juliet

    2017-01-01

    Background: The present study aims to evaluate the efficacy of diode laser alone and in combination with desensitizing toothpastes in occluding dentinal tubules (both partially occluded and completely occluded tubules) by scanning electron microscope (SEM). Materials and Methods: Fifty human teeth were extracted, cervical cavities were prepared and etched with 17% ethylenediaminetetraacetic acid, and smear layer was removed to expose the tubules. The teeth were divided into five groups: Group I – Application of NovaMin-formulated toothpaste, Group II – Application of Pro-Argin™-formulated toothpaste, Group III – Application of diode laser in noncontact mode, Group IV – NovaMin-formulated toothpaste followed by laser irradiation, and Group V – Pro-Argin™-formulated toothpaste followed by laser irradiation. After treatment, quantitative analysis of occluded dentinal tubules was done by SEM analysis. Results: The mean values of percentages of total occlusion of dentinal tubules in Groups I, II, III, IV, and V were 92.73% ± 1.38, 90.67% ± 1.86, 96.57% ± 0.64, 97.3% ± 0.68, and 96.9% ± 6.08, respectively. Addition of diode laser (Groups III, IV, and V) yielded a significant occlusion of the dentinal tubules when compared to desensitizing toothpastes alone (Groups I and II). Conclusion: Diode laser (Group III) has shown more efficacy in occluding dentinal tubules when compared with desensitizing toothpastes which was statistically significant (P < 0.05). Among the five groups, NovaMin + diode laser (Group IV) showed the highest percentage of occluded dentinal tubules. PMID:29398853

  5. A 3D Polymer Based Printed Two-Dimensional Laser Scanner

    NASA Astrophysics Data System (ADS)

    Oyman, H. A.; Gokdel, Y. D.; Ferhanoglu, O.; Yalcinkaya, A. D.

    2016-10-01

    A two-dimensional (2D) polymer based scanning mirror with magnetic actuation is developed for imaging applications. Proposed device consists of a circular suspension holding a rectangular mirror and can generate a 2D scan pattern. Three dimensional (3D) printing technology which is used for implementation of the device, offers added flexibility in controlling the cross-sectional profile as well as the stress distribution compared to the traditional planar process technologies. The mirror device is developed to meet a portable, miniaturized confocal microscope application in mind, delivering 4.5 and 4.8 degrees of optical scan angles at 111 and 267 Hz, respectively. As a result of this mechanical performance, the resulting microscope incorporating the mirror is estimated to accomplish a field of view (FOV) of 350 µm × 350 µm.

  6. Realistic representation of Bacillus subtilis biofilms architecture using combined microscopy (CLSM, ESEM and FESEM).

    PubMed

    Bridier, A; Meylheuc, T; Briandet, R

    2013-05-01

    In this contribution, we used a set of microscopic techniques including confocal laser scanning microscopy (CLSM), environmental scanning electron microscopy (ESEM) and field emission scanning electron microscopy (FESEM) to analyze the three-dimensional spatial arrangement of cells and their surrounding matrix in Bacillus subtilis biofilm. The combination of the different techniques enabled a deeper and realistic deciphering of biofilm architecture by providing the opportunity to overcome the limits of each single technique. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Microanalysis of tool steel and glass with laser-induced breakdown spectroscopy

    NASA Astrophysics Data System (ADS)

    Loebe, Klaus; Uhl, Arnold; Lucht, Hartmut

    2003-10-01

    A laser microscope system for the microanalytical characterization of complex materials is described. The universal measuring principle of laser-induced breakdown spectroscopy (LIBS) in combination with echelle optics permits a fast simultaneous multielement analysis with a possible spatial resolution below 10 pm. The developed system features completely UV-transparent optics for the laser-microscope coupling and the emission beam path and enables parallel signal detection within the wavelength range of 200-800 nm with a spectral resolution of a few picometers. Investigations of glass defects and tool steels were performed. The characterization of a glass defect in a tumbler by a micro-LIBS line scan, with use of a 266-nm diode-pumped Nd:YAG laser for excitation, is possible by simple comparison of plasma spectra of the defect and the surrounding area. Variations in the main elemental composition as well as impurities by trace elements are detected at the same time. Through measurement of the calibration samples with the known concentration of the corresponding element, a correlation between the intensity of spectral lines and the element concentration was also achieved. The change of elemental composition at the transient stellite solder of tool steels has been determined by an area scan. The two-dimensional pictures show abrupt changes of the element distribution along the solder edge and allow fundamental researches of dynamic modifications (e.g., diffusion) in steel.

  8. A diamond-based scanning probe spin sensor operating at low temperature in ultra-high vacuum

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

    Schaefer-Nolte, E.; Wrachtrup, J.; 3rd Institute of Physics and Research Center SCoPE, University Stuttgart, 70569 Stuttgart

    2014-01-15

    We present the design and performance of an ultra-high vacuum (UHV) low temperature scanning probe microscope employing the nitrogen-vacancy color center in diamond as an ultrasensitive magnetic field sensor. Using this center as an atomic-size scanning probe has enabled imaging of nanoscale magnetic fields and single spins under ambient conditions. In this article we describe an experimental setup to operate this sensor in a cryogenic UHV environment. This will extend the applicability to a variety of molecular systems due to the enhanced target spin lifetimes at low temperature and the controlled sample preparation under UHV conditions. The instrument combines amore » tuning-fork based atomic force microscope (AFM) with a high numeric aperture confocal microscope and the facilities for application of radio-frequency (RF) fields for spin manipulation. We verify a sample temperature of <50 K even for strong laser and RF excitation and demonstrate magnetic resonance imaging with a magnetic AFM tip.« less

  9. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: FOUNDATIONS FOR CALIBRATION, QUANTITATION AND SPECTROSCOPY

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The goal of a CLSM is to acquire and quantify fluorescence and in some instruments acquire spectral characterization of emitted signals. The accuracy of these measurements demands that...

  10. Studies of porphyrin-containing specimens using an optical spectrometer connected to a confocal scanning laser microscope.

    PubMed

    Trepte, O; Rokahr, I; Andersson-Engels, S; Carlsson, K

    1994-12-01

    A spectrometer has been developed for use with a confocal scanning laser microscope. With this unit, spectral information from a single point or a user-defined region within the microscope specimen can be recorded. A glass prism is used to disperse the spectral components of the recorded light over a linear CCD photodiode array with 256 elements. A regulated cooling unit keeps the detector at 277 K, thereby allowing integration times of up to 60 s. The spectral resolving power, lambda/delta lambda, ranges from 350 at lambda = 400 nm to 100 at lambda = 700 nm. Since the entrance aperture of the spectrometer has the same size as the detector pinhole used during normal confocal scanning, the three-dimensional spatial resolution is equivalent to that of normal confocal scanning. Light from the specimen is deflected to the spectrometer by a solenoid controlled mirror, allowing fast and easy switching between normal confocal scanning and spectrometer readings. With this equipment, studies of rodent liver specimens containing porphyrins have been made. The subcellular localization is of interest for the mechanisms of photodynamic therapy (PDT) of malignant tumours. Spectroscopic detection is necessary to distinguish the porphyrin signal from other fluorescent components in the specimen. Two different substances were administered to the tissue, Photofrin, a haematoporphyrin derivative (HPD) and delta-amino levulinic acid (ALA), a precursor to protoporphyrin IX and haem in the haem cycle. Both are substances under clinical trials for PDT of malignant tumours. Following administration of these compounds to the tissue, the potent photosensitizer and fluorescent compound Photofrin, or protoporphyrin IX, respectively, is accumulated.(ABSTRACT TRUNCATED AT 250 WORDS)

  11. Lattice relations and solidification of the complex regular eutectic (Cr,Fe)-(Cr,Fe)23C6

    NASA Astrophysics Data System (ADS)

    Lai, Hsuan-Han; Hsieh, Chih-Chun; Lin, Chi-Ming; Wu, Weite

    2017-05-01

    The eutectic (Cr,Fe)-(Cr,Fe)23C6 showed a triaxial fishbone structure and could be categorized as a "complex regular structure". In this study, the lattice relations of the fishbone (Cr,Fe)23C6 were examined and the solidification process was observed using a transmission electron microscope and a confocal laser scanning microscope. For one of the three fish bones in a eutectic cell, parallel (Cr,Fe)23C6 lamellas at one side of the spine had the same lattice direction, as did those in the (Cr,Fe) phase. The lattices of neighboring (Cr,Fe)23C6 and (Cr,Fe) phases were not coherent. Lamellar (Cr,Fe)23C6 on opposite sides of a spine had different lattice directions, and their lattice boundary was in the spine. By using the confocal laser scanning microscope, the solidification of lamellar eutectic structure could be observed. At the low cooling rate of 5 o C·min-1, parallel lamellas would grow thick blocks instead of thin plates. To obtain a thin lamellar eutectic structure, the cooling rate should be higher, like the rate in welding.

  12. Granulocytes of the red claw crayfish Cherax quadricarinatus can endocytose beads, E. coli and WSSV, but in different ways.

    PubMed

    Duan, Hu; Jin, Songjun; Zhang, Yan; Li, Fuhua; Xiang, Jianhai

    2014-10-01

    The hemocytes of the red claw crayfish Cherax quadricarinatus are classified by morphologic observation into the following types: hyalinocytes (H), semi-granulocytes (SG) and granulocytes (G). Density gradient centrifugation with Percoll was developed to separate these three subpopulations of hemocytes. Beads, Escherichia coli, and FITC labeling WSSV were used to investigate the characteristics of granulocytes by using scanning electron microscope, transmission electron microscope, and laser scan confocal microscope. Results showed that granulocytes could phagocytose beads and E. coli by endocytic pathways. WSSV could rely on caveolae-mediated endocytosis to mainly enter into granulocytes. These results could elucidate the mechanism of the innate immunity function of granulocytes, and it also showed the mechanism by which WSSV invaded granulocytes in the red claw crayfish. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Development of a Hybrid Atomic Force Microscopic Measurement System Combined with White Light Scanning Interferometry

    PubMed Central

    Guo, Tong; Wang, Siming; Dorantes-Gonzalez, Dante J.; Chen, Jinping; Fu, Xing; Hu, Xiaotang

    2012-01-01

    A hybrid atomic force microscopic (AFM) measurement system combined with white light scanning interferometry for micro/nanometer dimensional measurement is developed. The system is based on a high precision large-range positioning platform with nanometer accuracy on which a white light scanning interferometric module and an AFM head are built. A compact AFM head is developed using a self-sensing tuning fork probe. The head need no external optical sensors to detect the deflection of the cantilever, which saves room on the head, and it can be directly fixed under an optical microscopic interferometric system. To enhance the system’s dynamic response, the frequency modulation (FM) mode is adopted for the AFM head. The measuring data can be traceable through three laser interferometers in the system. The lateral scanning range can reach 25 mm × 25 mm by using a large-range positioning platform. A hybrid method combining AFM and white light scanning interferometry is proposed to improve the AFM measurement efficiency. In this method, the sample is measured firstly by white light scanning interferometry to get an overall coarse morphology, and then, further measured with higher resolution by AFM. Several measuring experiments on standard samples demonstrate the system’s good measurement performance and feasibility of the hybrid measurement method. PMID:22368463

  14. Development of a hybrid atomic force microscopic measurement system combined with white light scanning interferometry.

    PubMed

    Guo, Tong; Wang, Siming; Dorantes-Gonzalez, Dante J; Chen, Jinping; Fu, Xing; Hu, Xiaotang

    2012-01-01

    A hybrid atomic force microscopic (AFM) measurement system combined with white light scanning interferometry for micro/nanometer dimensional measurement is developed. The system is based on a high precision large-range positioning platform with nanometer accuracy on which a white light scanning interferometric module and an AFM head are built. A compact AFM head is developed using a self-sensing tuning fork probe. The head need no external optical sensors to detect the deflection of the cantilever, which saves room on the head, and it can be directly fixed under an optical microscopic interferometric system. To enhance the system's dynamic response, the frequency modulation (FM) mode is adopted for the AFM head. The measuring data can be traceable through three laser interferometers in the system. The lateral scanning range can reach 25 mm × 25 mm by using a large-range positioning platform. A hybrid method combining AFM and white light scanning interferometry is proposed to improve the AFM measurement efficiency. In this method, the sample is measured firstly by white light scanning interferometry to get an overall coarse morphology, and then, further measured with higher resolution by AFM. Several measuring experiments on standard samples demonstrate the system's good measurement performance and feasibility of the hybrid measurement method.

  15. Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope

    NASA Astrophysics Data System (ADS)

    Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

    2016-03-01

    Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

  16. Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope.

    PubMed

    Batista, Ana; Breunig, Hans Georg; Uchugonova, Aisada; Morgado, António Miguel; König, Karsten

    2016-03-01

    Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

  17. Corneal reshaping using a pulsed UV solid-state laser

    NASA Astrophysics Data System (ADS)

    Ren, Qiushi; Simon, Gabriel; Parel, Jean-Marie A.; Shen, Jin-Hui; Takesue, Yoshiko

    1993-06-01

    Replacing the gas ArF (193 nm) excimer laser with a solid state laser source in the far-UV spectrum region would eliminate the hazards of a gas laser and would reduce its size which is desirable for photo-refractive keratectomy (PRK). In this study, we investigated corneal reshaping using a frequency-quintupled (213 nm) pulsed (10 ns) Nd:YAG laser coupled to a computer-controlled optical scanning delivery system. Corneal topographic measurements showed myopic corrections ranging from 2.3 to 6.1 diopters. Post-operative examination with the slit-lamp and operating microscope demonstrated a smoothly ablated surface without corneal haze. Histological results showed a smoothly sloping surface without recognizable steps. The surface quality and cellular effects were similar to that of previously described excimer PRK. Our study demonstrated that a UV solid state laser coupled to an optical scanning delivery system is capable of reshaping the corneal surface with the advantage of producing customized, aspheric corrections without corneal haze which may improve the quality of vision following PRK.

  18. Three-dimensional non-destructive optical evaluation of laser-processing performance using optical coherence tomography.

    PubMed

    Kim, Youngseop; Choi, Eun Seo; Kwak, Wooseop; Shin, Yongjin; Jung, Woonggyu; Ahn, Yeh-Chan; Chen, Zhongping

    2008-06-01

    We demonstrate the use of optical coherence tomography (OCT) as a non-destructive diagnostic tool for evaluating laser-processing performance by imaging the features of a pit and a rim. A pit formed on a material at different laser-processing conditions is imaged using both a conventional scanning electron microscope (SEM) and OCT. Then using corresponding images, the geometrical characteristics of the pit are analyzed and compared. From the results, we could verify the feasibility and the potential of the application of OCT to the monitoring of the laser-processing performance.

  19. Investigation of laser irradiation of WC-Co cemented carbides inside a scanning electron microscope (LASEM)

    NASA Astrophysics Data System (ADS)

    Schultrich, B.; Wetzig, K.

    1987-09-01

    A combination of SEM and laser enables direct observation of structural modifications by a high-energy input. With this new device, melting phenomena and fracture processes in a WC-6 percent Co hard metal were investigated. The first laser pulse leads to melting of a thin surface layer with the formation of blisters and craters. Cracking is induced by the relaxation of compressive surface stresses during the high-temperature stage and the appearance of tensile stresses during cooling. Besides crack formation and extension, complete welding of crack surfaces was observed after repeated laser irradiation.

  20. Three-dimensional non-destructive optical evaluation of laser-processing performance using optical coherence tomography

    PubMed Central

    Kim, Youngseop; Choi, Eun Seo; Kwak, Wooseop; Shin, Yongjin; Jung, Woonggyu; Ahn, Yeh-Chan; Chen, Zhongping

    2014-01-01

    We demonstrate the use of optical coherence tomography (OCT) as a non-destructive diagnostic tool for evaluating laser-processing performance by imaging the features of a pit and a rim. A pit formed on a material at different laser-processing conditions is imaged using both a conventional scanning electron microscope (SEM) and OCT. Then using corresponding images, the geometrical characteristics of the pit are analyzed and compared. From the results, we could verify the feasibility and the potential of the application of OCT to the monitoring of the laser-processing performance. PMID:24932051

  1. Scanning electron microscope and dye penetration test: comparison of root canal preparation with 15 F CO2 laser microprobe versus conventional method--in vivo study

    NASA Astrophysics Data System (ADS)

    Kesler, Gavriel; Koren, Rumelia; Kesler, Anat; Hay, Nissim; Gal, Rivka

    1999-05-01

    The study was conducted on 30 vital maxillary or mandibulary teeth destined for extraction due to periodontal problems. 21 were experimentally treated with pulsed CO2 laser delivered by a newly developed fiber and 9 teeth represented the control group. The micro probe is a flexible, hollow, metal fiber, 300 μm in diameter and 20 mm in length, coupled onto a handpiece, with the following radiation parameters: wavelength-10.6μm pulse duration-50m.sec; energy per pulses 0.25 joule; energy density-360 J/cm2 per pulse; power on tissue-5W. The laser group was divided into three, receiving 20, 40 or 60 pulses, respectively. On light microscopy: in all the control group cases, large amount of residual pulp tissue was seen, it was diminished in some of the low energy group and was totally eradicated in the high energy group. This was confirmed by the scanning electron microscope (SEM) examination. The dentin tubuli were partly occluded with the low energy levels and completely with the high levels, as shown by the high-speed centrifuge dye penetration test and by the SEM tests.

  2. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

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

    Darvin, M E; Richter, H; Zhu, Y J

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed bymore » using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted. (laser biophotonics)« less

  3. SU-E-T-98: Towards Cell Nucleus Microdosimetry: Construction of a Confocal Laser-Scanning Fluorescence Microscope to Readout Fluorescence Nuclear Track Detectors (FNTDs).

    PubMed

    McFadden, C; Bartz, J; Akselrod, M; Sawakuchi, G

    2012-06-01

    To construct a custom confocal laser scanning microscope (CLSM) capable of resolving individual proton tracks in the volume of an Al 2 O 3 :C,Mg fluorescent nuclear track detector (FNTD). The spatial resolution of the FNTD technique is at the sub-micrometer scale. Therefore the FNTD technique has the potential to perform radiation measurements at the cell nucleus scale. The crystal volume of an FNTD contains defects which become fluorescent F 2 + centers after trapping delta electrons from ionizing radiation. These centers have an absorption band centered at 620 nm and an emission band in the near infrared. Events of energy deposition in the crystal are read-out using a CLSM with sub-micrometer spatial resolution. Excitation light from a 635 nm laser is focused in the crystal volume by an objective lens. Fluorescence is collected back through the same path, filtered through a dichroic mirror, and focused through a small pinhole onto an avalanche photodiode. Lateral scanning of the focal point is performed with a scanning mirror galvanometer, and axial scanning is performed using a stepper-motor stage. Control of electronics and image acquisition was performed using a custom built LabVIEW VI and further image processing was done using Java. The system was used to scan FNTDs exposed to a 6 MV x-ray beam and an unexposed FNTD. Fluorescence images above the unexposed background were obtained at scan depths ranging from 5 - 10 micrometer below the crystal surface using a 100 micrometer pinhole size. Further work needs to be done to increase the resolution and the signal to noise ratio of the images so that energy deposition events may be identified more easily. Natural Sciences and Engineering Research Council of Canada. © 2012 American Association of Physicists in Medicine.

  4. Widely tuneable scattering-type scanning near-field optical microscopy using pulsed quantum cascade lasers

    NASA Astrophysics Data System (ADS)

    Yoxall, Edward; Navarro-Cía, Miguel; Rahmani, Mohsen; Maier, Stefan A.; Phillips, Chris C.

    2013-11-01

    We demonstrate the use of a pulsed quantum cascade laser, wavelength tuneable between 6 and 10 μm, with a scattering-type scanning near-field optical microscope (s-SNOM). A simple method for calculating the signal-to-noise ratio (SNR) of the s-SNOM measurement is presented. For pulsed lasers, the SNR is shown to be highly dependent on the degree of synchronization between the laser pulse and the sampling circuitry; in measurements on a gold sample, the SNR is 26 with good synchronization and less than 1 without. Simulations and experimental s-SNOM images, with a resolution of 100 nm, corresponding to λ/80, and an acquisition time of less than 90 s, are presented as proof of concept. They show the change in the field profile of plasmon-resonant broadband antennas when they are excited with wavelengths of 7.9 and 9.5 μm.

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

    Henn, T.; Kiessling, T., E-mail: tobias.kiessling@physik.uni-wuerzburg.de; Ossau, W.

    We describe a two-color pump-probe scanning magneto-optical Kerr effect microscope which we have developed to investigate electron spin phenomena in semiconductors at cryogenic temperatures with picosecond time and micrometer spatial resolution. The key innovation of our microscope is the usage of an ultrafast “white light” supercontinuum fiber-laser source which provides access to the whole visible and near-infrared spectral range. Our Kerr microscope allows for the independent selection of the excitation and detection energy while avoiding the necessity to synchronize the pulse trains of two separate picosecond laser systems. The ability to independently tune the pump and probe wavelength enables themore » investigation of the influence of excitation energy on the optically induced electron spin dynamics in semiconductors. We demonstrate picosecond real-space imaging of the diffusive expansion of optically excited electron spin packets in a (110) GaAs quantum well sample to illustrate the capabilities of the instrument.« less

  6. Laser ablation of Au-CuO core-shell nanocomposite in water for optoelectronic devices

    NASA Astrophysics Data System (ADS)

    Ismail, Raid A.; Abdul-Hamed, Ryam S.

    2017-12-01

    Core-shell gold-copper oxide Au-CuO nanocomposites were synthesized using laser ablation of CuO target in colloidal solution of Au nanoparticles (NPs). The effect of laser fluence on the structural, morphological, electrical, and optical properties of Au-CuO nanocomposites was investigated using x-ray diffraction (XRD), atomic force microscope (AFM), scanning electron microscope (SEM), transmission electron microscope (TEM), photoluminescence (PL), Fourier transformed infrared spectroscopy (FTIR), Hall measurement, and UV-vis spectroscopy. X-ray diffraction results confirm the formation of polycrystalline Au-CuO NPs with monoclinic structure. The optical energy gap for CuO was 4 eV and for the Au-CuO core-shell nanocomposites was found to be in the range of 3.4-3.7 eV. SEM and TEM investigations revealed that the structure and morphology of Au-CuO core-shell nanocomposites were strongly depending on the laser fluence. A formation of Au-CuO nanospheres and platelets structures was observed. The photoluminescence data showed an emission of broad visible peaks between 407 and 420 nm. The effect of laser fluence on the dark and illuminated I-V characteristics of Au-CuO/n-Si heterojunction photodetectors was investigated and analyzed. The experimental data demonstrated that the photodetector prepared at optimum laser fluence exhibited photosensitivity of 0.6 AW-1 at 800 nm.

  7. Video-rate volumetric functional imaging of the brain at synaptic resolution.

    PubMed

    Lu, Rongwen; Sun, Wenzhi; Liang, Yajie; Kerlin, Aaron; Bierfeld, Jens; Seelig, Johannes D; Wilson, Daniel E; Scholl, Benjamin; Mohar, Boaz; Tanimoto, Masashi; Koyama, Minoru; Fitzpatrick, David; Orger, Michael B; Ji, Na

    2017-04-01

    Neurons and neural networks often extend hundreds of micrometers in three dimensions. Capturing the calcium transients associated with their activity requires volume imaging methods with subsecond temporal resolution. Such speed is a challenge for conventional two-photon laser-scanning microscopy, because it depends on serial focal scanning in 3D and indicators with limited brightness. Here we present an optical module that is easily integrated into standard two-photon laser-scanning microscopes to generate an axially elongated Bessel focus, which when scanned in 2D turns frame rate into volume rate. We demonstrated the power of this approach in enabling discoveries for neurobiology by imaging the calcium dynamics of volumes of neurons and synapses in fruit flies, zebrafish larvae, mice and ferrets in vivo. Calcium signals in objects as small as dendritic spines could be resolved at video rates, provided that the samples were sparsely labeled to limit overlap in their axially projected images.

  8. Carbon dioxide laser microsurgery of the uterine tube.

    PubMed

    Baggish, M S; Chong, A P

    1981-07-01

    The carbon dioxide (CO2) laser was used to perform microsurgical excision of obstructed tubal segments in rabbit and human subjects. Approximation of the freshly severed tubes by means of laser "welding" was evaluated in both groups investigated. More important, the laser beam cuts accurately and atraumatically while sealing small vascular channels. Scanning electron microscopic studies of the human fallopian tube following laser surgery were done to determine the extent of tissue injury. At a distance of 1 mm distal to the vaporization and necrotic impact zone, normal tubal anatomy was observed. Follow-up data are presented for 7 women who underwent laser beam tuboplasty between 1979 and 1980. The principle advantages of the CO2 laser are its precise control, minimal tissue injury, and hemostatic properties.

  9. High contrast imaging and flexible photomanipulation for quantitative in vivo multiphoton imaging with polygon scanning microscope.

    PubMed

    Li, Yongxiao; Montague, Samantha J; Brüstle, Anne; He, Xuefei; Gillespie, Cathy; Gaus, Katharina; Gardiner, Elizabeth E; Lee, Woei Ming

    2018-02-28

    In this study, we introduce two key improvements that overcome limitations of existing polygon scanning microscopes while maintaining high spatial and temporal imaging resolution over large field of view (FOV). First, we proposed a simple and straightforward means to control the scanning angle of the polygon mirror to carry out photomanipulation without resorting to high speed optical modulators. Second, we devised a flexible data sampling method directly leading to higher image contrast by over 2-fold and digital images with 100 megapixels (10 240 × 10 240) per frame at 0.25 Hz. This generates sub-diffraction limited pixels (60 nm per pixels over the FOV of 512 μm) which increases the degrees of freedom to extract signals computationally. The unique combined optical and digital control recorded fine fluorescence recovery after localized photobleaching (r ~10 μm) within fluorescent giant unilamellar vesicles and micro-vascular dynamics after laser-induced injury during thrombus formation in vivo. These new improvements expand the quantitative biological-imaging capacity of any polygon scanning microscope system. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. TOPSIS based parametric optimization of laser micro-drilling of TBC coated nickel based superalloy

    NASA Astrophysics Data System (ADS)

    Parthiban, K.; Duraiselvam, Muthukannan; Manivannan, R.

    2018-06-01

    The technique for order of preference by similarity ideal solution (TOPSIS) approach was used for optimizing the process parameters of laser micro-drilling of nickel superalloy C263 with Thermal Barrier Coating (TBC). Plasma spraying was used to deposit the TBC and a pico-second Nd:YAG pulsed laser was used to drill the specimens. Drilling angle, laser scan speed and number of passes were considered as input parameters. Based on the machining conditions, Taguchi L8 orthogonal array was used for conducting the experimental runs. The surface roughness and surface crack density (SCD) were considered as the output measures. The surface roughness was measured using 3D White Light Interferometer (WLI) and the crack density was measured using Scanning Electron Microscope (SEM). The optimized result achieved from this approach suggests reduced surface roughness and surface crack density. The holes drilled at an inclination angle of 45°, laser scan speed of 3 mm/s and 400 number of passes found to be optimum. From the Analysis of variance (ANOVA), inclination angle and number of passes were identified as the major influencing parameter. The optimized parameter combination exhibited a 19% improvement in surface finish and 12% reduction in SCD.

  11. Temperature induced degradation mechanisms of AlInAs/InGaAs/InP quantum cascade lasers

    NASA Astrophysics Data System (ADS)

    Pierścińska, D.; Pierściński, K.; Płuska, M.; Sobczak, G.; Kuźmicz, A.; Gutowski, P.; Bugajski, M.

    2018-01-01

    In this paper, we report on the investigation of temperature induced degradation mode of quantum cascade lasers (QCLs) with an emphasis on the influence of different processing technology. We investigate and compare lattice matched AlInAs/InGaAs/InP QCLs of various constructions, i.e., double trench, buried heterostructure and ridge waveguide regarding thermal management, reliability and sources of degradation. The analysis was performed by CCD thermoreflectance spectroscopy, scanning electron microscope inspection and destructive analysis by focused ion beam etching, enabling determination of the source and mode of degradation for investigated lasers. Experimental temperature data relate temperature rise, arising from supply current, with device geometry. Results clearly indicate, that the buried heterostructure geometry, allows reaching the highest maximal operating current densities, before the degradation occurs. Microscopic images of degradation confirm that degradation includes the damage of the contact layer as well as damage of the active region layers.

  12. SEM investigations of the cementum surface after irradiation with a frequency-doubled Alexandrite laser

    NASA Astrophysics Data System (ADS)

    Rechmann, Peter; Hennig, Thomas

    1996-04-01

    During prior studies it could be demonstrated while engaging a frequency doubled Alexandrite-laser (wavelength 380 nm, pulse duration 100 ns, fluence 1 J/cm2, pulse repetition rate 110 Hz) a fast and strictly selective ablation of supra- and subgingival calculus is possible. Even the removal of unstained microbial plaque was observed. First conclusions were drawn after light microscopical investigations on undecalcified sections of irradiated teeth. In the present study the cementum surface after irradiation with a frequency doubled Alexandrite-laser was observed by means of a Scanning Electron Microscope. After irradiation sections of teeth were dried in alcohol and sputtered with gold. In comparison irradiated cementum surfaces of unerupted operatively removed wisdom teeth and tooth surfaces after the selective removal of calculus were investigated. A complete removal of calculus was observed as well as a remaining smooth surface of irradiated cementum.

  13. Electron Beam/Laser Glazing of Iron-Base Materials.

    DTIC Science & Technology

    1981-07-01

    alloy (-l. 5wt %Cr) steels after laser and electron beam glazing. In this work it is shown that the dramatic difference in microstructure and hardness...highly alloyed tool steels can be critical in determining the complexity of the solidification route. The analyses of M2, M42 and M7 are given in...the type described in Fe- Ni alloys (1). This con- clusion is based on optical and scanning electron microscope observation unambig- uously showing

  14. Evaluation of Wear on Macro-Surface Textures Generated by ns Fiber Laser

    NASA Astrophysics Data System (ADS)

    Harish, V.; Soundarapandian, S.; Vijayaraghavan, L.; Bharatish, A.

    2018-03-01

    The demand for improved performance and long term reliability of mechanical systems dictate the use of advanced materials and surface engineering techniques. A small change in the surface topography can lead to substantial improvements in the tribological behaviour of the contact surfaces. One way of altering the surface topography is by surface texturing by introducing dimples or channels on the surfaces. Surface texturing is already a successful technique which finds a wide area of applications ranging from heavy industries to small scale devices. This paper reports the effect of macro texture shapes generated using a nanosecond fiber laser on wear of high carbon chromium steel used in large size bearings having rolling contacts. Circular and square shaped dimples were generated on the surface to assess the effect of sliding velocities on friction coefficient. Graphite was used as solid lubricant to minimise the effect of wear on textured surfaces. The laser parameters such as power, scan speed and passes were optimised to obtain macro circular and square dimples which was characterised using a laser confocal microscope. The friction coefficients of the circular and square dimples were observed to lie in the same range due to minimum wear on the surface. On the contrary, at medium and higher sliding velocities, square dimples exhibited lower friction coefficient values compared to circular dimples. The morphology of textured specimen was characterised using Scanning Electron Microscope.

  15. Optical toolkits for in vivo deep tissue laser scanning microscopy: a primer

    NASA Astrophysics Data System (ADS)

    Lee, Woei Ming; McMenamin, Thomas; Li, Yongxiao

    2018-06-01

    Life at the microscale is animated and multifaceted. The impact of dynamic in vivo microscopy in small animals has opened up opportunities to peer into a multitude of biological processes at the cellular scale in their native microenvironments. Laser scanning microscopy (LSM) coupled with targeted fluorescent proteins has become an indispensable tool to enable dynamic imaging in vivo at high temporal and spatial resolutions. In the last few decades, the technique has been translated from imaging cells in thin samples to mapping cells in the thick biological tissue of living organisms. Here, we sought to provide a concise overview of the design considerations of a LSM that enables cellular and subcellular imaging in deep tissue. Individual components under review include: long working distance microscope objectives, laser scanning technologies, adaptive optics devices, beam shaping technologies and photon detectors, with an emphasis on more recent advances. The review will conclude with the latest innovations in automated optical microscopy, which would impact tracking and quantification of heterogeneous populations of cells in vivo.

  16. Nanosecond laser coloration on stainless steel surface.

    PubMed

    Lu, Yan; Shi, Xinying; Huang, Zhongjia; Li, Taohai; Zhang, Meng; Czajkowski, Jakub; Fabritius, Tapio; Huttula, Marko; Cao, Wei

    2017-08-02

    In this work, we present laser coloration on 304 stainless steel using nanosecond laser. Surface modifications are tuned by adjusting laser parameters of scanning speed, repetition rate, and pulse width. A comprehensive study of the physical mechanism leading to the appearance is presented. Microscopic patterns are measured and employed as input to simulate light-matter interferences, while chemical states and crystal structures of composites to figure out intrinsic colors. Quantitative analysis clarifies the final colors and RGB values are the combinations of structural colors and intrinsic colors from the oxidized pigments, with the latter dominating. Therefore, the engineering and scientific insights of nanosecond laser coloration highlight large-scale utilization of the present route for colorful and resistant steels.

  17. A landmark-based 3D calibration strategy for SPM

    NASA Astrophysics Data System (ADS)

    Ritter, Martin; Dziomba, Thorsten; Kranzmann, Axel; Koenders, Ludger

    2007-02-01

    We present a new method for the complete three-dimensional (3D) calibration of scanning probe microscopes (SPM) and other high-resolution microscopes, e.g., scanning electron microscopes (SEM) and confocal laser scanning microscopes (CLSM), by applying a 3D micrometre-sized reference structure with the shape of a cascade slope-step pyramid. The 3D reference structure was produced by focused ion beam induced metal deposition. In contrast to pitch featured calibration procedures that require separate lateral and vertical reference standards such as gratings and step height structures, the new method includes the use of landmarks, which are well established in calibration and measurement tasks on a larger scale. However, the landmarks applied to the new 3D reference structures are of sub-micrometre size, the so-called 'nanomarkers'. The nanomarker coordinates are used for a geometrical calibration of the scanning process of SPM as well as of other instrument types such as SEM and CLSM. For that purpose, a parameter estimation routine involving three scale factors and three coupling factors has been developed that allows lateral and vertical calibration in only one sampling step. With this new calibration strategy, we are able to detect deviations of SPM lateral scaling errors as well as coupling effects causing, e.g., a lateral coordinate shift depending on the measured height position of the probe.

  18. A STED-FLIM microscope applied to imaging the natural killer cell immune synapse

    NASA Astrophysics Data System (ADS)

    Lenz, M. O.; Brown, A. C. N.; Auksorius, E.; Davis, D. M.; Dunsby, C.; Neil, M. A. A.; French, P. M. W.

    2011-03-01

    We present a stimulated emission depletion (STED) fluorescence lifetime imaging (FLIM) microscope, excited by a microstructured optical fibre supercontinuum source that is pumped by a femtosecond Ti:Sapphire-laser, which is also used for depletion. Implemented using a piezo-scanning stage on a laser scanning confocal fluorescence microscope system with FLIM realised using time correlated single photon counting (TCSPC), this provides convenient switching between confocal and STED-FLIM with spatial resolution down to below 60 nm. We will present our design considerations to make a robust instrument for biological applications including a comparison between fixed phase plate and spatial light modulator (SLM) approaches to shape the STED beam and the correlation of STED and confocal FLIM microscopy. Following our previous application of FLIM-FRET to study intercellular signalling at the immunological synapse (IS), we are employing STED microscopy to characterize the spatial distribution of cellular molecules with subdiffraction resolution at the IS. In particular, we are imaging cytoskeletal structure at the Natural Killer cell activated immune synapse. We will also present our progress towards multilabel STED microscopy to determine how relative spatial molecular organization, previously undetectable by conventional microscopy techniques, is important for NK cell cytotoxic function. Keywords: STED, Stimulated Emission Depletion Microscopy, Natural Killer (NK) cell, Fluorescence lifetime imaging, FLIM, Super resolution microscopy.

  19. Visualization of femtosecond laser pulse-induced microincisions inside crystalline lens tissue.

    PubMed

    Stachs, Oliver; Schumacher, Silvia; Hovakimyan, Marine; Fromm, Michael; Heisterkamp, Alexander; Lubatschowski, Holger; Guthoff, Rudolf

    2009-11-01

    To evaluate a new method for visualizing femtosecond laser pulse-induced microincisions inside crystalline lens tissue. Laser Zentrum Hannover e.V., Hannover, Germany. Lenses removed from porcine eyes were modified ex vivo by femtosecond laser pulses (wavelength 1040 nm, pulse duration 306 femtoseconds, pulse energy 1.0 to 2.5 microJ, repetition rate 100 kHz) to create defined planes at which lens fibers separate. The femtosecond laser pulses were delivered by a 3-dimension (3-D) scanning unit and transmitted by focusing optics (numerical aperture 0.18) into the lens tissue. Lens fiber orientation and femtosecond laser-induced microincisions were examined using a confocal laser scanning microscope (CLSM) based on a Rostock Cornea Module attached to a Heidelberg Retina Tomograph II. Optical sections were analyzed in 3-D using Amira software (version 4.1.1). Normal lens fibers showed a parallel pattern with diameters between 3 microm and 9 microm, depending on scanning location. Microincision visualization showed different cutting effects depending on pulse energy of the femtosecond laser. The effects ranged from altered tissue-scattering properties with all fibers intact to definite fiber separation by a wide gap. Pulse energies that were too high or overlapped too tightly produced an incomplete cutting plane due to extensive microbubble generation. The 3-D CLSM method permitted visualization and analysis of femtosecond laser pulse-induced microincisions inside crystalline lens tissue. Thus, 3-D CLSM may help optimize femtosecond laser-based procedures in the treatment of presbyopia.

  20. Recycling stainless steel orthodontic brackets with Er:YAG laser - An environmental scanning electron microscope and shear bond strength study.

    PubMed

    Chacko, Prince K; Kodoth, Jithesh; John, Jacob; Kumar, Kishore

    2013-07-01

    TO DETERMINE THE EFFICIENCY OF ERBIUM: Yttrium aluminum garnet (Er:YAG) laser with Environmental Scanning Electron Microscope (ESEM) and shear bond strength analysis as a method of recycling stainless steel orthodontic brackets and compare with other methods of recycling. Eighty samples of extracted premolar teeth bonded to SS brackets were tested for rebonded shear bond strength after recycling by four methods and compared with a control group of 20 samples. These 80 samples were randomized into four groups which were recycled by four methods, namely, sandblasting, thermal method, adhesive grinding by tungsten carbide bur, and Er: YAG laser method. After recycling, ESEM and shear bond strength analysis were used to analyze the efficiency of the recycling methods. ER: YAG laser group was found to be having the greatest bond strength among the recycled brackets (8.33±2.51 followed by the sandblasting at 6.12±1.12 MPa, thermal and electropolishing at 4.44±0.95 MPa, and lastly the adhesive grinding method at 3.08±1.07 MPa. The shear bond strength of Er: YAG laser group was found to be having no statistically significant difference with that of the control group (P>0.05 and had statistical signifance with sandblasting, thermal and electropolishing and adhesive grinding groups at P>0.001. ESEM analysis showed complete removal of adhesive from the brackets recycled with Er: YAG laser which mimicked that of the control group. ER: YAG laser (2940 nm) was found to be the most efficient method for recycling, followed by the sandblasting, thermal, and the tungsten carbide methods, which had the least shear bond strength value and is not fit for clinical usage.

  1. Use of a white light supercontinuum laser for confocal interference-reflection microscopy

    PubMed Central

    Chiu, L-D; Su, L; Reichelt, S; Amos, WB

    2012-01-01

    Shortly after its development, the white light supercontinuum laser was applied to confocal scanning microscopy as a more versatile substitute for the multiple monochromatic lasers normally used for the excitation of fluorescence. This light source is now available coupled to commercial confocal fluorescence microscopes. We have evaluated a supercontinuum laser as a source for a different purpose: confocal interferometric imaging of living cells and artificial models by interference reflection. We used light in the range 460–700 nm where this source provides a reasonably flat spectrum, and obtained images free from fringe artefacts caused by the longer coherence length of conventional lasers. We have also obtained images of cytoskeletal detail that is difficult to see with a monochromatic laser. PMID:22432542

  2. Femtosecond laser-induced phase transformations in amorphous Cu77Ni6Sn10P7 alloy

    NASA Astrophysics Data System (ADS)

    Zhang, Y.; Liu, L.; Zou, G.; Chen, N.; Wu, A.; Bai, H.; Zhou, Y.

    2015-01-01

    In this study, the femtosecond laser-induced crystallization of CuNiSnP amorphous ribbons was investigated by utilizing an amplified Ti:sapphire laser system. X-ray diffraction and scanning electronic microscope were applied to examine the phase and morphology changes of the amorphous ribbons. Micromachining without crystallization, surface patterning, and selective crystallization were successfully achieved by changing laser parameters. Obvious crystallization occurred under the condition that the laser fluence was smaller than the ablation threshold, indicating that the structural evolution of the material depends strongly on the laser parameters. Back cooling method was used to inhibit heat accumulation; a reversible transformation between the disordered amorphous and crystalline phases can be achieved by using this method.

  3. A combined confocal and magnetic resonance microscope for biological studies

    NASA Astrophysics Data System (ADS)

    Majors, Paul D.; Minard, Kevin R.; Ackerman, Eric J.; Holtom, Gary R.; Hopkins, Derek F.; Parkinson, Christopher I.; Weber, Thomas J.; Wind, Robert A.

    2002-12-01

    Complementary data acquired with different microscopy techniques provide a basis for establishing a more comprehensive understanding of cell function in health and disease, particularly when results acquired with different methodologies can be correlated in time and space. In this article, a novel microscope is described for studying live cells simultaneously with both confocal scanning laser fluorescence optical microscopy and magnetic resonance microscopy. The various design considerations necessary for integrating these two complementary techniques are discussed, the layout and specifications of the instrument are given, and examples of confocal and magnetic resonance images of large frog cells and model tumor spheroids obtained with the compound microscope are presented.

  4. Photodynamic therapy and knocking out of single tumor cells by multiphoton excitation processes

    NASA Astrophysics Data System (ADS)

    Riemann, Iris; Fischer, Peter; Koenig, Karsten

    2004-09-01

    Near infrared (NIR) ultrashort laser pulses of 780 nm have been used to induce intracellular photodynamic reactions by nonlinear excitation of porphyrin photosensitizers. Intracellular accumulation and photobleaching of the fluorescent photosensitizers protoporphyrin IX and Photofrin (PF) have been studied by non-resonant two-photon fluorescence excitation of PF and aminolevulinic acid (ALA)-labeled Chinese hamster ovary (CHO) cells. To testify the efficacy of both substrates to induce irreversible destructive effects, the cloning efficiency (CE) of cells exposed to femtosecond pulses of a multiphoton laser scanning microscope (40x/1.3) was determined. In the case of Photofrin accumulation, CEs of 50% and 0% were obtained after 17 laserscans (2 mW?, 16 s/ frame) and 50 scans, respectively. All cells exposed to 50 scans died within 48h after laser exposure. 100 scans were required to induce lethal effects in ALA labeled cells. Sensitizer-free control cells could be scanned 250 times (1.1 h) and more without impact on the reproduction behavior, morphology, and vitality. In addition to the slow phototoxic effect by photooxidation processes, another destructive but immediate effect based on optical breakdown was induced when employing high intense NIR femtosecond laser beams. This was used to optically knock out single tumor cells in living mice (solid Ehrlich-Carcinoma) in a depth of 10 to 100 μm.

  5. Multimodal backside imaging of a microcontroller using confocal laser scanning and optical-beam-induced current imaging

    NASA Astrophysics Data System (ADS)

    Finkeldey, Markus; Göring, Lena; Schellenberg, Falk; Brenner, Carsten; Gerhardt, Nils C.; Hofmann, Martin

    2017-02-01

    Microscopy imaging with a single technology is usually restricted to a single contrast mechanism. Multimodal imaging is a promising technique to improve the structural information that could be obtained about a device under test (DUT). Due to the different contrast mechanisms of laser scanning microscopy (LSM), confocal laser scanning microscopy (CLSM) and optical beam induced current microscopy (OBICM), a combination could improve the detection of structures in integrated circuits (ICs) and helps to reveal their layout. While OBIC imaging is sensitive to the changes between differently doped areas and to semiconductor-metal transitions, CLSM imaging is mostly sensitive to changes in absorption and reflection. In this work we present the implementation of OBIC imaging into a CLSM. We show first results using industry standard Atmel microcontrollers (MCUs) with a feature size of about 250nm as DUTs. Analyzing these types of microcontrollers helps to improve in the field of side-channel attacks to find hardware Trojans, possible spots for laser fault attacks and for reverse engineering. For the experimental results the DUT is placed on a custom circuit board that allows us to measure the current while imaging it in our in-house built stage scanning microscope using a near infrared (NIR) laser diode as light source. The DUT is thinned and polished, allowing backside imaging through the Si-substrate. We demonstrate the possibilities using this optical setup by evaluating OBIC, LSM and CLSM images above and below the threshold of the laser source.

  6. The HVAC Challenges of Upgrading an Old Lab for High-end Light Microscopes

    PubMed Central

    Richard, R.; Martone, P.; Callahan, L.M.

    2014-01-01

    The University of Rochester Medical Center forms the centerpiece of the University of Rochester's health research, teaching, patient care, and community outreach missions. Within this large facility of over 5 million square feet, demolition and remodeling of existing spaces is a constant activity. With more than $145 million in federal research funding, lab space is frequently repurposed and renovated to support this work. The URMC Medical Center Facilities Organization supporting small to medium space renovations is constantly challenged and constrained by the existing mechanical infrastructure and budgets to deliver a renovated space that functions within the equipment environmental parameters. One recent project, sponsored by the URMC Shared Resources Laboratory, demonstrates these points. The URMC Light Microscopy Shared Resource Laboratory requested renovation of a 121 sq. ft. room in a 40 year old building which would enable placement of a laser capture microdissection microscope and a Pascal 5 laser scanning confocal microscope with the instruments separated by a blackout curtain. This poster discusses the engineering approach implemented to bring an older lab into the environmental specifications needed for the proper operation of the high-end light microscopes.

  7. DEVELOPMENT OF LiCo0.90Mg0.05Al0.05O2 THIN FILMS BY PULSED LASER DEPOSITION TECHNIQUE

    NASA Astrophysics Data System (ADS)

    Vasanthi, R.; Ruthmangani, I.; Manoravi, P.; Joseph, M.; Kesavamoorthy, R.; Sundar, C.; Selladurai, S.

    LiCo0.90Mg0.05Al0.05O2 bulk powders are synthesized using combustion process and made into a thin film by depositing on silicon wafer using a pulsed laser ablation technique. A comparative study by SEM (Scanning Electron Microscope) XRD (X-ray diffraction), Infrared spectroscopy and Raman Spectroscopy is performed on both bulk and PLD thin films.

  8. The development of laser surgery and medicine in China

    NASA Astrophysics Data System (ADS)

    Chen, Mingzhe

    2005-07-01

    The first Chinese ruby laser was created in 1961 and it was adopted for the retina coagulation experiment in 1965. Since 1970's, lasers had been widely applied clinically including the diseases suitable to physical therapy or acupuncture. The Chinese HpD was first produced in 1981 and first case of PDT was treated using Chinese HpD and Chinese lasers in the same year. Its success brought attention establishing a research group supported by the government in 1982. A nationwide systemic research project on PDT was then carried out. The step taken for PDT also accelerated the development of various fields of laser medicine and surgery. Laser treatments had been commonly adopted in the clinics and hospitals for the diseases of the superficial lesions and the lesions can be reached by the endoscopes non-invasively in 1980's. Since 1990's, the interventional laser therapies adopted mainly were percutaneous laser angioplasty, laser treatments through laparoscope, thoracoscope, arthroscope, neuro-endoscope etc. Ultrasound guided percutaneous laser heat coagulation for small hepatic cancer revealed good results and ultrasound guided percutaneous PDT for advanced large liver cancer revealed unexpected results after five years follow-up. At present: There are more long-term follow-up patients in the clinical trial; more advanced commercial available lasers and new techniques are adopted. Since the popularization of scanning electron microscope, laser scanning confocal microscope, laser induced auto-fluorescence system, high sensitivity fluorescence microscopic imaging system etc. in the laboratories, the basic studies can be more advanced and some times, the sub-cellular level can be reached; ultra-structure histo-morphology and gene studies are involved. In dermatology, Q-switched Alexandrite laser and other Q-switched lasers are used mainly for the treatment of skin pigmentation and vascular diseases; pulsed dye laser, ultra-pulsed CO2 laser are used in resurfacing, facial acne scar, osmidrosis etc. For ophthalmology, excimer laser are used for myopia or hyperopia; argon green laser, krypton yellow laser are adopted in coagulation for retinal detachment and neovascularization etc. Lasers are often used for the canaliculoplasty in the lacrimal canal. Low level lasers had been used very often on the acupuncture points and for many chronic diseases. Intravascular low level laser irradiation adopting semiconductor lasers and He-Ne laser were reported to use for comatose patients, schizophrenia, vascular dementia, Alzeimer"s disease and coronary disease. Reports from laboratory studies in the field of low level laser demonstrated the stimulation effect on the cells and immunology system; inhibitory effect on proliferation; it improved the biomedical data in hemorrheology; promoted the spinal motor nerve cell function, axonal regeneration; increased epidermal Langerhams cell to improve the antigen function, increased myocardial capillary permeability. Intra-coronary low power red laser irradiation assisted coronary interventional therapy showed its prevention effect on restenosis. Studies about the effects of various kinds of lasers, their wavelength, power densities and doses on various kinds of tissues were reported.

  9. EVALUATION OF CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: APPLICATIONS FOR IMAGING MORPHOLOGY AND DEATH IN EMBRYOS AND REPRODUCTIVE TISSUE/ORGANS

    EPA Science Inventory

    The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. It is remarkable that procedures to test the performance of these machines are not done routinely by most investigators and thus many of the machines in the field are working at level...

  10. Crystallization Behavior of Perovskite in the Synthesized High-Titanium-Bearing Blast Furnace Slag Using Confocal Scanning Laser Microscope

    NASA Astrophysics Data System (ADS)

    Hu, Meilong; Liu, Lu; Lv, Xuewei; Bai, Chenguang; Zhang, Shengfu

    2014-01-01

    The isothermal phase composition of high-titanium-bearing slag (23 mass pct TiO2) under an argon atmosphere during cooling process from 1723 K (1450 °C) was calculated by FactSage.6.3 (CRCT-ThermFact Inc., Montréal, Canada). Three main phases, which were perovskite, titania spinel, and clinopyroxene, could form during the cooling process and they precipitated at 1713 K, 1603 K, and 1498 K (1440 °C, 1330 °C, and 1225 °C), respectively. The nonisothermal crystallization process of perovskite in synthesized high-titanium-bearing slag was studied in situ by a confocal scanning laser microscope (CSLM) with cooling rate of 30 K/min. The results showed that the primary phase was perovskite that precipitated at 1703 K (1430 °C). The whole precipitation and growth process of perovskite was obtained, whereas other phases formed as glass under the current experimental conditions. Perovskite grew along a specific growth track and finally appeared with snowflake morphology. The growing kinetics of perovskite formation from molten slag were also mentioned.

  11. Histochemical study of trans-polyisoprene accumulation by spectral confocal laser scanning microscopy and a specific dye showing fluorescence solvatochromism in the rubber-producing plant, Eucommia ulmoides Oliver.

    PubMed

    Nakazawa, Yoshihisa; Takeda, Tsuyoshi; Suzuki, Nobuaki; Hayashi, Tatsushi; Harada, Yoko; Bamba, Takeshi; Kobayashi, Akio

    2013-09-01

    A microscopic technique combining spectral confocal laser scanning microscopy with a lipophilic fluorescent dye, Nile red, which can emit trans-polyisoprene specific fluorescence, was developed, and unmixed images of synthesized trans-polyisoprene in situ in Eucommia ulmoides were successfully obtained. The images showed that trans-polyisoprene was initially synthesized as granules in non-articulated laticifers that changed shape to fibers during laticifer maturation. Non-articulated laticifers are developed from single laticiferous cells, which are differentiated from surrounding parenchyma cells in the cambium. Therefore, these observations suggested that trans-polyisoprene biosynthesis first started in laticifer cells as granules and then the granules accumulated and fused in the inner space of the laticifers over time. Finally, laticifers were filled with the synthesized trans-polyisoprene, which formed a fibrous structure fitting the laticifers shape. Both trans- and cis-polyisoprene are among the most important polymers naturally produced by plants, and this microscopic technique combined with histological study should provide useful information in the fields of plant histology, bioindustry and phytochemistry.

  12. Remineralization Potential of Three Different Dentifrices using Raman Spectroscopy and Confocal Laser Scanning Microscope.

    PubMed

    Job, Tisson V; Narayana, Girish T; Venkappa, Kishan K; Nathan, K Binu; Ahsan, Shameem; Harikaran, Jayakkodi

    2018-04-01

    Aim: The aim of this study was to compare the remineralization potential of three different dentifrices using Raman spectroscopy and confocal laser scanning microscopy (CLSM). Materials and methods: Totally, 30 extracted intact impacted third molar teeth were selected and the crown of each tooth in a group was separated from the root and longitudinally sectioned into four parts with each section under a subgroup, of which one section was an untreated section, the second and the third sections were demineralized in a demineralizing solution, and the third section was remineralized after demineralization. The teeth in the three groups were demineralized for 4 days and then treated with 0.21% sodium fluoride dentifrice with trical-cium phosphate, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), and NovaMin for 14 days, following which the teeth surfaces were studied using Raman spec-troscopy and CLSM to assess the remineralization potential of the three dentifrices. The data were recorded and analyzed statistically. Results: Raman spectroscopic analysis revealed better remin-eralization with CPP-ACP, which was statistically significant from the groups treated with the NovaMin dentifrice and the fluoride-containing dentifrice.Confocal laser scanning microscopic examination also revealed significant differences between the three groups with the NovaMin-containing dentifrice demonstrating a greater remineralization of the surface when compared with the CPP-ACP dentifrice. The teeth samples treated with fluoride-containing dentifrice demonstrated the least reminer-alization among the three groups. Conclusion: It can be concluded that the demineralized samples of teeth treated with CPP-ACP showed the highest concentration of phosphate ions when analyzed using Raman spectroscopy, and the microscopic examination using confocal laser revealed a better surface remineralization of the demin-eralized samples when treated with the NovaMin technology. Clinical significance: There is a great need to find ways to enhance the remineralization process and transfer such knowledge into clinical therapy to alter caries balance for the better, especially in individuals with a high cariogenic bacterial challenge. Keywords: Casein phosphopeptide-amorphous calcium phosphate, Fluoride, NovaMin, Remineralization, Tricalcium phosphate.

  13. Evolution of Inclusions During the 1473 K (1200 °C) Heating Process of EH36 Shipbuilding Steel

    NASA Astrophysics Data System (ADS)

    Wang, Qiyu; Zou, Xiaodong; Matsuura, Hiroyuki; Wang, Cong

    2018-02-01

    Evolution behaviors of inclusions of EH36 shipbuilding steel during 1473 K (1200 °C) heating have been studied in conjunction with ex situ scanning electron microscope (SEM) examination and in situ confocal scanning laser microscopy (CSLM) observations. It has been found that Al-Ca-O-S complex inclusions dominate the particles in the cast billet. However, TiN inclusions are profusely populated after heating. Moreover, possible strategies governing austenite growth are offered here.

  14. Microstructure and physical properties of laser Zn modified amorphous-nanocrystalline coating on a titanium alloy

    NASA Astrophysics Data System (ADS)

    Li, Jia-Ning; Gong, Shui-Li; Shi, Yi-Ning; Suo, Hong-Bo; Wang, Xi-Chang; Deng, Yun-Hua; Shan, Fei-Hu; Li, Jian-Quan

    2014-02-01

    A Zn modified amorphous-nanocrystalline coating was fabricated on a Ti-6Al-4V alloy by laser cladding of the Co-Ti-B4C-Zn-Y2O3 mixed powders. Such coating was researched by means of a scanning electron microscope (SEM) and a high resolution transmission electron microscope (HRTEM), etc. Experimental results indicated that the Co5Zn21 and TiB2 nanocrystalline phases were produced through in situ metallurgical reactions, which blocked the motion of dislocation, and TiB2 grew along (010), (111) and (024). The Co5Zn21 nanocrystals were produced attached to the ceramics, which mainly consisted of the Co nanoparticles embedded in a heterogeneous zinc, and had varied crystalline orientations.

  15. [INVITED] Laser treatment of Inconel 718 alloy and surface characteristics

    NASA Astrophysics Data System (ADS)

    Yilbas, B. S.; Ali, H.; Al-Aqeeli, N.; Karatas, C.

    2016-04-01

    Laser surface texturing of Inconel 718 alloy is carried out under the high pressure nitrogen assisting gas. The combination of evaporation and melting at the irradiated surface is achieved by controlling the laser scanning speed and the laser output power. Morphological and metallurgical changes in the treated surface are analyzed using the analytical tools including optical, electron scanning, and atomic force microscopes, energy dispersive spectroscopy, and X-ray diffraction. Microhardnes and friction coefficient of the laser treated surface are measured. Residual stress formed in the surface region is determined from the X-ray diffraction data. Surface hydrophobicity of the laser treated layer is assessed incorporating the sessile drop method. It is found that laser treated surface is free from large size asperities including cracks and the voids. Surface microhardness increases significantly after the laser treatment process, which is attributed to the dense layer formation at the surface under the high cooling rates, dissolution of Laves phase in the surface region, and formation of nitride species at the surface. Residual stress formed is compressive in the laser treated surface and friction coefficient reduces at the surface after the laser treatment process. The combination of evaporation and melting at the irradiated surface results in surface texture composes of micro/nano-poles and pillars, which enhance the surface hydrophobicity.

  16. Surface characterization of carbon fiber polymer composites and aluminum alloys after laser interference structuring

    DOE PAGES

    Sabau, Adrian S.; Greer, Clayton M.; Chen, Jian; ...

    2016-05-03

    Here, the increasing use of Carbon Fiber-reinforced Polymer matrix Composites (CFPC) and aluminum alloys as lightweight materials in the automotive and aerospace industries demands enhanced surface preparation and control of surface morphology prior to joining. In this study, surfaces of both composite and aluminum were prepared for joining using an Nd:YAG laser in a two-beam interference setup, enabling the (a) structuring of the AL 5182 surface, (b) removal of the resin layer on top of carbon fibers, and (c) structuring of the carbon fibers. CFPC specimens of T700S carbon fiber, Prepreg - T8 3 epoxy, 5 ply thick, 0/90o plaquesmore » were used. The effect of laser fluence, scanning speed, and number of shots-per-spot was investigated on the removal rate of the resin without an excessive damage of the fibers. Optical micrographs, 3D imaging, and scanning electron microscope (SEM) imaging were used to study the effect of the laser processing on surface morphology.« less

  17. Miniature near-infrared dual-axes confocal microscope utilizing a two-dimensional microelectromechanical systems scanner

    PubMed Central

    Liu, Jonathan T. C.; Mandella, Michael J.; Ra, Hyejun; Wong, Larry K.; Solgaard, Olav; Kino, Gordon S.; Piyawattanametha, Wibool; Contag, Christopher H.; Wang, Thomas D.

    2007-01-01

    The first, to our knowledge, miniature dual-axes confocal microscope has been developed, with an outer diameter of 10 mm, for subsurface imaging of biological tissues with 5–7 μm resolution. Depth-resolved en face images are obtained at 30 frames per second, with a field of view of 800 × 100 μm, by employing a two-dimensional scanning microelectromechanical systems mirror. Reflectance and fluorescence images are obtained with a laser source at 785 nm, demonstrating the ability to perform real-time optical biopsy. PMID:17215937

  18. Characterization of a Hybrid Optical Microscopy/Laser Ablation Liquid Vortex Capture/Electrospray Ionization System for Mass Spectrometry Imaging

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

    Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.

    Herein, a commercial optical microscope, laser microdissection instrument was coupled with an electrospray ionization mass spectrometer via a low profile liquid vortex capture probe to yield a hybrid optical microscopy/mass spectrometry imaging system. The instrument has bright-field and fluorescence microscopy capabilities in addition to a highly focused UV laser beam that is utilized for laser ablation of samples. With this system, material laser ablated from a sample using the microscope was caught by a liquid vortex capture probe and transported in solution for analysis by electrospray ionization mass spectrometry. Both lane scanning and spot sampling mass spectral imaging modes weremore » used. The smallest area the system was able to ablate was ~0.544 μm × ~0.544 μm, achieved by oversampling of the smallest laser ablation spot size that could be obtained (~1.9 μm). With use of a model photoresist surface, known features as small as ~1.5 μm were resolved. The capabilities of the system with real world samples were demonstrated first with a blended polymer thin film containing poly(2-vinylpyridine) and poly(N-vinylcarbazole). Using spot sampling imaging, sub-micrometer sized features (0.62, 0.86, and 0.98 μm) visible by optical microscopy were clearly distinguished in the mass spectral images. A second real world example showed the imaging of trace amounts of cocaine in mouse brain thin tissue sections. Lastly, with use of a lane scanning mode with ~6 μm × ~6 μm data pixels, features in the tissue as small as 15 μm in size could be distinguished in both the mass spectral and optical images.« less

  19. Characterization of a Hybrid Optical Microscopy/Laser Ablation Liquid Vortex Capture/Electrospray Ionization System for Mass Spectrometry Imaging

    DOE PAGES

    Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.

    2015-10-22

    Herein, a commercial optical microscope, laser microdissection instrument was coupled with an electrospray ionization mass spectrometer via a low profile liquid vortex capture probe to yield a hybrid optical microscopy/mass spectrometry imaging system. The instrument has bright-field and fluorescence microscopy capabilities in addition to a highly focused UV laser beam that is utilized for laser ablation of samples. With this system, material laser ablated from a sample using the microscope was caught by a liquid vortex capture probe and transported in solution for analysis by electrospray ionization mass spectrometry. Both lane scanning and spot sampling mass spectral imaging modes weremore » used. The smallest area the system was able to ablate was ~0.544 μm × ~0.544 μm, achieved by oversampling of the smallest laser ablation spot size that could be obtained (~1.9 μm). With use of a model photoresist surface, known features as small as ~1.5 μm were resolved. The capabilities of the system with real world samples were demonstrated first with a blended polymer thin film containing poly(2-vinylpyridine) and poly(N-vinylcarbazole). Using spot sampling imaging, sub-micrometer sized features (0.62, 0.86, and 0.98 μm) visible by optical microscopy were clearly distinguished in the mass spectral images. A second real world example showed the imaging of trace amounts of cocaine in mouse brain thin tissue sections. Lastly, with use of a lane scanning mode with ~6 μm × ~6 μm data pixels, features in the tissue as small as 15 μm in size could be distinguished in both the mass spectral and optical images.« less

  20. Nd:YOV4 laser polishing on WC-Co HVOF coating

    NASA Astrophysics Data System (ADS)

    Giorleo, L.; Ceretti, E.; Montesano, L.; La Vecchia, G. M.

    2017-10-01

    WC/Co coatings are widely applied to different types of components due to their extraordinary performance properties including high hardness and wear properties. In industrial applications High Velocity Oxy-Fuel (HVOF) technique is extensively used to deposit hard metal coatings. The main advantage of HVOF compared to other thermal spray techniques is the ability to accelerate the melted powder particles of the feedstock material at a relatively high velocity, leading to obtain good adhesion and low porosity level. However, despite the mentioned benefits, the surface finish quality of WC-Co HVOF coatings results to be poor (Ra higher than 5 µm) thus a mechanical polishing process is often needed. The main problem is that the high hardness of coating leads the polishing process expensive in terms of time and tool wear; moreover polishing becomes difficult and not always possible in case of limited accessibility of a part, micro dimensions or undercuts. Nowadays a different technique available to improve surface roughness is the laser polishing process. The polishing principle is based on focused radiation of a laser beam that melts a microscopic layer of surface material. Compared to conventional polishing process (as grinding) it ensures the possibility of avoiding tool wear, less pollution (no abrasive or liquids), no debris, less machining time and coupled with a galvo system it results to be more suitable in case of 3D complex workpieces. In this paper laser polishing process executed with a Nd:YOV4 Laser was investigated: the effect of different process parameters as initial coating morphology, laser scan speed and loop cycles were tested. Results were compared by a statistical approach in terms of average roughness along with a morphological analysis carried out by Scanning Electron Microscope (SEM) investigation coupled with EDS spectra.

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

    Sabau, Adrian S.; Greer, Clayton M.; Chen, Jian

    Here, the increasing use of Carbon Fiber-reinforced Polymer matrix Composites (CFPC) and aluminum alloys as lightweight materials in the automotive and aerospace industries demands enhanced surface preparation and control of surface morphology prior to joining. In this study, surfaces of both composite and aluminum were prepared for joining using an Nd:YAG laser in a two-beam interference setup, enabling the (a) structuring of the AL 5182 surface, (b) removal of the resin layer on top of carbon fibers, and (c) structuring of the carbon fibers. CFPC specimens of T700S carbon fiber, Prepreg - T8 3 epoxy, 5 ply thick, 0/90o plaquesmore » were used. The effect of laser fluence, scanning speed, and number of shots-per-spot was investigated on the removal rate of the resin without an excessive damage of the fibers. Optical micrographs, 3D imaging, and scanning electron microscope (SEM) imaging were used to study the effect of the laser processing on surface morphology.« less

  2. Nd:YAG laser ablation and acid resistance of enamel.

    PubMed

    Kwon, Yong Hoon; Kwon, Oh-Won; Kim, Hyung-Il; Kim, Kyo-Han

    2003-09-01

    The acid resistance of Nd:YAG laser-ablated enamel surfaces was studied by evaluating crystal structure, mineral distribution, and fluorescence radiance and image in the present study. For comparison, 37% phosphoric acid etching was performed. The formation of beta-tricalcium phosphate (beta-TCP) was confirmed in the laser-ablated surface. The Ca/P ratio increased after ablation due to mineral re-distribution. In contrast, the Ca/P ratio decreased after acid etching due to mineral loss. The laser-ablated enamels showed a smaller increase of fluorescence radiances and less clear laser confocal scanning microscope images than those observed in the acid-etched enamels. The former suggests a minimized mineral loss. The Nd:YAG laser irradiation will enhance the acid resistance and retard the carious progression in enamel.

  3. Eyecup scope—optical recordings of light stimulus-evoked fluorescence signals in the retina

    PubMed Central

    Hausselt, Susanne E.; Breuninger, Tobias; Castell, Xavier; Denk, Winfried; Margolis, David J.; Detwiler, Peter B.

    2009-01-01

    Dendritic signals play an essential role in processing visual information in the retina. To study them in neurites too small for electrical recording, we developed an instrument that combines a multi-photon (MP) microscope with a through-the-objective high-resolution visual stimulator. An upright microscope was designed that uses the objective lens for both MP imaging and delivery of visual stimuli to functionally intact retinal explants or eyecup preparations. The stimulator consists of a miniature liquid-crystal-on-silicon display coupled into the optical path of an infrared-excitation laser-scanning microscope. A pair of custom-made dichroic filters allows light from the excitation laser and three spectral bands (‘colors’) from the stimulator to reach the retina, leaving two intermediate bands for fluorescence imaging. Special optics allow displacement of the stimulator focus relative to the imaging focus. Spatially resolved changes in calcium-indicator fluorescence in response to visual stimuli were recorded in dendrites of different types of mammalian retinal neurons. PMID:19023590

  4. The role of biomineralization in microbiologically influenced corrosion

    NASA Technical Reports Server (NTRS)

    Little, B.; Wagner, P.; Hart, K.; Ray, R.; Lavoie, D.; Nealson, K.; Aguilar, C.

    1998-01-01

    Synthetic iron oxides (goethite, alpha-FeO.OH; hematite, Fe2O3; and ferrihydrite, Fe(OH)3) were used as model compounds to simulate the mineralogy of surface films on carbon steel. Dissolution of these oxides exposed to pure cultures of the metal-reducing bacterium, Shewanella putrefaciens, was followed by direct atomic absorption spectroscopy measurement of ferrous iron coupled with microscopic analyses using confocal laser scanning and environmental scanning electron microscopies. During an 8-day exposure the organism colonized mineral surfaces and reduced solid ferric oxides to soluble ferrous ions. Elemental composition, as monitored by energy dispersive x-ray spectroscopy, indicated mineral replacement reactions with both ferrihydrite and goethite as iron reduction occurred. When carbon steel electrodes were exposed to S. putrefaciens, microbiologically influenced corrosion was demonstrated electrochemically and microscopically.

  5. Selective treatment of carious dentin using a mid-infrared tunable pulsed laser at 6 μm wavelength range

    NASA Astrophysics Data System (ADS)

    Saiki, Masayuki; Ishii, Katsunori; Yoshikawa, Kazushi; Yasuo, Kenzo; Yamamoto, Kazuyo; Awazu, Kunio

    2011-03-01

    Optical technologies have good potential for caries detection, prevention, excavation, and the realization of minimal intervention dentistry. This study aimed to develop a selective excavation technique of carious tissue using the specific absorption in 6 μm wavelength range. Bovine dentin demineralized with lactic acid solution was used as a carious dentin model. A mid-infrared tunable pulsed laser was obtained by difference-frequency generation technique. The wavelength was tuned to 6.02 and 6.42 μm which correspond to absorption bands called amide I and amide II, respectively. The laser delivers 5 ns pulse width at a repetition rate of 10 Hz. The morphological change after irradiation was observed with a scanning electron microscope, and the measurement of ablation depth was performed with a confocal laser microscope. At λ = 6.02 μm and the average power density of 15 W/cm2, demineralized dentin was removed selectively with less-invasive effect on sound dentin. The wavelength of 6.42 μm also showed the possibility of selective removal. High ablation efficiency and low thermal side effect were observed using the nanosecond pulsed laser with λ = 6.02 μm. In the near future, development of compact laser device will open the minimal invasive laser treatment to the dental clinic.

  6. Study and development of 22 kW peak power fiber coupled short pulse Nd:YAG laser for cleaning applications

    NASA Astrophysics Data System (ADS)

    Choubey, Ambar; Vishwakarma, S. C.; Vachhani, D. M.; Singh, Ravindra; Misra, Pushkar; Jain, R. K.; Arya, R.; Upadhyaya, B. N.; Oak, S. M.

    2014-11-01

    Free running short pulse Nd:YAG laser of microsecond pulse duration and high peak power has a unique capability to ablate material from the surface without heat propagation into the bulk. Applications of short pulse Nd:YAG lasers include cleaning and restoration of marble, stones, and a variety of metals for conservation. A study on the development of high peak power short pulses from Nd:YAG laser along with its cleaning and conservation applications has been performed. A pulse energy of 1.25 J with 55 μs pulse duration and a maximum peak power of 22 kW has been achieved. Laser beam has an M2 value of ~28 and a pulse-to-pulse stability of ±2.5%. A lower value of M2 means a better beam quality of the laser in multimode operation. A top hat spatial profile of the laser beam was achieved at the exit end of 200 μm core diameter optical fiber, which is desirable for uniform cleaning. This laser system has been evaluated for efficient cleaning of surface contaminations on marble, zircaloy, and inconel materials for conservation with cleaning efficiency as high as 98%. Laser's cleaning quality and efficiency have been analysed by using a microscope, a scanning electron microscope (SEM), and X-ray photon spectroscopy (XPS) measurements.

  7. Two-photon excitation laser scanning microscopy of rabbit nasal septal cartilage following Nd:YAG-laser-mediated stress relaxation

    NASA Astrophysics Data System (ADS)

    Kim, Charlton C.; Wallace, Vincent P.; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-04-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within rabbit nasal septal cartilage tissue over a 12-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation. During laser irradiation surface temperature, stress relaxation, and diffuse reflectance, were measured dynamically. Each specimen received one or two sequential laser exposures. The cartilage reached a peak surface temperature of about 61 degrees C during irradiation. Cartilage denatured in 50 percent EtOH was used as a positive control. TPM was performed to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns, immediately following laser exposure, and also following 12 days in culture. Few differences in the pattern or intensity of fluorescence was observed between controls and irradiated specimens imaged immediately following exposure, regardless of the number of laser pulses. However, following twelve days in tissue culture, the irradiated specimens increase, whereas the native tissue diminishes, in intensity and distribution of fluorescence in the cytoplasm. In contrast, the positive control shows only extracellular matrices and empty lacuna, feature consistent with cell membrane lysis.

  8. Silver Nanoscale Hexagonal Column Chips for Detecting Cell-free DNA and Circulating Nucleosomes in Cancer Patients.

    PubMed

    Ito, Hiroaki; Hasegawa, Katsuyuki; Hasegawa, Yuuki; Nishimaki, Tadashi; Hosomichi, Kazuyoshi; Kimura, Satoshi; Ohba, Motoi; Yao, Hiroshi; Onimaru, Manabu; Inoue, Ituro; Inoue, Haruhiro

    2015-05-21

    Blood tests, which are commonly used for cancer screening, generally have low sensitivity. Here, we developed a novel rapid and simple method to generate silver nanoscale hexagonal columns (NHCs) for use in surface-enhanced Raman scattering (SERS). We reported that the intensity of SERS spectra of clinical serum samples obtained from gastrointestinal cancer patients is was significantly higher than that of SERS spectra of clinical serum samples obtained from non-cancer patients. We estimated the combined constituents on silver NHCs by using a field emission-type scanning electron microscope, Raman microscopes, and a 3D laser scanning confocal microscope. We obtained the Raman scattering spectra of samples of physically fractured cells and clinical serum. No spectra were obtained for chemically lysed cultured cells and DNA, RNA, and protein extracted from cultured cells. We believe that our method, which uses SERS with silver NHCs to detect circulating nucleosomes bound by methylated cell-free DNA, may be successfully implemented in blood tests for cancer screening.

  9. Hybrid multiphoton volumetric functional imaging of large-scale bioengineered neuronal networks

    NASA Astrophysics Data System (ADS)

    Dana, Hod; Marom, Anat; Paluch, Shir; Dvorkin, Roman; Brosh, Inbar; Shoham, Shy

    2014-06-01

    Planar neural networks and interfaces serve as versatile in vitro models of central nervous system physiology, but adaptations of related methods to three dimensions (3D) have met with limited success. Here, we demonstrate for the first time volumetric functional imaging in a bioengineered neural tissue growing in a transparent hydrogel with cortical cellular and synaptic densities, by introducing complementary new developments in nonlinear microscopy and neural tissue engineering. Our system uses a novel hybrid multiphoton microscope design combining a 3D scanning-line temporal-focusing subsystem and a conventional laser-scanning multiphoton microscope to provide functional and structural volumetric imaging capabilities: dense microscopic 3D sampling at tens of volumes per second of structures with mm-scale dimensions containing a network of over 1,000 developing cells with complex spontaneous activity patterns. These developments open new opportunities for large-scale neuronal interfacing and for applications of 3D engineered networks ranging from basic neuroscience to the screening of neuroactive substances.

  10. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    NASA Astrophysics Data System (ADS)

    Darvin, M. E.; Richter, H.; Zhu, Y. J.; Meinke, M. C.; Knorr, F.; Gonchukov, S. A.; Koenig, K.; Lademann, J.

    2014-07-01

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted.

  11. Detection of endolithic spatial distribution in marble stone.

    PubMed

    Casanova Municchia, A; Percario, Z; Caneva, G

    2014-10-01

    The penetration of endolithic microorganisms, which develop to depths of several millimetres or even centimetres into the stone, and the diffusion of their extracellular substances speeds up the stone deterioration process. The aim of this study was to investigate, using a confocal laser scanning microscopy with a double-staining, a marble rock sample by observing the endolithic spatial distribution and quantifying the volume they occupied within the stone, in order to understand the real impact of these microorganisms on the conservation of stone monuments. Often the only factors taken into account by biodeterioration studies regarding endolithic microorganisms, are spread and depth of penetration. Despite the knowledge of three-dimensional spatial distribution and quantification of volume, it is indispensable to understand the real damage caused by endolithic microorganisms to stone monuments. In this work, we analyze a marble rock sample using a confocal laser scanning microscopy stained with propidium iodide and Concavalin-A conjugate with the fluorophore Alexa Fluor 488, comparing these results with other techniques (SEM microscope, microphotographs of polished cross-sections and thin-section, PAS staining methods), An image analysis approach has also been applied. The use of confocal laser scanning microscopy with double staining shows clear evidence of the presence of endolithic microorganisms (cyanobacteria and fungi) as well as the extracellular polymeric substance matrix in a three-dimensional architecture as part of the rock sample, this technique, therefore, seems very useful when applied to restoration interventions on stone monuments when endolithic growth is suspected. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

  12. Thermally-induced voltage alteration for analysis of microelectromechanical devices

    DOEpatents

    Walraven, Jeremy A.; Cole, Jr., Edward I.

    2002-01-01

    A thermally-induced voltage alteration (TIVA) apparatus and method are disclosed for analyzing a microelectromechanical (MEM) device with or without on-board integrated circuitry. One embodiment of the TIVA apparatus uses constant-current biasing of the MEM device while scanning a focused laser beam over electrically-active members therein to produce localized heating which alters the power demand of the MEM device and thereby changes the voltage of the constant-current source. This changing voltage of the constant-current source can be measured and used in combination with the position of the focused and scanned laser beam to generate an image of any short-circuit defects in the MEM device (e.g. due to stiction or fabrication defects). In another embodiment of the TIVA apparatus, an image can be generated directly from a thermoelectric potential produced by localized laser heating at the location of any short-circuit defects in the MEM device, without any need for supplying power to the MEM device. The TIVA apparatus can be formed, in part, from a scanning optical microscope, and has applications for qualification testing or failure analysis of MEM devices.

  13. Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy

    PubMed Central

    Fernández, A.; Grüner-Nielsen, L.; Andreana, M.; Stadler, M.; Kirchberger, S.; Sturtzel, C.; Distel, M.; Zhu, L.; Kautek, W.; Leitgeb, R.; Baltuska, A.; Jespersen, K.; Verhoef, A.

    2017-01-01

    A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy. PMID:28856032

  14. Super-Resolution Scanning Laser Microscopy Based on Virtually Structured Detection

    PubMed Central

    Zhi, Yanan; Wang, Benquan; Yao, Xincheng

    2016-01-01

    Light microscopy plays a key role in biological studies and medical diagnosis. The spatial resolution of conventional optical microscopes is limited to approximately half the wavelength of the illumination light as a result of the diffraction limit. Several approaches—including confocal microscopy, stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy, and structured illumination microscopy—have been established to achieve super-resolution imaging. However, none of these methods is suitable for the super-resolution ophthalmoscopy of retinal structures because of laser safety issues and inevitable eye movements. We recently experimentally validated virtually structured detection (VSD) as an alternative strategy to extend the diffraction limit. Without the complexity of structured illumination, VSD provides an easy, low-cost, and phase artifact–free strategy to achieve super-resolution in scanning laser microscopy. In this article we summarize the basic principles of the VSD method, review our demonstrated single-point and line-scan super-resolution systems, and discuss both technical challenges and the potential of VSD-based instrumentation for super-resolution ophthalmoscopy of the retina. PMID:27480461

  15. Optimizing pulse compressibility in completely all-fibered Ytterbium chirped pulse amplifiers for in vivo two photon laser scanning microscopy.

    PubMed

    Fernández, A; Grüner-Nielsen, L; Andreana, M; Stadler, M; Kirchberger, S; Sturtzel, C; Distel, M; Zhu, L; Kautek, W; Leitgeb, R; Baltuska, A; Jespersen, K; Verhoef, A

    2017-08-01

    A simple and completely all-fiber Yb chirped pulse amplifier that uses a dispersion matched fiber stretcher and a spliced-on hollow core photonic bandgap fiber compressor is applied in nonlinear optical microscopy. This stretching-compression approach improves compressibility and helps to maximize the fluorescence signal in two-photon laser scanning microscopy as compared with approaches that use standard single mode fibers as stretcher. We also show that in femtosecond all-fiber systems, compensation of higher order dispersion terms is relevant even for pulses with relatively narrow bandwidths for applications relying on nonlinear optical effects. The completely all-fiber system was applied to image green fluorescent beads, a stained lily-of-the-valley root and rat-tail tendon. We also demonstrated in vivo imaging in zebrafish larvae, where we simultaneously measure second harmonic and fluorescence from two-photon excited red-fluorescent protein. Since the pulses are compressed in a fiber, this source is especially suited for upgrading existing laser scanning (confocal) microscopes with multiphoton imaging capabilities in space restricted settings or for incorporation in endoscope-based microscopy.

  16. Textural Evolution During Micro Direct Metal Deposition of NiTi Alloy

    NASA Astrophysics Data System (ADS)

    Khademzadeh, Saeed; Bariani, Paolo F.; Bruschi, Stefania

    2018-03-01

    In this research, a micro direct metal deposition process, newly developed as a potential method for micro additive manufacturing was used to fabricate NiTi builds. The effect of scanning strategy on grain growth and textural evolution was investigated using scanning electron microscope equipped with electron backscattered diffraction detector. Investigations showed that, the angle between the successive single tracks has an important role in grain size distribution and textural evolution of NiTi phase. Unidirectional laser beam scanning pattern developed a fiber texture; conversely, a backward and forward scanning pattern developed a strong < {100} > ‖‖ RD texture on the surface of NiTi cubic samples produced by micro direct metal deposition.

  17. Textural Evolution During Micro Direct Metal Deposition of NiTi Alloy

    NASA Astrophysics Data System (ADS)

    Khademzadeh, Saeed; Bariani, Paolo F.; Bruschi, Stefania

    2018-07-01

    In this research, a micro direct metal deposition process, newly developed as a potential method for micro additive manufacturing was used to fabricate NiTi builds. The effect of scanning strategy on grain growth and textural evolution was investigated using scanning electron microscope equipped with electron backscattered diffraction detector. Investigations showed that, the angle between the successive single tracks has an important role in grain size distribution and textural evolution of NiTi phase. Unidirectional laser beam scanning pattern developed a fiber texture; conversely, a backward and forward scanning pattern developed a strong < {100} > ‖‖ RD texture on the surface of NiTi cubic samples produced by micro direct metal deposition.

  18. Development of a metrological atomic force microscope with a tip-tilting mechanism for 3D nanometrology

    NASA Astrophysics Data System (ADS)

    Kizu, Ryosuke; Misumi, Ichiko; Hirai, Akiko; Kinoshita, Kazuto; Gonda, Satoshi

    2018-07-01

    A metrological atomic force microscope with a tip-tilting mechanism (tilting-mAFM) has been developed to expand the capabilities of 3D nanometrology, particularly for high-resolution topography measurements at the surfaces of vertical sidewalls and for traceable measurements of nanodevice linewidth. In the tilting-mAFM, the probe tip is tilted from vertical to 16° at maximum such that the probe tip can touch and trace the vertical sidewall of a nanometer-scale structure; the probe of a conventional atomic force microscope cannot reach the vertical surface because of its finite cone angle. Probe displacement is monitored in three axes by using high-resolution laser interferometry, which is traceable to the SI unit of length. A central-symmetric 3D scanner with a parallel spring structure allows probe scanning with extremely low interaxial crosstalk. A unique technique for scanning vertical sidewalls was also developed and applied. The experimental results indicated high repeatability in the scanned profiles and sidewall angle measurements. Moreover, the 3D measurement of a line pattern was demonstrated, and the data from both sidewalls were successfully stitched together with subnanometer accuracy. Finally, the critical dimension of the line pattern was obtained.

  19. Quantitative and structural analyses of the in vitro and ex vivo biofilm-forming ability of dermatophytes.

    PubMed

    Brilhante, Raimunda Sâmia Nogueira; Correia, Edmilson Emanuel Monteiro; Guedes, Glaucia Morgana de Melo; Pereira, Vandbergue Santos; Oliveira, Jonathas Sales de; Bandeira, Silviane Praciano; Alencar, Lucas Pereira de; Andrade, Ana Raquel Colares de; Castelo-Branco, Débora de Souza Collares Maia; Cordeiro, Rossana de Aguiar; Pinheiro, Adriana de Queiroz; Chaves, Lúcio Jackson Queiroz; Pereira Neto, Waldemiro de Aquino; Sidrim, José Júlio Costa; Rocha, Marcos Fábio Gadelha

    2017-07-01

    The aim of this study was to evaluate the in vitro and ex vivo biofilm-forming ability of dermatophytes on a nail fragment. Initially, four isolates of Trichophyton rubrum, six of Trichophyton tonsurans, three of Trichophyton mentagrophytes, ten of Microsporum canis and three of Microsporum gypseum were tested for production biomass by crystal violet assay. Then, one strain per species presenting the best biofilm production was chosen for further studies by optical microscopy (Congo red staining), confocal laser scanning (LIVE/DEAD staining) and scanning electron (secondary electron) microscopy. Biomass quantification by crystal violet assay, optical microscope images of Congo red staining, confocal microscope and scanning electron microscope images revealed that all species studied are able to form biofilms both in vitro and ex vivo, with variable density and architecture. M. gypseum, T. rubrum and T. tonsurans produced robust biofilms, with abundant matrix and biomass, while M. canis produced the weakest biofilms compared to other species. This study sheds light on biofilms of different dermatophyte species, which will contribute to a better understanding of the pathophysiology of dermatophytosis. Further studies of this type are necessary to investigate the processes involved in the formation and composition of dermatophyte biofilms.

  20. One-step shaping of NiTi biomaterial by selective laser melting

    NASA Astrophysics Data System (ADS)

    Yang, Yongqiang; Huang, Yanlu; Wu, Wenhui

    2007-11-01

    NiTi alloy has excellent biocompatibility. This paper presents a novel technology of direct shaping of this promising biomaterial with selective laser melting (SLM). The frequently encountered defects of the SLM metal alloy parts such as non-fully melting, thermal deformation and balling were analyzed theoretically and experimentally, and the microstructure of the parts was analyzed on microscope. The results show that an appropriate selection of laser mode and scanning strategy assures a satisfying quality of the final parts; they also reveal that the SLM technology can be expected as a potential technology to directly manufacture the artificial implant of NiTi alloys.

  1. An experimental technique to repair cracked teeth using calcium phosphate, melted by a laser beam: an in vitro evaluation.

    PubMed

    Levy, G C; Koubi, G F

    1993-11-01

    Using a neodymium: yttrium-aluminum-garnet laser beam to seal vertical root fracture lines with tricalcium phosphate paste represents an alternative treatment for cracked teeth with noted clinical results. This article describes a study of the permeability of molten crystals of hydroxyapatite in the dentin of a cracked root after crack lines have been filled with a preparation of tricalcium phosphate melted by a neodymium: yttrium-aluminum-garnet laser beam. The morphology of the sealed cracks was analyzed under a scanning electron microscope that showed a deep fusion of tricalcium phosphate along crack lines.

  2. Physical and Microstructure Properties of MgAl2C2 Matrix Composite Coating on Titanium

    NASA Astrophysics Data System (ADS)

    Li, Peng

    2014-12-01

    This work is based on the dry sliding wear of the MgAl2C2-TiB2-FeSi composite coating deposited on a pure Ti using a laser cladding technique. Scanning electron microscope images indicate that the nanocrystals and amorphous phases are produced in such coating. X-ray diffraction result indicated that such coating mainly consists of MgAl2C2, Ti-B, Ti-Si, Fe-Al, Ti3SiC2, TiC and amorphous phases. The high resolution transmission electron microscope image indicated that the TiB nanorods were produced in the coating, which were surrounded by other fine precipitates, favoring the formation of a fine microstructure. With increase of the laser power from 0.85 kW to 1.00 kW, the micro-hardness decreased from 1350 1450 HV0.2 to 1200 1300 HV0.2. The wear volume loss of the laser clad coating was 1/7 of pure Ti.

  3. Differential polarization laser scanning microscopy: biological applications

    NASA Astrophysics Data System (ADS)

    Steinbach, G.; Besson, F.; Pomozi, I.; Garab, G.

    2005-09-01

    With the aid of a differential polarization (DP) apparatus, developed in our laboratory and attached to our laser scanning confocal microscope, we can measure the magnitude and spatial distribution of 8 different DP quantities: linear and circular dichroism (LD&CD), linear and circular anisotropy of the emission (R and CPL, confocal), fluorescence detected dichroisms (FDLD&FDCD, confocal), linear birefringence (LB), and the degree of polarization of fluorescence emission (P, confocal). The attachment uses high frequency modulation and subsequent demodulation, via lock-in amplifier, of the detected intensity values, and records and displays pixel-by-pixel the measured DP quantity. These microscopic DP data carry important physical information on the molecular architecture of anisotropically organized samples. Microscopic DP measurements are thought to be of particular importance in biology. In most biological samples anisotropy is difficult to determine with conventional, macroscopic DP measurements and microscopic variations are of special significance. In this paper, we describe the method of LB imaging. Using magnetically oriented isolated chloroplasts trapped in polyacrylamide gel, we demonstrate that LB can be determined with high sensitivity and good spatial resolution. Granal thylakoid membranes in edge-aligned orientation exhibited strong LB, with large variations in its sign and magnitude. In face-aligned position LB was considerably weaker, and tended to vanish when averaged for the whole image. The strong local variations are attributed to the inherent heterogeneity of the membranes, i.e. to their internal differentiation into multilamellar, stacked membranes (grana), and single thylakoids (stroma membranes). Further details and applications of our DP-LSM will be published elsewhere.

  4. Neodymium:yttrium-aluminum-garnet laser fusion of endarterectomy flaps.

    PubMed

    Humphrey, P W; Slocum, M M; Loy, T S; Silver, D

    1995-07-01

    This study evaluated the efficacy of neodymium:yttrium-aluminum-garnet laser welding of flaps in canine arteries and in securing the distal flap during human carotid endarterectomy. Endarterectomy flaps were created in both common carotid and both common femoral arteries in 12 dogs. The flaps were repaired with either the neodymium:yttrium-aluminum-garnet laser or with 6-0 polypropylene sutures. The arteries were removed after duplex scanning at either 7 or 28 days. Eighteen high carotid endarterectomy flaps in 16 patients have been subsequently secured with the laser welding technique. Laser repairs (125 +/- 19 joule) of the canine arteries were completed more quickly than suture repairs (mean 25 seconds vs 135 seconds, respectively; p < 0.04). Duplex ultrasonography revealed no discernable differences between the two groups of arteries. Arteries studied at 7 days revealed three microscopic flaps (two suture, one laser), more subintimal fibroblastic proliferation in suture than laser-repaired carotid arteries (3: 1, p = 0.0530), and similar amounts of inflammation in suture- and laser-repaired arteries. Arteries studied at 28 days revealed one microscopic intimal flap (suture-repaired); equal fibroblastic and inflammatory responses in suture- and laser-repaired vessels; and no evidence of laser thermal injury. Eighteen carotid endarterectomy flaps have been successfully fused with no immediate or long-term complications in 16 patients (follow-up of 0 to 24 months). Laser fusion appears to be a safe and effective method for securing distal carotid endarterectomy flaps.

  5. Nanosurgery with near-infrared 12-femtosecond and picosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Zhang, Huijing; Lemke, Cornelius; König, Karsten

    2011-03-01

    Laser-assisted surgery based on multiphoton absorption of NIR laser light has great potential for high precision surgery at various depths within the cells and tissues. Clinical applications include refractive surgery (fs-LASIK). The non-contact laser method also supports contamination-free cell nanosurgery. Here we apply femtosecond laser scanning microscopes for sub-100 nm surgery of human cells and metaphase chromosomes. A mode-locked 85 MHz Ti:Sapphire laser with an M-shaped ultrabroad band spectrum (maxima: 770 nm/830 nm) with an in situ pulse duration at the target ranging from 12 femtoseconds up to 3 picoseconds was employed. The effects of laser nanoprocessing in cells and chromosomes have been quantified by atomic force microscopy (AFM) and electron microscopy. These studies demonstrate the potential of extreme ultrashort femtosecond laser pulses at low mean milliwatt powers for sub-100 nm surgery.

  6. Nanosurgery of cells and chromosomes using near-infrared twelve-femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, Aisada; Lessel, Matthias; Nietzsche, Sander; Zeitz, Christian; Jacobs, Karin; Lemke, Cornelius; König, Karsten

    2012-10-01

    Laser-assisted surgery based on multiphoton absorption of near-infrared laser light has great potential for high precision surgery at various depths within the cells and tissues. Clinical applications include refractive surgery (fs-LASIK). The non-contact laser method also supports contamination-free cell nanosurgery. In this paper we describe usage of an ultrashort femtosecond laser scanning microscope for sub-100 nm surgery of human cells and metaphase chromosomes. A mode-locked 85 MHz Ti:Sapphire laser with an M-shaped ultrabroad band spectrum (maxima: 770 nm/830 nm) and an in situ pulse duration at the target ranging from 12 fs up to 3 ps was employed. The effects of laser nanoprocessing in cells and chromosomes have been quantified by atomic force microscopy. These studies demonstrate the potential of extreme ultrashort femtosecond laser pulses at low mean milliwatt powers for sub-100 nm surgery of cells and cellular organelles.

  7. Evaluating the use of laser radiation in cleaning of copper embroidery threads on archaeological Egyptian textiles

    NASA Astrophysics Data System (ADS)

    Abdel-Kareem, Omar; Harith, M. A.

    2008-07-01

    Cleaning of copper embroidery threads on archaeological textiles is still a complicated conservation process, as most textile conservators believe that the advantages of using traditional cleaning techniques are less than their disadvantages. In this study, the uses of laser cleaning method and two modified recipes of wet cleaning methods were evaluated for cleaning of the corroded archaeological Egyptian copper embroidery threads on an archaeological Egyptian textile fabric. Some corroded copper thread samples were cleaned using modified recipes of wet cleaning method; other corroded copper thread samples were cleaned with Q-switched Nd:YAG laser radiation of wavelength 532 nm. All tested metal thread samples before and after cleaning were investigated using a light microscope and a scanning electron microscope with an energy dispersive X-ray analysis unit. Also the laser-induced breakdown spectroscopy (LIBS) technique was used for the elemental analysis of laser-cleaned samples to follow up the laser cleaning procedure. The results show that laser cleaning is the most effective method among all tested methods in the cleaning of corroded copper threads. It can be used safely in removing the corrosion products without any damage to both metal strips and fibrous core. The tested laser cleaning technique has solved the problems caused by other traditional cleaning techniques that are commonly used in the cleaning of metal threads on museum textiles.

  8. Single step high-speed printing of continuous silver lines by laser-induced forward transfer

    NASA Astrophysics Data System (ADS)

    Puerto, D.; Biver, E.; Alloncle, A.-P.; Delaporte, Ph.

    2016-06-01

    The development of high-speed ink printing process by Laser-Induced Forward Transfer (LIFT) is of great interest for the printing community. To address the problems and the limitations of this process that have been previously identified, we have performed an experimental study on laser micro-printing of silver nanoparticle inks by LIFT and demonstrated for the first time the printing of continuous conductive lines in a single pass at velocities of 17 m/s using a 1 MHz repetition rate laser. We investigated the printing process by means of a time-resolved imaging technique to visualize the ejection dynamics of single and adjacent jets. The control of the donor film properties is of prime importance to achieve single step printing of continuous lines at high velocities. We use a 30 ps pulse duration laser with a wavelength of 343 nm and a repetition rate from 0.2 to 1 MHz. A galvanometric mirror head controls the distance between two consecutives jets by scanning the focused beam along an ink-coated donor substrate at different velocities. Droplets and lines of silver inks are laser-printed on glass and PET flexible substrates and we characterized their morphological quality by atomic force microscope (AFM) and optical microscope.

  9. Direct graphene growth from highly ordered pyrolytic graphite using pulsed Nd: YAG laser on p-Si (100) substrate at 700°c

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

    Kumar, Pramod, E-mail: kumarpramod.iitd@gmail.com; Lahiri, Indranil; Mitra, Anirban

    Few layer graphene was deposited on p-type Si (100) substrates by pulsed laser deposition of highly ordered pyrolytic graphite (HOPG) target at a relatively low temperature of 700 °C, without any catalytic layer. Effect of laser energy on the ability to produce the crystalline graphene was studied. It was observed that a laser energy of 220 mJ/pulse lead to form few layer graphene while higher laser energy of 440 mJ/pulse was detrimental to precipitation process. The reasons behind this observation are also discussed. Graphene samples were analyzed using Raman spectroscopy and surface morphology of graphene samples was confirmed using fieldmore » emission scanning electron microscope (FE-SEM).« less

  10. Evaluation Of The Shear Bond Strength Between Dentin And Dental Luting Cement Following Dentin Surface Treatment By 980 Nm Diode Laser And Desensitizing Agent

    NASA Astrophysics Data System (ADS)

    Ibrahim, T.; Gheith, M.

    2011-09-01

    Dentin hypersensitivity is described clinically as an exaggerated response to non-noxious sensory stimuli. Current treatment is concentrating on two approaches; to occlude the dentinal tubules or to block neural transmission. This is achieved through using dentin desensitizers and low power lasers. Forty eight freshly extracted human molar teeth were used in this study and divided equally into three groups. Group 1) control group, group 2) laser treated dentin surface group, and group 3) desensitizing agent dentin surface group. Scanning electron microscopic analysis of laser treated group showed melted globules, no carbonization, recrystalization and crystal growth of the apatite in some areas. In diode laser dentin surface treated group showed the highest shear bond strength mean value.

  11. Negatively-chirped laser enables nonlinear excitation and nanoprocessing with sub-20-fs pulses

    NASA Astrophysics Data System (ADS)

    Uchugonova, A.; Müller, J.; Bückle, R.; Tempea, G.; Isemann, A.; Stingl, A.; König, K.

    2008-02-01

    It has long been considered that the advantages emerging from employing chirp pre-compensation in nonlinear microscopy were overweighed by the complexity of prism- or grating-based compressors. These concerns were refuted with the advent of dispersive-mirrors-based compressors that are compact, user-friendly and sufficiently accurate to support sub-20-fs pulse delivery. Recent advances in the design of dispersive multilayer mirrors resulted in improved bandwidth (covering now as much as half of the gain bandwidth of Ti:Sapphire) and increased dispersion per bounce (one reflection off a state-of-the-art dispersive mirror pre-compensates the dispersion corresponding to >10mm of glass). The compressor built with these mirrors is sufficiently compact to be integrated in the housing of a sub-12-fs Ti:Sapphire oscillator. A complete scanning nonlinear microscope (FemtOgene, JenLab GmbH) equipped with highly-dispersive, large-NA objectives (Zeiss EC Plan-Neofluoar 40x/1.3, Plan-Neofluar 63x/1,25 Oil) was directly seeded with this negatively chirped laser. The pulse duration was measured at the focus of the objectives by inserting a scanning autocorrelator in the beam path between the laser and the microscope and recording the second order interferometric autocorrelation traces with the detector integrated in the microscope. Pulse durations <20fs were measured with both objectives. The system has been applied for two-photon imaging, transfection and optical manipulation of stem cells. Here we report on the successful transfection of human stem cells by transient optoporation of the cell membrane with a low mean power of < 7 mW and a short μs beam dwell time. Optically transfected cells were able to reproduce. The daughter cell expressed also green fluorescent proteins (GFP) indicating the successful modification of the cellular DNA.

  12. Ultrastructure observation of middle ear mucosa with laser irradiation

    NASA Astrophysics Data System (ADS)

    Kang, Mengkui; Yang, Shulan; Fang, Yaoyun; Sun, Jianhe

    1998-08-01

    In order to study the effects of He-Ne laser on the mucosa of middle ear mucosa from 9 patients with chronic otitis media, all of who had slight damp eardrum, were irradiated by low power He-Ne laser ten minutes per day for ten days. Specimen was taken before and after irradiation and observed under scanning electron microscope. It was found that the surface structure of the mucosa was more integral, the arrangement of the epithelial cell was closer together and microvilli arose among the noncilliated cells after irradiation. The inflammatory cell disappeared arid the morphologic structure appeared normal. These data provided the therapeutic evidence for the lower power He-Ne laser irradiation on patients with chronic purulent otitis midia.

  13. Accurate potential drop sheet resistance measurements of laser-doped areas in semiconductors

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

    Heinrich, Martin, E-mail: mh.seris@gmail.com; NUS Graduate School for Integrative Science and Engineering, National University of Singapore, Singapore 117456; Kluska, Sven

    2014-10-07

    It is investigated how potential drop sheet resistance measurements of areas formed by laser-assisted doping in crystalline Si wafers are affected by typically occurring experimental factors like sample size, inhomogeneities, surface roughness, or coatings. Measurements are obtained with a collinear four point probe setup and a modified transfer length measurement setup to measure sheet resistances of laser-doped lines. Inhomogeneities in doping depth are observed from scanning electron microscope images and electron beam induced current measurements. It is observed that influences from sample size, inhomogeneities, surface roughness, and coatings can be neglected if certain preconditions are met. Guidelines are given onmore » how to obtain accurate potential drop sheet resistance measurements on laser-doped regions.« less

  14. Near-field microscopy of waveguide architectures of InGaN/GaN diode lasers

    NASA Astrophysics Data System (ADS)

    Friede, Sebastian; Tomm, Jens W.; Kühn, Sergei; Hoffmann, Veit; Wenzel, Hans; Weyers, Markus

    2016-11-01

    Waveguide (WG) architectures of 420 nm emitting InGaN/GaN diode lasers are analyzed by photoluminescence and photocurrent spectroscopy using a nearfield scanning optical microscope that scans along their front facets. The components of the ‘optical active cavity’, quantum wells, WGs, and cladding layers are individually inspected with a spatial resolution of ∼100 nm. Separate analysis of the p- and n-sections of the WG was achieved, and reveals defect levels in the p-part. Moreover, it is demonstrated that the homogeneity of the n-WG section directly affects the quantum wells that are grown on top of this layer. Substantially increased carrier capture efficiencies into InGaN/GaN-WGs compared to GaN-WGs are demonstrated.

  15. Multimodal ophthalmic imaging using spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography

    NASA Astrophysics Data System (ADS)

    El-Haddad, Mohamed T.; Malone, Joseph D.; Li, Jianwei D.; Bozic, Ivan; Arquitola, Amber M.; Joos, Karen M.; Patel, Shriji N.; Tao, Yuankai K.

    2017-08-01

    Ophthalmic surgery involves manipulation of delicate, layered tissue structures on milli- to micrometer scales. Traditional surgical microscopes provide an inherently two-dimensional view of the surgical field with limited depth perception which precludes accurate depth-resolved visualization of these tissue layers, and limits the development of novel surgical techniques. We demonstrate multimodal swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography (SS-SESLO-OCT) to address current limitations of image-guided ophthalmic microsurgery. SS-SESLO-OCT provides inherently co-registered en face and cross-sectional field-of-views (FOVs) at a line rate of 400 kHz and >2 GPix/s throughput. We show in vivo imaging of the anterior segment and retinal fundus of a healthy volunteer, and preliminary results of multi-volumetric mosaicking for ultrawide-field retinal imaging with 90° FOV. Additionally, a scan-head was rapid-prototyped with a modular architecture which enabled integration of SS-SESLO-OCT with traditional surgical microscope and slit-lamp imaging optics. Ex vivo surgical maneuvers were simulated in cadaveric porcine eyes. The system throughput enabled volumetric acquisition at 10 volumes-per-second (vps) and allowed visualization of surgical dynamics in corneal sweeps, compressions, and dissections, and retinal sweeps, compressions, and elevations. SESLO en face images enabled simple real-time co-registration with the surgical microscope FOV, and OCT cross-sections provided depth-resolved visualization of instrument-tissue interactions. Finally, we demonstrate novel augmented-reality integration with the surgical view using segmentation overlays to aid surgical guidance. SS-SESLO-OCT may benefit clinical diagnostics by enabling aiming, registration, and mosaicking; and intraoperative imaging by allowing for real-time surgical feedback, instrument tracking, and overlays of computationally extracted biomarkers of disease.

  16. Frequency dependence of tissue attenuation measured by acoustic microscopy.

    PubMed

    Daft, C M; Briggs, G A; O'Brien, W D

    1989-05-01

    Broadband scanning acoustic microscopy (SAM) has been used to investigate the mechanical properties of sections of tissue with a resolution of around 8 microns. The work reported here extends these results by reporting the frequency dependence of the attenuation coefficient from 100-500 MHz. A discussion of the theory of the measurements is presented. The scanning laser acoustic microscope (SLAM) is used to characterize similar tissue sections at 100 MHz. The data obtained with the two forms of acoustic microscopy are compared with results from the literature.

  17. Numerical simulation of dendrite growth in nickel-based superalloy and validated by in-situ observation using high temperature confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Yan, Xuewei; Xu, Qingyan; Liu, Baicheng

    2017-12-01

    Dendritic structures are the predominant microstructural constituents of nickel-based superalloys, an understanding of the dendrite growth is required in order to obtain the desirable microstructure and improve the performance of castings. For this reason, numerical simulation method and an in-situ observation technology by employing high temperature confocal laser scanning microscopy (HT-CLSM) were used to investigate dendrite growth during solidification process. A combined cellular automaton-finite difference (CA-FD) model allowing for the prediction of dendrite growth of binary alloys was developed. The algorithm of cells capture was modified, and a deterministic cellular automaton (DCA) model was proposed to describe neighborhood tracking. The dendrite and detail morphology, especially hundreds of dendrites distribution at a large scale and three-dimensional (3-D) polycrystalline growth, were successfully simulated based on this model. The dendritic morphologies of samples before and after HT-CLSM were both observed by optical microscope (OM) and scanning electron microscope (SEM). The experimental observations presented a reasonable agreement with the simulation results. It was also found that primary or secondary dendrite arm spacing, and segregation pattern were significantly influenced by dendrite growth. Furthermore, the directional solidification (DS) dendritic evolution behavior and detail morphology were also simulated based on the proposed model, and the simulation results also agree well with experimental results.

  18. Electrically tunable lens speeds up 3D orbital tracking

    PubMed Central

    Annibale, Paolo; Dvornikov, Alexander; Gratton, Enrico

    2015-01-01

    3D orbital particle tracking is a versatile and effective microscopy technique that allows following fast moving fluorescent objects within living cells and reconstructing complex 3D shapes using laser scanning microscopes. We demonstrated notable improvements in the range, speed and accuracy of 3D orbital particle tracking by replacing commonly used piezoelectric stages with Electrically Tunable Lens (ETL) that eliminates mechanical movement of objective lenses. This allowed tracking and reconstructing shape of structures extending 500 microns in the axial direction. Using the ETL, we tracked at high speed fluorescently labeled genomic loci within the nucleus of living cells with unprecedented temporal resolution of 8ms using a 1.42NA oil-immersion objective. The presented technology is cost effective and allows easy upgrade of scanning microscopes for fast 3D orbital tracking. PMID:26114037

  19. Electron beam assisted field evaporation of insulating nanowires/tubes

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

    Blanchard, N. P., E-mail: nicholas.blanchard@univ-lyon1.fr; Niguès, A.; Choueib, M.

    2015-05-11

    We demonstrate field evaporation of insulating materials, specifically BN nanotubes and undoped Si nanowires, assisted by a convergent electron beam. Electron irradiation leads to positive charging at the nano-object's apex and to an important increase of the local electric field thus inducing field evaporation. Experiments performed both in a transmission electron microscope and in a scanning electron microscope are presented. This technique permits the selective evaporation of individual nanowires in complex materials. Electron assisted field evaporation could be an interesting alternative or complementary to laser induced field desorption used in atom probe tomography of insulating materials.

  20. In Situ Observations of Agglomeration of Non-metallic Inclusions at Steel/Ar and Steel/Slag Interfaces by High-Temperature Confocal Laser Scanning Microscope: A Review

    NASA Astrophysics Data System (ADS)

    Mu, Wangzhong; Dogan, Neslihan; Coley, Kenneth S.

    2018-05-01

    The agglomeration behavior of non-metallic inclusions in the steelmaking process is important for controlling the cleanliness of the steel. In this work, the observation of agglomeration behaviors of inclusions at steel/Ar and steel/slag interfaces using a high-temperature confocal laser scanning microscope (HT-CLSM) is summarized. This HT-CLSM technique has been applied to observe phase transformation during solidification and heat treatment and the engulfment and pushing behavior of inclusions in front of the solidified interface. In the current work, the inclusion agglomeration behavior at steel/Ar and steel/slag interfaces is summarized and discussed. Subsequently, the development of the theoretical work investigating inclusion agglomeration at steel/Ar and steel/slag interfaces including the initial capillary force model and Kralchevsky-Paunov model is described. Finally, the Kralchevsky-Paunov model is applied to investigating nitride inclusion agglomeration at high-manganese steel/Ar interfaces. This work aims to give a critical review of the application of HT-CLSM in secondary refining as well as a better control of inclusion elimination for clean steel production.

  1. Confocal laser scanning microscopy in study of bone calcification

    NASA Astrophysics Data System (ADS)

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

    2012-12-01

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

  2. Crystallization Behavior of the CaO-Al2O3-MgO System Studied with a Confocal Laser Scanning Microscope

    NASA Astrophysics Data System (ADS)

    Jung, Sung Suk; Sohn, Il

    2012-12-01

    The crystallization behavior of a calcium-aluminate system with various MgO content from 2.5 to 7.5 wt pct and CaO/Al2O3 ratios between 0.8 and 1.2 has been examined using a confocal laser scanning microscope (CLSM). CCT (continuous cooling transformation) and time temperature transformation (TTT) diagrams were constructed to identify the primary crystal phase of slag at different compositions and at cooling rates between 25 and 800 K/minutes. In the slag at a CaO/Al2O3 ratio of 1.0, crystallization temperature increased during isothermal and continuous cooling with higher MgO content, and the shortest incubation time was observed at 5 wt pct MgO. When MgO content was fixed to be 5 wt pct, crystallization temperature increased with lower CaO/Al2O3 ratio. According to the slag composition, cooling rates and temperature, the primary phase could be CA, or C5A3, or C3A, or C3MA2, or MgO, and the crystal morphology changes from dendrites to faceted crystals to columnar crystals in this composition range.

  3. Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices

    PubMed Central

    Losavio, Bradley E.; Iyer, Vijay; Saggau, Peter

    2009-01-01

    We developed a two-photon microscope optimized for physiologically manipulating single neurons through their postsynaptic receptors. The optical layout fulfills the stringent design criteria required for high-speed, high-resolution imaging in scattering brain tissue with minimal photodamage. We detail the practical compensation of spectral and temporal dispersion inherent in fast laser beam scanning with acousto-optic deflectors, as well as a set of biological protocols for visualizing nearly diffraction-limited structures and delivering physiological synaptic stimuli. The microscope clearly resolves dendritic spines and evokes electrophysiological transients in single neurons that are similar to endogenous responses. This system enables the study of multisynaptic integration and will assist our understanding of single neuron function and dendritic computation. PMID:20059271

  4. A compact light-sheet microscope for the study of the mammalian central nervous system

    PubMed Central

    Yang, Zhengyi; Haslehurst, Peter; Scott, Suzanne; Emptage, Nigel; Dholakia, Kishan

    2016-01-01

    Investigation of the transient processes integral to neuronal function demands rapid and high-resolution imaging techniques over a large field of view, which cannot be achieved with conventional scanning microscopes. Here we describe a compact light sheet fluorescence microscope, featuring a 45° inverted geometry and an integrated photolysis laser, that is optimized for applications in neuroscience, in particular fast imaging of sub-neuronal structures in mammalian brain slices. We demonstrate the utility of this design for three-dimensional morphological reconstruction, activation of a single synapse with localized photolysis, and fast imaging of neuronal Ca2+ signalling across a large field of view. The developed system opens up a host of novel applications for the neuroscience community. PMID:27215692

  5. One-step synthesis of hybrid inorganic-organic nanocomposite coatings by novel laser adaptive ablation deposition technique

    NASA Astrophysics Data System (ADS)

    Serbezov, Valery; Sotirov, Sotir

    2013-03-01

    A novel approach for one-step synthesis of hybrid inorganic-organic nanocomposite coatings by new modification of Pulsed Laser Deposition technology called Laser Adaptive Ablation Deposition (LAAD) is presented. Hybrid nanocomposite coatings including Mg- Rapamycin and Mg- Desoximetasone were produced by UV TEA N2 laser under low vacuum (0.1 Pa) and room temperature onto substrates from SS 316L, KCl and NaCl. The laser fluence for Mg alloy was 1, 8 J/cm2 and for Desoximetasone 0,176 J/cm2 and for Rapamycin 0,118 J/cm2 were respectively. The threedimensional two-segmented single target was used to adapt the interaction of focused laser beam with inorganic and organic material. Magnesium alloy nanoparticles with sizes from 50 nm to 250 nm were obtained in organic matrices. The morphology of nanocomposites films were studied by Bright field / Fluorescence optical microscope and Scanning Electron Microscope (SEM). Fourier Transform Infrared (FTIR) spectroscopy measurements were applied in order to study the functional properties of organic component before and after the LAAD process. Energy Dispersive X-ray Spectroscopy (EDX) was used for identification of Mg alloy presence in hybrid nanocomposites coatings. The precise control of process parameters and particularly of the laser fluence adjustment enables transfer on materials with different physical chemical properties and one-step synthesis of complex inorganic- organic nanocomposites coatings.

  6. 3-D reconstruction of neurons from multichannel confocal laser scanning image series.

    PubMed

    Wouterlood, Floris G

    2014-04-10

    A confocal laser scanning microscope (CLSM) collects information from a thin, focal plane and ignores out-of-focus information. Scanning of a specimen, with stepwise axial (Z-) movement of the stage in between each scan, produces Z-series of confocal images of a tissue volume, which then can be used to 3-D reconstruct structures of interest. The operator first configures separate channels (e.g., laser, filters, and detector settings) for each applied fluorochrome and then acquires Z-series of confocal images: one series per channel. Channel signal separation is extremely important. Measures to avoid bleaching are vital. Post-acquisition deconvolution of the image series is often performed to increase resolution before 3-D reconstruction takes place. In the 3-D reconstruction programs described in this unit, reconstructions can be inspected in real time from any viewing angle. By altering viewing angles and by switching channels off and on, the spatial relationships of 3-D-reconstructed structures with respect to structures visualized in other channels can be studied. Since each brand of CLSM, computer program, and 3-D reconstruction package has its own proprietary set of procedures, a general approach is provided in this protocol wherever possible. Copyright © 2014 John Wiley & Sons, Inc.

  7. Coherent anti-Stokes Raman scattering microscope with a high-signal-to-noise ratio, high stability, and high-speed imaging for live cell observation

    NASA Astrophysics Data System (ADS)

    Hayashi, Shinichi; Takimoto, Shinichi; Hashimoto, Takeshi

    2007-02-01

    Coherent anti-Stokes Raman scattering (CARS) microscopy, which can produce images of specific molecules without staining, has attracted the attention of researchers, as it matches the need for molecular imaging and pathway analysis of live cells. In particular, there have been an increasing number of CARS experimental results regarding lipids in live cells, which cannot be fluorescently tagged while keeping the cells alive. One of the important applications of lipid research is for the metabolic syndrome. Since the metabolic syndrome is said to be related to the lipids in lipocytes, blood, arterial vessels, and so on, the CARS technique is expected to find application in this field. However, CARS microscopy requires a pair of picosecond laser pulses, which overlap both temporally and spatially. This makes the optical adjustments of a CARS microscope challenging. The authors developed a CARS unit that includes optics for easy and stable adjustment of the overlap of these laser pulses. Adding the CARS unit to a laser scanning microscope provides CARS images of a high signal-to-noise ratio, with an acquisition rate as high as 2 microseconds per pixel. Thus, images of fast-moving lipid droplets in Hela cells were obtained.

  8. Efficacy of 4 Irrigation Protocols in Killing Bacteria Colonized in Dentinal Tubules Examined by a Novel Confocal Laser Scanning Microscope Analysis

    PubMed Central

    Azim, Adham A.; Aksel, Hacer; Zhuang, Tingting; Mashtare, Terry; Babu, Jegdish P.; Huang, George T.-J.

    2016-01-01

    Introduction The aim of this study was to determine the efficiency of 4 irrigation systems in eliminating bacteria in root canals, particularly in dentinal tubules. Methods Roots of human teeth were prepared to 25/04, autoclaved, and inoculated with Enterococcus faecalis for 3 weeks. Canals were then disinfected by (1) standard needle irrigation, (2) sonically agitating with EndoActivator, (3) XP Endo finisher, or (4) erbium:yttrium aluminum garnet laser (PIPS) (15 roots/group). The bacterial reduction in the canal was determined by MTT assays. For measuring live versus dead bacteria in the dentinal tubules (4 teeth/group), teeth were split open and stained with LIVE/DEAD BackLight. Coronal, middle, and apical thirds of the canal dentin were scanned by using a confocal laser scanning microscope (CLSM) to determine the ratio of dead/total bacteria in the dentinal tubules at various depths. Results All 4 irrigation protocols significantly eliminated bacteria in the canal, ranging from 89.6% to 98.2% reduction (P < .001). XP Endo had the greatest bacterial reduction compared with other 3 techniques (P < .05). CLSM analysis showed that XP Endo had the highest level of dead bacteria in the coronal, middle, and apical segments at 50-μm depth. On the other hand, PIPS had the greatest bacterial killing efficiency at the 150-μm depth in all 3 root segments. Conclusions XP Endo appears to be more efficient than other 3 techniques in disinfecting the main canal space and up to 50 μm deep into the dentinal tubules. PIPS appears to be most effective in killing the bacteria deep in the dentinal tubules. PMID:27130334

  9. Efficacy of 4 Irrigation Protocols in Killing Bacteria Colonized in Dentinal Tubules Examined by a Novel Confocal Laser Scanning Microscope Analysis.

    PubMed

    Azim, Adham A; Aksel, Hacer; Zhuang, Tingting; Mashtare, Terry; Babu, Jegdish P; Huang, George T-J

    2016-06-01

    The aim of this study was to determine the efficiency of 4 irrigation systems in eliminating bacteria in root canals, particularly in dentinal tubules. Roots of human teeth were prepared to 25/04, autoclaved, and inoculated with Enterococcus faecalis for 3 weeks. Canals were then disinfected by (1) standard needle irrigation, (2) sonically agitating with EndoActivator, (3) XP Endo finisher, or (4) erbium:yttrium aluminum garnet laser (PIPS) (15 roots/group). The bacterial reduction in the canal was determined by MTT assays. For measuring live versus dead bacteria in the dentinal tubules (4 teeth/group), teeth were split open and stained with LIVE/DEAD BackLight. Coronal, middle, and apical thirds of the canal dentin were scanned by using a confocal laser scanning microscope (CLSM) to determine the ratio of dead/total bacteria in the dentinal tubules at various depths. All 4 irrigation protocols significantly eliminated bacteria in the canal, ranging from 89.6% to 98.2% reduction (P < .001). XP Endo had the greatest bacterial reduction compared with other 3 techniques (P < .05). CLSM analysis showed that XP Endo had the highest level of dead bacteria in the coronal, middle, and apical segments at 50-μm depth. On the other hand, PIPS had the greatest bacterial killing efficiency at the 150-μm depth in all 3 root segments. XP Endo appears to be more efficient than other 3 techniques in disinfecting the main canal space and up to 50 μm deep into the dentinal tubules. PIPS appears to be most effective in killing the bacteria deep in the dentinal tubules. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  10. Laser cutting of Kevlar laminates and thermal stress formed at cutting sections

    NASA Astrophysics Data System (ADS)

    Yilbas, B. S.; Akhtar, S. S.

    2012-02-01

    Laser cutting of Kevlar laminates is carried out and thermal stress field developed in the cutting region is predicted using the finite element code. Temperature predictions are validated through the thermocouple data. The morphological changes in the cutting section are examined by incorporating optical and scanning electron microscopes. It is found that temperature predictions agree well with the thermocouple data. High values of von Mises stress are observed at the cutting edges and at the mid-thickness of the Kevlar laminate due to thermal compression formed in this region. The laser cut edges are free from whiskers; however, striation formation and some small sideways burning is observed at the kerf edges.

  11. Confocal Laser Scanning Microscopy, a New In Vivo Diagnostic Tool for Schistosomiasis

    PubMed Central

    Holtfreter, Martha Charlotte; Nohr-Łuczak, Constanze; Guthoff, Rudolf Friedrich; Reisinger, Emil Christian

    2012-01-01

    Background The gold standard for the diagnosis of schistosomiasis is the detection of the parasite's characteristic eggs in urine, stool, or rectal and bladder biopsy specimens. Direct detection of eggs is difficult and not always possible in patients with low egg-shedding rates. Confocal laser scanning microscopy (CLSM) permits non-invasive cell imaging in vivo and is an established way of obtaining high-resolution images and 3-dimensional reconstructions. Recently, CLSM was shown to be a suitable method to visualize Schistosoma mansoni eggs within the mucosa of dissected mouse gut. In this case, we evaluated the suitability of CLSM to detect eggs of Schistosoma haematobium in a patient with urinary schistosomiasis and low egg-shedding rates. Methodology/Principal Findings The confocal laser scanning microscope used in this study was based on a scanning laser system for imaging the retina of a living eye, the Heidelberg Retina Tomograph II, in combination with a lens system (image modality). Standard light cystoscopy was performed using a rigid cystoscope under general anaesthesia. The CLSM endoscope was then passed through the working channel of the rigid cystoscope. The mucosal tissue of the bladder was scanned using CLSM. Schistoma haematobium eggs appeared as bright structures, with the characteristic egg shape and typical terminal spine. Conclusion/Significance We were able to detect schistosomal eggs in the urothelium of a patient with urinary schistosomiasis. Thus, CLSM may be a suitable tool for the diagnosis of schistosomiasis in humans, especially in cases where standard diagnostic tools are not suitable. PMID:22529947

  12. Resolution enhancement of pump-probe microscope with an inverse-annular filter

    NASA Astrophysics Data System (ADS)

    Kobayashi, Takayoshi; Kawasumi, Koshi; Miyazaki, Jun; Nakata, Kazuaki

    2018-04-01

    Optical pump-probe microscopy can provide images by detecting changes in probe light intensity induced by stimulated emission, photoinduced absorbance change, or photothermal-induced refractive index change in either transmission or reflection mode. Photothermal microscopy, which is one type of optical pump-probe microscopy, has intrinsically super resolution capability due to the bilinear dependence of signal intensity of pump and probe. We introduce new techniques for further resolution enhancement and fast imaging in photothermal microscope. First, we introduce a new pupil filter, an inverse-annular pupil filter in a pump-probe photothermal microscope, which provides resolution enhancement in three dimensions. The resolutions are proved to be improved in lateral and axial directions by imaging experiment using 20-nm gold nanoparticles. The improvement in X (perpendicular to the common pump and probe polarization direction), Y (parallel to the polarization direction), and Z (axial direction) are by 15 ± 6, 8 ± 8, and 21 ± 2% from the resolution without a pupil filter. The resolution enhancement is even better than the calculation using vector field, which predicts the corresponding enhancement of 11, 8, and 6%. The discussion is made to explain the unexpected results. We also demonstrate the photothermal imaging of thick biological samples (cells from rabbit intestine and kidney) stained with hematoxylin and eosin dye with the inverse-annular filter. Second, a fast, high-sensitivity photothermal microscope is developed by implementing a spatially segmented balanced detection scheme into a laser scanning microscope using a Galvano mirror. We confirm a 4.9 times improvement in signal-to-noise ratio in the spatially segmented balanced detection compared with that of conventional detection. The system demonstrates simultaneous bi-modal photothermal and confocal fluorescence imaging of transgenic mouse brain tissue with a pixel dwell time of 20 µs. The fluorescence image visualizes neurons expressing yellow fluorescence proteins, while the photothermal signal detected endogenous chromophores in the mouse brain, allowing 3D visualization of the distribution of various features such as blood cells and fine structures most probably due to lipids. This imaging modality was constructed using compact and cost-effective laser diodes, and will thus be widely useful in the life and medical sciences. Third, we have made further resolution improvement of high-sensitivity laser scanning photothermal microscopy by applying non-linear detection. By this, the new method has super resolution with 61 and 42% enhancement from the diffraction limit values of the probe and pump wavelengths, respectively, by a second-order non-linear scheme and a high-frame rate in a laser scanning microscope. The maximum resolution is determined to be 160 nm in the second-order non-linear detection mode and 270 nm in the linear detection mode by the PT signal of GNPs. The pixel rate and frame rate for 300 × 300 pixel image are 50 µs and 4.5 s, respectively. The pixel and frame rate are shorter than the rates, those are 1 ms and 100 s, using the piezo-driven stage system.

  13. A comparison of caveolae and caveolin-1 to folate receptor α in retina and retinal pigment epithelium

    PubMed Central

    Bridges, Christy C.; El-Sherbeny, Amira; Roon, Penny; Ola, M. Shamsul; Kekuda, Ramesh; Ganapathy, Vadivel; Cameron, Richard S.; Cameron, Patricia L.

    2015-01-01

    Summary Caveolae are flask-shaped membrane invaginations present in most mammalian cells. They are distinguished by the presence of a striated coat composed of the protein, caveolin. Caveolae have been implicated in numerous cellular processes, including potocytosis in which caveolae are hypothesized to co-localize with folate receptor α and participate in folate uptake. Our laboratory has recently localized folate receptor α to the basolateral surface of the retinal pigment epithelium (RPE). It is present also in many other cells of the retina. In the present study, we asked whether caveolae were present in the RPE, and if so, whether their pattern of distribution was similar to folate receptor α. We also examined the distribution pattern of caveolin-1, which can be a marker of caveolae. Extensive electron microscopical analysis revealed caveolae associated with endothelial cells. However, none were detected in intact or cultured RPE. Laser scanning confocal microscopical analysis of intact RPE localized caveolin-1 to the apical and basal surfaces, a distribution unlike folate receptor α. Western analysis confirmed the presence of caveolin-1 in cultured RPE cells and laser scanning confocal microscopy localized the protein to the basal plasma membrane of the RPE, a distribution like that of folate receptor α. This distribution was confirmed by electron microscopic immunolocalization. The lack of caveolae in the RPE suggests that these structures may not be essential for folate internalization in the RPE. PMID:11508338

  14. Laser micromilling of convex microfluidic channels onto glassy carbon for glass molding dies

    NASA Astrophysics Data System (ADS)

    Tseng, Shih-Feng; Chen, Ming-Fei; Hsiao, Wen-Tse; Huang, Chien-Yao; Yang, Chung-Heng; Chen, Yu-Sheng

    2014-06-01

    This study reports the fabrication of convex microfluidic channels on glassy carbon using an ultraviolet laser processing system to produce glass molding dies. The laser processing parameters, including various laser fluences and scanning speeds of galvanometers, were adjusted to mill a convex microchannel on a glassy carbon substrate to identify the effects of material removal. The machined glassy carbon substrate was then applied as a glass molding die to fabricate a glass-based microfluidic biochip. The surface morphology, milled width and depth, and surface roughness of the microchannel die after laser micromilling were examined using a three-dimensional confocal laser scanning microscope. This study also investigates the transcription rate of microchannels after the glass molding process. To produce a 180 μm high microchannel on the GC substrate, the optimal number of milled cycles, laser fluence, and scanning speed were 25, 4.9 J/cm2, and 200 mm/s, respectively. The width, height, and surface roughness of milled convex microchannels were 119.6±0.217 μm, 180.26±0.01 μm, and 0.672±0.08 μm, respectively. These measured values were close to the predicted values and suitable for a glass molding die. After the glass molding process, a typical glass-based microchannel chip was formed at a molding temperature of 660 °C and the molding force of 0.45 kN. The transcription rates of the microchannel width and depth were 100% and 99.6%, respectively. Thus, the proposed approach is suitable for performing in chemical, biochemical, or medical reactions.

  15. Bioactivity of calcium phosphate bioceramic coating fabricated by laser cladding

    NASA Astrophysics Data System (ADS)

    Zhu, Yizhi; Liu, Qibin; Xu, Peng; Li, Long; Jiang, Haibing; Bai, Yang

    2016-05-01

    There were always strong expectations for suitable biomaterials used for bone regeneration. In this study, to improve the biocompatiblity of titanium alloy, calcium phosphate bioceramic coating was obtained by laser cladding technology. The microstructure, phases, bioactivity, cell differentiation, morphology and resorption lacunae were investigated by optical microscope (OM), x-ray diffraction (XRD), methyl thiazolyl tetrazolium (MTT) assay, tartrate-resistant acid phosphatase (TRAP) staining and scanning electronic microscope (SEM), respectively. The results show that bioceramic coating consists of three layers, which are a substrate, an alloyed layer and a ceramic layer. Bioactive phases of β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) were found in ceramic coating. Osteoclast precursors have excellent proliferation on the bioceramic surface. The bioceramics coating could be digested by osteoclasts, which led to the resorption lacunae formed on its surface. It revealed that the gradient bioceramic coating has an excellent bioactivity.

  16. Three-photon fluorescence imaging of melanin with a dual-wedge confocal scanning system

    NASA Astrophysics Data System (ADS)

    Mega, Yair; Kerimo, Joseph; Robinson, Joseph; Vakili, Ali; Johnson, Nicolette; DiMarzio, Charles

    2012-03-01

    Confocal microscopy can be used as a practical tool in non-invasive applications in medical diagnostics and evaluation. In particular, it is being used for the early detection of skin cancer to identify pathological cellular components and, potentially, replace conventional biopsies. The detection of melanin and its spatial location and distribution plays a crucial role in the detection and evaluation of skin cancer. Our previous work has shown that the visible emission from melanin is strong and can be easily observed with a near-infrared CW laser using low power. This is due to a unique step-wise, (SW) three-photon excitation of melanin. This paper shows that the same SW, 3-photon fluorescence can also be achieved with an inexpensive, continuous-wave laser using a dual-prism scanning system. This demonstrates that the technology could be integrated into a portable confocal microscope for clinical applications. The results presented here are in agreement with images obtained with the larger and more expensive femtosecond laser system used earlier.

  17. Laser-induced damage of coatings on Yb:YAG crystals at cryogenic condition

    NASA Astrophysics Data System (ADS)

    Wang, He; Zhang, Weili; Chen, Shunli; Zhu, Meiping; He, Hongbo; Fan, Zhengxiu

    2011-12-01

    As large amounts of heat need to be dissipated during laser operation, some diode pumped solid state lasers (DPSSL), especially Yb:YAG laser, operate at cryogenic condition. This work investigated the laser induced damage of coatings (high-reflective and anti-reflective coatings) on Yb:YAG crystals at cryogenic temperature and room temperature. The results show that the damage threshold of coatings at cryogenic temperature is lower than the one at room temperature. Field-emission scanning electron microscopy (FESEM), optical profiler, step profiler and Atomic force microscope (AFM) were used to obtain the damage morphology, size and depth. Taking alteration of physical parameters, microstructure of coatings and the environmental pollution into consideration, we analyzed the key factor of lowering the coating damage threshold at cryogenic conditions. The results are important to understand the mechanisms leading to damage at cryogenic condition.

  18. Changes in surface morphology of enamel after Er:YAG laser irradiation

    NASA Astrophysics Data System (ADS)

    Rechmann, Peter; Goldin, Dan S.; Hennig, Thomas

    1998-04-01

    Aim of the study was to investigate the surface and subsurface structure of enamel after irradiation with an Er:YAG laser (wavelength 2.94 micrometer, pulse duration 250 - 500 microseconds, free running, beam profile close to tophead, focus diameter 600 micrometer, focus distance 13 mm, different power settings, air-water spray 2 ml/min; KAVO Key Laser 1242, Kavo Biberach, Germany). The surface of more than 40 freshly extracted wisdom teeth were irradiated using a standardized application protocol (pulse repetition rate 4 and 6 Hz, moving speed of the irradiation table 2 mm/sec and 3 mm/sec, respectively). On each surface between 3 and 5 tracks were irradiated at different laser energies (60 - 500 mJ/pulse) while each track was irradiated between one and ten times respectively. For the scanning electron microscope investigation teeth were dried in alcohol and sputtered with gold. For light microscopic examinations following laser impact, samples were fixed in formaldehyde, dried in alcohol and embedded in acrylic resin. Investigations revealed that at subsurface level cracks can not be observed even at application of highest energies. Borders of the irradiated tracks seem to be sharp while melted areas of different sizes are observed on the bottom of the tracks depending on applied energy. Small microcracks can be seen on the surface of these melted areas.

  19. Characteristics of plasma scalds in multilayer dielectric films

    NASA Astrophysics Data System (ADS)

    Liu, Xiaofeng; Zhao, Yuan'an; Li, Dawei; Hu, Guohang; Gao, Yanqi; Fan, Zhengxiu; Shao, Jianda

    2011-07-01

    Plasma scalding is one of the most typical laser damage morphologies induced by a nanosecond laser with a wavelength of 1053nm in HfO2/SiO2 multilayer films. In this paper, the characteristics of plasma scalds are systematically investigated with multiple methods. The scalding behaves as surface discoloration under a microscope. The shape is nearly circular when the laser incidence angle is close to normal incidence and is elliptical at oblique incidence. The nodular-ejection pit is in the center of the scalding region when the laser irradiates at the incidence angle close to normal incidence and in the right of the scalding region when the laser irradiates from left to right at oblique incidence. The maximum damage size of the scalding increases with laser energy. The edge of the scalding is high compared with the unirradiated film surface, and the region tending to the center is concave. Plasma scald is proved to be surface damage. The maximum depth of a scald increases with its size. Tiny pits of nanometer scale can be seen in the scalding film under a scanning electronic microscope at a higher magnification. The absorptions of the surface plasma scalds tend to be approximately the same as the lower absorptions of test sites without laser irradiation. Scalds do not grow during further illumination pulses until 65J/cm2. The formation of surface plasma scalding may be related to the occurrence of the laser-supported detonation wave.

  20. [Scanning electron microscopic investigations of cutting edge quality in lamellar keratotomy using the Wavelight femtosecond laser (FS-200) : What influence do spot distance and an additional tunnel have?

    PubMed

    Hammer, T; Höche, T; Heichel, J

    2018-01-01

    Femtosecond lasers (fs-lasers) are established cutting instruments for the creation of LASIK flaps. Previous studies often showed even rougher surfaces after application of fs-laser systems compared to lamellar keratotomy with mechanical microkeratomes. When cutting the cornea with fs-lasers, an intrastromal gas development occurs, which has a potentially negative influence on the cutting quality if the gas cannot be dissipated; therefore, manufacturers have chosen the way of gas assimilation in so-called pockets. The investigated system creates a tunnel which opens under the conjunctiva. The aim of this study was to investigate the effects of a tunnel as well as the influence of different spot distances on the quality of cut surfaces and edges. In this experimental study on freshly enucleated porcine eyes (n = 15), the following cuts were carried out with the FS-200 (Wavelight, Erlangen, Germany): 1. standard setting (spot and line separation 8 µm), 2. with tunnel for gas drainage, 3. without gas-conducting tunnel, 4. with increased spot spacing (spot and line separation 9 μm instead of 8 μm) and 5. with reduced spot spacing (spot and line separation 7 μm instead of 8 μm). Subsequently, scanning electron microscopy (FEI Quanta 650, Hillsboro, OR) of the cut edges and surfaces as well as the gas drain tunnel were performed. The evaluation was based on an established score. The current fs-laser system (200 Hz) is able to create smooth cutting surfaces and sharp edges. The changed density of laser pulses compared to the standard settings with a reduced or increased distance between the pulses, did not achieve any further improvement in the surface quality. The gas-conducting tunnel could be detected by scanning electron microscope. In the case of cutting without a tunnel, roughened surfaces and irregularities on the cutting edges were found. When the FS-200 fs-laser is used, LASIK cuts with very smooth cut surfaces and sharp cutting edges are achieved. This is only valid as long as an additional tunnel with the fs-laser is placed under the conjunctiva. It can be assumed that the resulting gas is effectively drained through this tunnel. The installation of the tunnel represents a new possibility to replace previous techniques of gas assimilation in deeper lying cutting areas.

  1. Simulation of vaporization in low fluence nanosecond laser ablation of aluminum alloy

    NASA Astrophysics Data System (ADS)

    Song, Chaoqun; Dong, Shiyun; Yan, Shixing; Li, Enzhong; Xu, Binshi; He, Peng

    2018-03-01

    This paper presents a multi-phase flow model for the nanosecond laser ablation of aluminum alloy at a low fluence based on finite volume method, considering gravity, recoil pressure, buoyancy and surface tension to describe vaporization. Actual morphology of ablation crater was measured by a laser scanning confocal microscope to verify the model. Results show that vaporization is the main ablation mechanism for 100ns laser ablation at low fluences, and the peak temperature is only 50% of critical temperature. Both the experimental and calculated crater have a wall-like bulge around the rim, as a result of impact of recoil pressure and resolidification of pushed liquid metal. The calculated depth and diameter of crater are in good agreement with the corresponding experimental measurement indicating the feasibility of the model.

  2. Fabrication and characterization of microcavity lasers in rhodamine B doped SU8 using high energy proton beam

    NASA Astrophysics Data System (ADS)

    Venugopal Rao, S.; Bettiol, A. A.; Vishnubhatla, K. C.; Bhaktha, S. N. B.; Narayana Rao, D.; Watt, F.

    2007-03-01

    The authors present their results on the characterization of individual dye-doped microcavity polymer lasers fabricated using a high energy proton beam. The lasers were fabricated in rhodamine B doped SU8 resist with a single exposure step followed by chemical processing. The resulting trapezoidal shaped cavities had dimensions of ˜250×250μm2. Physical characterization of these structures was performed using a scanning electron microscope while the optical characterization was carried out by recording the emission subsequent to pumping the lasers with 532nm, 6 nanosecond pulses. The authors observed intense, narrow emission near 624nm with the best emission linewidth full width at half maximum of ˜9nm and a threshold ˜150μJ/mm2.

  3. Cell optoporation with a sub-15 fs and a 250-fs laser

    NASA Astrophysics Data System (ADS)

    Breunig, Hans Georg; Batista, Ana; Uchugonova, Aisada; König, Karsten

    2016-06-01

    We employed two commercially available femtosecond lasers, a Ti:sapphire and a ytterbium-based oscillator, to directly compare from a user's practical point-of-view in one common experimental setup the efficiencies of transient laser-induced cell membrane permeabilization, i.e., of so-called optoporation. The experimental setup consisted of a modified multiphoton laser-scanning microscope employing high-NA focusing optics. An automatic cell irradiation procedure was realized with custom-made software that identified cell positions and controlled relevant hardware components. The Ti:sapphire and ytterbium-based oscillators generated broadband sub-15-fs pulses around 800 nm and 250-fs pulses at 1044 nm, respectively. A higher optoporation rate and posttreatment viability were observed for the shorter fs pulses, confirming the importance of multiphoton effects for efficient optoporation.

  4. Friction and fretting wear characteristics of different diamond-like carbon coatings against alumina in water-lubricated fretting conditions.

    PubMed

    Watabe, Tsukasa; Amanov, Auezhan; Tsuboi, Ryo; Sasaki, Shinya

    2013-12-01

    Diamond-like carbon (DLC) coatings typically show low friction and high wear resistance. In this study, the friction and fretting wear characteristics of PVD, CVD and CVD-Si DLC coatings were investigated against an alumina (Al2O3) ball under water-lubricated fretting conditions. The objective of this study is to investigate and compare the friction and fretting wear characteristics of those DLC coatings at various fretting frequencies. The test results showed that the PVD DLC coating led to a lower friction coefficient and a higher resistance to fretting wear compared to those of the CVD and CVD-Si DLC coatings. However, the CVD DLC coating showed that the fretting wear resistance decreases with increasing frequency, while no significant difference in fretting wear resistances of the PVD and CVD-Si DLC coatings was observed. Quantitative surface analyses of the specimens were performed using an energy dispersive spectroscopy (EDS), a laser scanning microscope (LSM), a scanning electron microscope (SEM), an atomic force microscope (AFM) and the Raman spectroscopy.

  5. Optical, structural and nonlinear optical properties of laser ablation synthesized Ag nanoparticles and photopolymer nanocomposites based on them

    NASA Astrophysics Data System (ADS)

    Zulina, Natalia A.; Pavlovetc, Ilia M.; Baranov, Mikhail A.; Denisyuk, Igor Yu.

    2017-03-01

    In this work Ag nanoparticles (NPs) stable colloid solution were prepared by laser ablation of chemically pure silver rod in liquid monomer isodecyl acrylate (IDA). Sizes of obtained nanoparticles were determined by scanning electron microscope and vary from 27 to 120 nm. Nanocomposites films were prepared from obtained stable colloid solution of AgNPs by photocuring. For aliphatic polymer IDA long molecules cross-linking Diurethane dimethacrylate, 1,6-Hexandiol diacrylate and Tetra (ethylene glycol) diacrylate were used. Prepared nanomaterials exhibit strong third-order nonlinear optical responses, which was estimated by using z-scan technique. The third-order nonlinear optical coefficients of the studied nanocomposites were found to be up to Reχ(3)=1.31×10-5 (esu) and Imχ(3)=7.64×10-5 (esu).

  6. Multiphoton microscopy for the in-situ investigation of cellular processes and integrity in cryopreservation.

    PubMed

    Doerr, Daniel; Stark, Martin; Ehrhart, Friederike; Zimmermann, Heiko; Stracke, Frank

    2009-08-01

    In this study we demonstrate a new noninvasive imaging method to monitor freezing processes in biological samples and to investigate life in the frozen state. It combines a laser scanning microscope with a computer-controlled cryostage. Nearinfrared (NIR) femtosecond laser pulses evoke the fluorescence of endogenous fluorophores and fluorescent labels due to multiphoton absorption.The inherent optical nonlinearity of multiphoton absorption allows 3D fluorescence imaging for optical tomography of frozen biological material in-situ. As an example for functional imaging we use fluorescence lifetime imaging (FLIM) to create images with chemical and physical contrast.

  7. Characterization, Microbial Community Structure, and Pathogen Occurrence in Urban Faucet Biofilms in South China

    PubMed Central

    Lin, Huirong; Zhang, Shuting; Gong, Song; Zhang, Shenghua; Yu, Xin

    2015-01-01

    The composition and microbial community structure of the drinking water system biofilms were investigated using microstructure analysis and 454 pyrosequencing technique in Xiamen city, southeast of China. SEM (scanning electron microscope) results showed different features of biofilm morphology in different fields of PVC pipe. Extracellular matrix material and sparse populations of bacteria (mainly rod-shaped and coccoid) were observed. CLSM (confocal laser scanning microscope) revealed different distributions of attached cells, extracellular proteins, α-polysaccharides, and β-polysaccharides. The biofilms had complex bacterial compositions. Differences in bacteria diversity and composition from different tap materials and ages were observed. Proteobacteria was the common and predominant group in all biofilms samples. Some potential pathogens (Legionellales, Enterobacteriales, Chromatiales, and Pseudomonadales) and corrosive microorganisms were also found in the biofilms. This study provides the information of characterization and visualization of the drinking water biofilms matrix, as well as the microbial community structure and opportunistic pathogens occurrence. PMID:26273617

  8. Simultaneous photoacoustic and optically mediated ultrasound microscopy: an in vivo study

    PubMed Central

    Orlova, Anna; Shirmanova, Marina; Postnikova, Anna; Turchin, Ilya

    2015-01-01

    We propose the use of thermoelastic (TE) excitation of an ultrasonic (US) detector by backscattered laser radiation as a means of upgrading a single-modality photoacoustic (PA) microscope to dual-modality PA/US imaging at minimal cost. The upgraded scanning head of our dual-modality microscope consists of a fiber bundle with 14 output arms and a 32MHz polyvinylidene difluoride (PVDF) detector with a 34 MHz bandwidth (−6 dB level), 12.7 mm focal length, and a 0.25 numerical aperture. A single optical pulse delivered through the fiber bundle to the biotissue being investigated, in combination with a metalized surface on the PVDF detector allows us to obtain both PA and US A-scans. To demonstrate the in vivo capabilities of the proposed method we present the results of bimodal imaging of the brain of a newborn rat, a mouse tail and a mouse tumor. PMID:25780752

  9. Research of the Dispersity of the Functional Sericite/Methylphenyl- Silicone Resin

    PubMed Central

    Jiang, B.; Zhu, C. C.; Huang, Y. D.

    2015-01-01

    In order to improve the homogeneity and dispersity of the sericite in methylphenyl-silicone resin, the agglomerate state of the sericites was controlled effectively. The dispersive model of the sericite in methylphenyl-silicone resin was designed also. First, the modified sericite was prepared using hexadecyl trimethyl ammonium bromide as the intercalating agent. Then, functional sericite was incorporated into methylphenyl-silicone by terminal hydroxyl. The structure and dispersive performance of the hybrid polymers was charactered by analytical instruments. Scanning electron microscopy and Transmission electron microscope, Laser scanning confocal microscope and X-ray diffraction analysis showed that functional sericite was dispersed homogeneously in methylphenyl-silicone resin matrix. X-ray photoelectron spectroscopy analysis showed that the absorption peaks of the Si-OH band of methylphenyl-silicone resin were decreased and the Si-O-Si band was increased. This change evidently showed a significant role to enhance the reaction degree of the functional sericite in methylphenyl-silicone resin. PMID:26061002

  10. A novel system for water soluble protein encapsulation with high efficiency: "micelles enhanced" polyelectrolyte capsules.

    PubMed

    Li, Xiaodong; Li, Xiaohui; Zhang, Jianxiang; Zhao, Shifang; Shen, Jiacong

    2008-06-01

    Novel "micelles enhanced" polyelectrolyte (PE) capsules based on functional templates of hybrid calcium carbonate were fabricated. Evidences suggested that the structure of capsule wall was different from that of conventional PE capsules, and the wall permeability of these PE capsules changed significantly. Lysozyme, a positively charged protein in neutral solution, was studied as a model protein to be encapsulated into the "micelles enhanced" PE capsules. Confocal laser scanning microscope was used to observe the entrapping process in real time, while UV-Vis spectroscope and scanning force microscope measurements suggested the high efficiency of encapsulation. In addition, the fluorescence recovery after photobleaching technique was employed to determine the existence form of deposited molecules. Further studies showed even negatively charged water-soluble peptides or proteins can be encapsulated into these hybrid capsules by modulating the pH value in bulk solution under its isoelectronic point as well. Copyright 2007 Wiley Periodicals, Inc.

  11. Morphology variation, composition alteration and microstructure changes in ion-irradiated 1060 aluminum alloy

    NASA Astrophysics Data System (ADS)

    Wan, Hao; Si, Naichao; Wang, Quan; Zhao, Zhenjiang

    2018-02-01

    Morphology variation, composition alteration and microstructure changes in 1060 aluminum irradiated with 50 keV helium ions were characterized by field emission scanning electron microscopy (FESEM) equipped with x-ray elemental scanning, 3D measuring laser microscope and transmission electron microscope (TEM). The results show that, helium ions irradiation induced surface damage and Si-rich aggregates in the surfaces of irradiated samples. Increasing the dose of irradiation, more damages and Si-rich aggregates would be produced. Besides, defects such as dislocations, dislocation loops and dislocation walls were the primary defects in the ion implanted layer. The forming of surface damages were related with preferentially sputtering of Al component. While irradiation-enhanced diffusion and irradiation-induced segregation resulted in the aggregation of impurity atoms. And the aggregation ability of impurity atoms were discussed based on the atomic radius, displacement energy, lattice binding energy and surface binding energy.

  12. Characterization of a Laser Surface-Treated Martensitic Stainless Steel.

    PubMed

    Al-Sayed, S R; Hussein, A A; Nofal, A A; Hassab Elnaby, S I; Elgazzar, H

    2017-05-29

    Laser surface treatment was carried out on AISI 416 machinable martensitic stainless steel containing 0.225 wt.% sulfur. Nd:YAG laser with a 2.2-KW continuous wave was used. The aim was to compare the physical and chemical properties achieved by this type of selective surface treatment with those achieved by the conventional treatment. Laser power of different values (700 and 1000 W) with four corresponding different laser scanning speeds (0.5, 1, 2, and 3 m•min-1) was adopted to reach the optimum conditions for impact toughness, wear, and corrosion resistance for laser heat treated (LHT) samples. The 0 °C impact energy of LHT samples indicated higher values compared to the conventionally heat treated (CHT) samples. This was accompanied by the formation of a hard surface layer and a soft interior base metal. Microhardness was studied to determine the variation of hardness values with respect to the depth under the treated surface. The wear resistance at the surface was enhanced considerably. Microstructure examination was characterized using optical and scanning electron microscopes. The corrosion behavior of the LHT samples was also studied and its correlation with the microstructures was determined. The corrosion data was obtained in 3.5% NaCl solution at room temperature by means of a potentiodynamic polarization technique.

  13. Characterization of a Laser Surface-Treated Martensitic Stainless Steel

    PubMed Central

    Al-Sayed, S. R.; Hussein, A. A.; Nofal, A. A.; Hassab Elnaby, S. I.; Elgazzar, H.

    2017-01-01

    Laser surface treatment was carried out on AISI 416 machinable martensitic stainless steel containing 0.225 wt.% sulfur. Nd:YAG laser with a 2.2-KW continuous wave was used. The aim was to compare the physical and chemical properties achieved by this type of selective surface treatment with those achieved by the conventional treatment. Laser power of different values (700 and 1000 W) with four corresponding different laser scanning speeds (0.5, 1, 2, and 3 m·min−1) was adopted to reach the optimum conditions for impact toughness, wear, and corrosion resistance for laser heat treated (LHT) samples. The 0 °C impact energy of LHT samples indicated higher values compared to the conventionally heat treated (CHT) samples. This was accompanied by the formation of a hard surface layer and a soft interior base metal. Microhardness was studied to determine the variation of hardness values with respect to the depth under the treated surface. The wear resistance at the surface was enhanced considerably. Microstructure examination was characterized using optical and scanning electron microscopes. The corrosion behavior of the LHT samples was also studied and its correlation with the microstructures was determined. The corrosion data was obtained in 3.5% NaCl solution at room temperature by means of a potentiodynamic polarization technique. PMID:28772955

  14. Effects of laser polishing on surface microstructure and corrosion resistance of additive manufactured CoCr alloys

    NASA Astrophysics Data System (ADS)

    Wang, W. J.; Yung, K. C.; Choy, H. S.; Xiao, T. Y.; Cai, Z. X.

    2018-06-01

    Laser polishing of 3D printed metal components has drawn great interest in view of its potential applications in the dental implant industries. In this study, corrosion resistance, surface composition and crystalline structure of CoCr alloys were investigated. The corrosion resistance, micromorphology, composition, phase transformations and crystalline structures of samples were characterized using an electrochemical analyzer, scanning electron microscope (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and transmission electron microscope (TEM), respectively. The results indicate that high laser powers and low object distances within a certain range can facilitate the formation of complex oxide films, which exhibits high corrosion resistance. Further, object distances have a significant influence on cooling rates during the solidification of the melt pool in laser polishing, and fast cooling generates vast amounts of vacancies and defects, which result in the crystalline phase transformation from γ to ε. Consequently, the formed oxides play an important role in corrosion resistance on the outer layer, and inner layer with γ phase also helps keep the CoCr alloys in a stable structure with high resistant to corrosion. The two process parameters in laser polishing, laser power and object distances, are demonstrated as being important for controlling the surface microstructures and corrosion resistance of the additive manufactured CoCr alloy components.

  15. Nanosurgery of cells and chromosomes using near-infrared twelve-femtosecond laser pulses.

    PubMed

    Uchugonova, Aisada; Lessel, Matthias; Nietzsche, Sander; Zeitz, Christian; Jacobs, Karin; Lemke, Cornelius; König, Karsten

    2012-10-01

    ABSTRACT. Laser-assisted surgery based on multiphoton absorption of near-infrared laser light has great potential for high precision surgery at various depths within the cells and tissues. Clinical applications include refractive surgery (fs-LASIK). The non-contact laser method also supports contamination-free cell nanosurgery. In this paper we describe usage of an ultrashort femtosecond laser scanning microscope for sub-100 nm surgery of human cells and metaphase chromosomes. A mode-locked 85 MHz Ti:Sapphire laser with an M-shaped ultrabroad band spectrum (maxima: 770  nm/830  nm) and an in situ pulse duration at the target ranging from 12 fs up to 3 ps was employed. The effects of laser nanoprocessing in cells and chromosomes have been quantified by atomic force microscopy. These studies demonstrate the potential of extreme ultrashort femtosecond laser pulses at low mean milliwatt powers for sub-100 nm surgery of cells and cellular organelles.

  16. Doxorubicin-loaded Zein in situ gel for interstitial chemotherapy.

    PubMed

    Cao, Xiaoying; Geng, Jianning; Su, Suwen; Zhang, Linan; Xu, Qian; Zhang, Li; Xie, Yinghua; Wu, Shaomei; Sun, Yongjun; Gao, Zibin

    2012-01-01

    A novel drug delivery system of doxorubicin (DOX)-loaded Zein in situ gel for interstitial chemotherapy was investigated in this study. The possible mechanisms of drug release were described according to morphological analysis by optical microscopy and scanning electronic microscope (SEM). In vitro and in vivo anti-tumor activity studies showed that DOX-loaded Zein in situ gel was superior to DOX solution. Local pharmacokinetics in tumor tissue was studied by quantitative analysis with confocal laser scanning microscopy (CLSM) combined with microdialysis technology. A pharmacokinetics mathematical model of DOX-loaded Zein in situ gel in tumors was then built.

  17. Safety of laser use under the dental microscope.

    PubMed

    Saegusa, Hidetoshi; Watanabe, Satoshi; Anjo, Tomoo; Ebihara, Arata; Suda, Hideaki

    2010-04-01

    The aim of this study was to investigate the safety of laser use under the dental microscope. Nd:YAG, Er:YAG and diode lasers were used. The end of the tips was positioned at a distance of 5 cm from the objective lens of a dental microscope. Each eye protector was made into a flat disc, which was fixed on the lens of the microscope. The filters were placed in front of the objective lens or behind the eye lens. Transmitted energy through the microscope with or without the filters was measured. No transmitted laser energy was detected when using matched eye protectors. Mismatched eye protectors were not effective for shutting out laser energy, especially for Nd:YAG and diode lasers. None or very little laser energy was detected through the microscope even without any laser filter. Matched filters shut out all laser energy irrespective of their positions.

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

    NASA Astrophysics Data System (ADS)

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

    2017-02-01

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

  19. Influence of operating microscope in the sealing of cervical perforations.

    PubMed

    Schmidt, Bruna Schwingel; Zaccara, Ivana Maria; Reis Só, Marcus Vinícius; Kuga, Milton Carlos; Palma-Dibb, Regina Guenka; Kopper, Patrícia Maria Poli

    2016-01-01

    Accidental root canal perforations are among the main complications of endodontic treatment. This study evaluated the influence of operating microscope (OM) in the marginal adaptation of mineral trioxide aggregate (MTA) (Angelus(®)) and glass ionomer (Vitremer) inserted into cervical perforations. Perforations were made in the cervical third of the buccal wall of the root canal in mandibular incisors. Next, the teeth were divided into four groups (N = 10): MG - MTA without OM; VG - Vitremer without OM; MOMG - MTA with OM; VOMG - Vitremer with OM. The perforations were sealed according to the group and the teeth were prepared for analysis by confocal laser scanning microscope. Images of perforation region (1,024×) were made and the gap presented by the materials was measured using the Image J program. LEXT OLS4100 three dimensional (3D) measuring laser microscope measured the volumetric misfit. Data of gap were analyzed by Kruskal-Wallis and Dunn's tests. Analysis of variance (ANOVA) and Tukey's tests compared the volumetric misfits. The results showed lower volume and gap in the interface dentin/material in VOMG compared to the other groups (P < 0.05). The use of OM improved the quality of cervical perforations sealed with Vitremer, being indicated in clinical situations of iatrogenic cervical perforations.

  20. Collection of trace evidence of explosive residues from the skin in a death due to a disguised letter bomb. The synergy between confocal laser scanning microscope and inductively coupled plasma atomic emission spectrometer analyses.

    PubMed

    Turillazzi, Emanuela; Monaci, Fabrizio; Neri, Margherita; Pomara, Cristoforo; Riezzo, Irene; Baroni, Davide; Fineschi, Vittorio

    2010-04-15

    In most deaths caused by explosive, the victim's body becomes a depot for fragments of explosive materials, so contributing to the collection of trace evidence which may provide clues about the specific type of device used with explosion. Improvised explosive devices are used which contain "homemade" explosives rather than high explosives because of the relative ease with which such components can be procured. Many methods such as chromatography-mass spectrometry, scanning electron microscopy, stereomicroscopy, capillary electrophoresis are available for use in the identification of explosive residues on objects and bomb fragments. Identification and reconstruction of the distribution of explosive residues on the decedent's body may give additional hints in assessing the position of the victim in relation to the device. Traditionally these residues are retrieved by swabbing the body and clothing during the early phase, at autopsy. Gas chromatography-mass spectrometry and other analytical methods may be used to analyze the material swabbed from the victim body. The histological examination of explosive residues on skin samples collected during the autopsy may reveal significant details. The information about type, quantity and particularly about anatomical distribution of explosive residues obtained utilizing confocal laser scanning microscope (CLSM) together with inductively coupled plasma atomic emission spectrometer (ICP-AES), may provide very significant evidence in the clarification and reconstruction of the explosive-related events. Copyright 2009 Elsevier Ireland Ltd. All rights reserved.

  1. Microscope self-calibration based on micro laser line imaging and soft computing algorithms

    NASA Astrophysics Data System (ADS)

    Apolinar Muñoz Rodríguez, J.

    2018-06-01

    A technique to perform microscope self-calibration via micro laser line and soft computing algorithms is presented. In this technique, the microscope vision parameters are computed by means of soft computing algorithms based on laser line projection. To implement the self-calibration, a microscope vision system is constructed by means of a CCD camera and a 38 μm laser line. From this arrangement, the microscope vision parameters are represented via Bezier approximation networks, which are accomplished through the laser line position. In this procedure, a genetic algorithm determines the microscope vision parameters by means of laser line imaging. Also, the approximation networks compute the three-dimensional vision by means of the laser line position. Additionally, the soft computing algorithms re-calibrate the vision parameters when the microscope vision system is modified during the vision task. The proposed self-calibration improves accuracy of the traditional microscope calibration, which is accomplished via external references to the microscope system. The capability of the self-calibration based on soft computing algorithms is determined by means of the calibration accuracy and the micro-scale measurement error. This contribution is corroborated by an evaluation based on the accuracy of the traditional microscope calibration.

  2. Nanosecond laser-induced ablation and laser-induced shockwave structuring of polymer foils down to sub-μm patterns

    NASA Astrophysics Data System (ADS)

    Lorenz, P.; Bayer, L.; Ehrhardt, M.; Zimmer, K.; Engisch, L.

    2015-03-01

    Micro- and nanostructures exhibit a growing commercial interest where a fast, cost-effective, and large-area production is attainable. Laser methods have a great potential for the easy fabrication of surface structures into flexible polymer foils like polyimide (PI). In this study two different concepts for the structuring of polymer foils using a KrF excimer laser were tested and compared: the laser-induced ablation and the laser-induced shock wave structuring. The direct front side laser irradiation of these polymers allows the fabrication of different surface structures. For example: The low laser fluence treatment of PI results in nano-sized cone structures where the cone density can be controlled by the laser parameters. This allows inter alia the laser fabrication of microscopic QR code and high-resolution grey-tone images. Furthermore, the laser treatment of the front side of the polymer foil allows the rear side structuring due to a laserinduced shock wave. The resultant surface structures were analysed by optical and scanning electron microscopy (SEM) as well as white light interferometry (WLI).

  3. Femtosecond laser etching of dental enamel for bracket bonding.

    PubMed

    Kabas, Ayse Sena; Ersoy, Tansu; Gülsoy, Murat; Akturk, Selcuk

    2013-09-01

    The aim is to investigate femtosecond laser ablation as an alternative method for enamel etching used before bonding orthodontic brackets. A focused laser beam is scanned over enamel within the area of bonding in a saw tooth pattern with a varying number of lines. After patterning, ceramic brackets are bonded and bonding quality of the proposed technique is measured by a universal testing machine. The results are compared to the conventional acid etching method. Results show that bonding strength is a function of laser average power and the density of the ablated lines. Intrapulpal temperature changes are also recorded and observed minimal effects are observed. Enamel surface of the samples is investigated microscopically and no signs of damage or cracking are observed. In conclusion, femtosecond laser exposure on enamel surface yields controllable patterns that provide efficient bonding strength with less removal of dental tissue than conventional acid-etching technique.

  4. Effect of Laser Power on Metallurgical, Mechanical and Tribological Characteristics of Hardfaced Surfaces of Nickel-Based Alloy

    NASA Astrophysics Data System (ADS)

    Gnanasekaran, S.; Padmanaban, G.; Balasubramanian, V.

    2017-12-01

    In this present work, nickel based alloy was deposited on 316 LN austenitic stainless steel (ASS) by a laser hardfacing technique to investigate the influence of laser power on macrostructure, microstructure, microhardness, dilution and wear characteristics. The laser power varied from 1.1 to 1.9 kW. The phase constitution, microstructure and microhardness were examined by optical microscope, scanning electron microscopy, energy dispersion spectroscopy and Vickers microhardness tester. The wear characteristics of the hardfaced surfaces and substrate were evaluated at room temperature (RT) under dry sliding wear condition (pin-on-disc). The outcome demonstrates that as the laser power increases, dilution increases and hardness of the deposit decreases. This is because excess heat melts more volume of substrate material and increases the dilution; subsequently it decreases the hardness of the deposit. The microstructure of the deposit is characterized by Ni-rich carbide, boride and silicide.

  5. Bond strengths evaluation of laser ceramic bracket debonding

    NASA Astrophysics Data System (ADS)

    Dostalová, T.; Jelinková, H.; Šulc, J.; Němec, M.; Fibrich, M.; Jelínek, M.; Michalík, P.; Bučková, M.

    2012-09-01

    Ceramic brackets often used for an orthodontic treatment can lead to problems such as enamel tear outs because of their low fracture resistance and high bond strengths. Therefore the aim of our study was to investigate the positive laser radiation effect on bracket debonding. Moreover, the influence of the enamel shape surface under the bracket and laser radiation power on the debonding strength was investigated. The source of the radiation was the longitudinally diode-pumped Tm:YAP laser operating at 1997 nm. To eliminate the tooth surface roughness the flat enamel surface was prepared artificially and the bracket was bonded on it. The debonding was accomplished by Tm:YAP laser radiation with different the power value while recording the temperature rise in the pulp. To simulate the debonding process in vivo the actual bond strength was measured by the digital force gauge. The results were analyzed by scanning electron microscope.

  6. Identification of endogenous fluorophores in the photoreceptors using autofluorescence spectroscopy

    NASA Astrophysics Data System (ADS)

    Zhao, Lingling; Qu, Junle; Niu, Hanben

    2007-11-01

    In this paper, we present our investigation on the identification of endogenous fluorophores in photoreceptors using autofluorescence spectroscopy, which is performed with an inverted laser scanning confocal microscope equipped with an Argon ion laser and a GreNe laser. In our experiments, individual cones and rods are clearly resolved even in freshly prepared retina samples, without slicing or labeling. The experiment results show that autofluorescence spectrum of the photoreceptors has three peaks approximately at 525nm, 585nm and 665nm. Furthermore, the brightest autofluorescence originates from the photoreceptor outer segments. We can, therefore, come to a conclusion that the peaks at 525nm, 585nm are corresponding to FAD and A2-PE, respectively, which are distributed in the photoreceptor outer segments.

  7. Effect of pulse temporal shape on optical trapping and impulse transfer using ultrashort pulsed lasers.

    PubMed

    Shane, Janelle C; Mazilu, Michael; Lee, Woei Ming; Dholakia, Kishan

    2010-03-29

    We investigate the effects of pulse duration on optical trapping with high repetition rate ultrashort pulsed lasers, through Lorentz-Mie theory, numerical simulation, and experiment. Optical trapping experiments use a 12 femtosecond duration infrared pulsed laser, with the trapping microscope's temporal dispersive effects measured and corrected using the Multiphoton Intrapulse Interference Phase Scan method. We apply pulse shaping to reproducibly stretch pulse duration by 1.5 orders of magnitude and find no material-independent effects of pulse temporal profile on optical trapping of 780nm silica particles, in agreement with our theory and simulation. Using pulse shaping, we control two-photon fluorescence in trapped fluorescent particles, opening the door to other coherent control applications with trapped particles.

  8. Nonlinear laser scanning microscopy of human vocal folds.

    PubMed

    Miri, Amir K; Tripathy, Umakanta; Mongeau, Luc; Wiseman, Paul W

    2012-02-01

    The purpose of this work was to apply nonlinear laser scanning microscopy (NLSM) for visualizing the morphology of extracellular matrix proteins within human vocal folds. This technique may potentially assist clinicians in making rapid diagnoses of vocal fold tissue disease or damage. Microstructural characterization based on NLSM provides valuable information for better understanding molecular mechanisms and tissue structure. Experimental, ex vivo human vocal fold. A custom-built multimodal nonlinear laser scanning microscope was used to scan fibrillar proteins in three 4% formaldehyde-fixed cadaveric samples. Collagen and elastin, key extracellular matrix proteins in the vocal fold lamina propria, were imaged by two nonlinear microscopy modalities: second harmonic generation (SHG) and two-photon fluorescence (TPF), respectively. An experimental protocol was introduced to characterize the geometrical properties of the imaged fibrous proteins. NLSM revealed the biomorphology of the human vocal fold fibrous proteins. No photobleaching was observed for the incident laser power of ∼60 mW before the excitation objective. Types I and III fibrillar collagen were imaged without label in the tissue by intrinsic SHG. Imaging while rotating the incident laser light-polarization direction confirmed a helical shape for the collagen fibers. The amplitude, periodicity, and overall orientation were then computed for the helically distributed collagen network. The elastin network was simultaneously imaged via TPF and found to have a basket-like structure. In some regions, particularly close to the epithelium, colocalization of both extracellular matrix components were observed. A benchmark study is presented for quantitative real-time, ex vivo, NLSM imaging of the extracellular macromolecules in human vocal fold lamina propria. The results are promising for clinical applications. Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.

  9. Imaging Single Cells in the Living Retina

    PubMed Central

    Williams, David R.

    2011-01-01

    A quarter century ago, we were limited to a macroscopic view of the retina inside the living eye. Since then, new imaging technologies, including confocal scanning laser ophthalmoscopy, optical coherence tomography, and adaptive optics fundus imaging, transformed the eye into a microscope in which individual cells can now be resolved noninvasively. These technologies have enabled a wide range of studies of the retina that were previously impossible. PMID:21596053

  10. Fluorescence Microscopy Gets Faster and Clearer: Roles of Photochemistry and Selective Illumination

    PubMed Central

    Wolenski, Joseph S.; Julich, Doerthe

    2014-01-01

    Significant advances in fluorescence microscopy tend be a balance between two competing qualities wherein improvements in resolution and low light detection are typically accompanied by losses in acquisition rate and signal-to-noise, respectively. These trade-offs are becoming less of a barrier to biomedical research as recent advances in optoelectronic microscopy and developments in fluorophore chemistry have enabled scientists to see beyond the diffraction barrier, image deeper into live specimens, and acquire images at unprecedented speed. Selective plane illumination microscopy has provided significant gains in the spatial and temporal acquisition of fluorescence specimens several mm in thickness. With commercial systems now available, this method promises to expand on recent advances in 2-photon deep-tissue imaging with improved speed and reduced photobleaching compared to laser scanning confocal microscopy. Superresolution microscopes are also available in several modalities and can be coupled with selective plane illumination techniques. The combination of methods to increase resolution, acquisition speed, and depth of collection are now being married to common microscope systems, enabling scientists to make significant advances in live cell and in situ imaging in real time. We show that light sheet microscopy provides significant advantages for imaging live zebrafish embryos compared to laser scanning confocal microscopy. PMID:24600334

  11. A Generalization of Theory for Two-Dimensional Fluorescence Recovery after Photobleaching Applicable to Confocal Laser Scanning Microscopes

    PubMed Central

    Kang, Minchul; Day, Charles A.; Drake, Kimberly; Kenworthy, Anne K.; DiBenedetto, Emmanuele

    2009-01-01

    Abstract Fluorescence recovery after photobleaching (FRAP) using confocal laser scanning microscopes (confocal FRAP) has become a valuable technique for studying the diffusion of biomolecules in cells. However, two-dimensional confocal FRAP sometimes yields results that vary with experimental setups, such as different bleaching protocols and bleaching spot sizes. In addition, when confocal FRAP is used to measure diffusion coefficients (D) for fast diffusing molecules, it often yields D-values that are one or two orders-of-magnitude smaller than that predicted theoretically or measured by alternative methods such as fluorescence correlation spectroscopy. Recently, it was demonstrated that this underestimation of D can be corrected by taking diffusion during photobleaching into consideration. However, there is currently no consensus on confocal FRAP theory, and no efforts have been made to unify theories on conventional and confocal FRAP. To this end, we generalized conventional FRAP theory to incorporate diffusion during photobleaching so that analysis by conventional FRAP theory for a circular region of interest is easily applicable to confocal FRAP. Finally, we demonstrate the accuracy of these new (to our knowledge) formulae by measuring D for soluble enhanced green fluorescent protein in aqueous glycerol solution and in the cytoplasm and nucleus of COS7 cells. PMID:19720039

  12. Effect of engraving speeds of CO₂ laser irradiation on In-Ceram Alumina roughness: a pilot study.

    PubMed

    Ersu, Bahadır; Ersoy, Orkun; Yuzugullu, Bulem; Canay, Senay

    2015-05-01

    The aim of the study was to determine the effect of CO₂ laser on surface roughness of In-Ceram-Alumina-ceramic. Four aluminum-oxide ceramic disc specimens were prepared of In-Ceram Alumina. Discs received CO₂ laser irradiation with different engraving speeds (100, 400, 600 and 800 mm/min) as a surface treatment. The roughness of the surfaces was measured on digital elevation models reconstructed from stereoscopic images acquired by scanning-electron-microscope. Surface roughness data were analyzed with One-Way-Analysis-of-Variance at a significance level of p<0.05. There was no significant difference between the roughness values (p=0.82). Due to higher laser durations, partial melting signs were observed on the surfaces. Tearing, smearing and swelling occurred on melted surfaces. Swelling accompanying melting increased the surface roughness, while laser power was fixed and different laser engraving speeds were applied. Although different laser irradiation speeds did not affect the roughness of ceramic surfaces, swelling was observed which led to changes on surfaces.

  13. Effect of photon-initiated photoacoustic streaming, passive ultrasonic, and sonic irrigation techniques on dentinal tubule penetration of irrigation solution: a confocal microscopic study.

    PubMed

    Akcay, Merve; Arslan, Hakan; Mese, Merve; Durmus, Nazlı; Capar, Ismail Davut

    2017-09-01

    The aim of this in vitro study was to evaluate the efficacy of different irrigation techniques including laser-activated irrigation using an erbium:yttrium-aluminum-garnet (Er:YAG) laser with a novel tip design (photon-induced photoacoustic streaming (PIPS)), Er:YAG laser with Preciso tip, sonic activation, and passive ultrasonic activation on the final irrigation solution penetration into dentinal tubules by using a laser scanning confocal microscope. In this study, 65 extracted single-rooted human mandibular premolars were instrumented up to size 40 and randomly divided into 5 groups (n = 13) based on the activation technique of the final irrigation solution as follows: conventional irrigation (control group), sonic activation, passive ultrasonic activation, Er:YAG-PIPS tip activation, and Er:YAG-Preciso tip activation. In each group, 5 mL of 5% NaOCl labeled with fluorescent dye was used during the activation as the final irrigation solution. Specimens were sectioned at 2.5 and 8 mm from the apex and then examined under a confocal microscope to calculate the dentinal tubule penetration area. Data were analyzed using two-way analysis of variance (ANOVA) and Tukey's post hoc tests (P = 0.05). Both Er:YAG laser (Preciso/PIPS) activations exhibited a significantly higher penetration area than the other groups (P < 0.05). Additionally, passive ultrasonic activation had significantly higher penetration than the sonic activation group and the control group. Statistically significant differences were also found between each root canal third (coronal > middle > apical) (P < 0.001). The results from the present study support the use of Er:YAG laser activation (Preciso/PIPS) to improve the effectiveness of the final irrigation procedure by increasing the irrigant penetration area into the dentinal tubules. The activation of the irrigant and the creation of the streaming with the Er:YAG laser have a positive effect on the irrigant penetration.

  14. Nondestructive monitoring damage in composites using scanning laser acoustic microscopy

    NASA Technical Reports Server (NTRS)

    Wey, A. C.; Kessler, L. W.; Dos Reis, H. L. M.

    1992-01-01

    Several Nicalon fiber reinforced LAS (lithium alumino-silicate) glass matrix composites were tested to study the relation between the residual strength and the different amounts of damage. The samples were fatigued by four-point cyclic loading at a 5 Hz rate at 500 C for a different number of cycles. 10 MHz scanning laser acoustic microscope (SLAM) images were taken to monitor damage on the samples. Our SLAM results indicate that there were defects already existing throughout the sample before fatigue, and the resultant damage pattern from fatigue could be related to the initial defect distribution in the sample. Finally, the fatigued samples were fractured and the residual strength data could not be explained by the cyclic fatigue alone. Rather, the damage patterns evident in the SLAM images were needed to explain the scatter in the data. The results show that SLAM is useful in nondestructively monitoring damage and estimating residual strength of fatigued ceramic composites.

  15. Enhanced Thermo-Optical Switching of Paraffin-Wax Composite Spots under Laser Heating

    PubMed Central

    Said, Asmaa; Salah, Abeer; Abdel Fattah, Gamal

    2017-01-01

    Thermo-optical switches are of particular significance in communications networks where increasingly high switching speeds are required. Phase change materials (PCMs), in particular those based on paraffin wax, provide wealth of exciting applications with unusual thermally-induced switching properties, only limited by paraffin’s rather low thermal conductivity. In this paper, the use of different carbon fillers as thermal conductivity enhancers for paraffin has been investigated, and a novel structure based on spot of paraffin wax as a thermo-optic switch is presented. Thermo-optical switching parameters are enhanced with the addition of graphite and graphene, due to the extreme thermal conductivity of the carbon fillers. Differential Scanning Calorimetry (DSC) and Scanning electron microscope (SEM) are performed on paraffin wax composites, and specific heat capacities are calculated based on DSC measurements. Thermo-optical switching based on transmission is measured as a function of the host concentration under conventional electric heating and laser heating of paraffin-carbon fillers composites. Further enhancements in thermo-optical switching parameters are studied under Nd:YAG laser heating. This novel structure can be used in future networks with huge bandwidth requirements and electric noise free remote aerial laser switching applications. PMID:28772884

  16. Enhanced Thermo-Optical Switching of Paraffin-Wax Composite Spots under Laser Heating.

    PubMed

    Said, Asmaa; Salah, Abeer; Fattah, Gamal Abdel

    2017-05-12

    Thermo-optical switches are of particular significance in communications networks where increasingly high switching speeds are required. Phase change materials (PCMs), in particular those based on paraffin wax, provide wealth of exciting applications with unusual thermally-induced switching properties, only limited by paraffin's rather low thermal conductivity. In this paper, the use of different carbon fillers as thermal conductivity enhancers for paraffin has been investigated, and a novel structure based on spot of paraffin wax as a thermo-optic switch is presented. Thermo-optical switching parameters are enhanced with the addition of graphite and graphene, due to the extreme thermal conductivity of the carbon fillers. Differential Scanning Calorimetry (DSC) and Scanning electron microscope (SEM) are performed on paraffin wax composites, and specific heat capacities are calculated based on DSC measurements. Thermo-optical switching based on transmission is measured as a function of the host concentration under conventional electric heating and laser heating of paraffin-carbon fillers composites. Further enhancements in thermo-optical switching parameters are studied under Nd:YAG laser heating. This novel structure can be used in future networks with huge bandwidth requirements and electric noise free remote aerial laser switching applications.

  17. Transmissive liquid-crystal device correcting primary coma aberration and astigmatism in laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Tanabe, Ayano; Hibi, Terumasa; Ipponjima, Sari; Matsumoto, Kenji; Yokoyama, Masafumi; Kurihara, Makoto; Hashimoto, Nobuyuki; Nemoto, Tomomi

    2016-03-01

    Laser scanning microscopy allows 3D cross-sectional imaging inside biospecimens. However, certain aberrations produced can degrade the quality of the resulting images. We previously reported a transmissive liquid-crystal device that could compensate for the predominant spherical aberrations during the observations, particularly in deep regions of the samples. The device, inserted between the objective lens and the microscope revolver, improved the image quality of fixed-mouse-brain slices that were observed using two-photon excitation laser scanning microscopy, which was originally degraded by spherical aberration. In this study, we developed a transmissive device that corrects primary coma aberration and astigmatism, motivated by the fact that these asymmetric aberrations can also often considerably deteriorate image quality, even near the sample surface. The device's performance was evaluated by observing fluorescent beads using single-photon excitation laser scanning microscopy. The fluorescence intensity in the image of the bead under a cover slip tilted in the y-direction was increased by 1.5 times after correction by the device. Furthermore, the y- and z-widths of the imaged bead were reduced to 66% and 65%, respectively. On the other hand, for the imaged bead sucked into a glass capillary in the longitudinal x-direction, correction with the device increased the fluorescence intensity by 2.2 times compared to that of the aberrated image. In addition, the x-, y-, and z-widths of the bead image were reduced to 75%, 53%, and 40%, respectively. Our device successfully corrected several asymmetric aberrations to improve the fluorescent signal and spatial resolution, and might be useful for observing various biospecimens.

  18. Fabrication of 2D and 3D photonic structures using laser lithography

    NASA Astrophysics Data System (ADS)

    Gaso, P.; Jandura, D.; Pudis, D.

    2016-12-01

    In this paper we demonstrate possibilities of three-dimensional (3D) printing technology based on two photon polymerization. We used three-dimensional dip-in direct-laser-writing (DLW) optical lithography to fabricate 2D and 3D optical structures for optoelectronics and for optical sensing applications. DLW lithography allows us use a non conventional way how to couple light into the waveguide structure. We prepared ring resonator and we investigated its transmission spectral characteristic. We present 3D inverse opal structure from its design to printing and scanning electron microscope (SEM) imaging. Finally, SEM images of some prepared photonic crystal structures were performed.

  19. Low-power noncontact photoacoustic microscope for bioimaging applications

    NASA Astrophysics Data System (ADS)

    Sathiyamoorthy, Krishnan; Strohm, Eric M.; Kolios, Michael C.

    2017-04-01

    An inexpensive noncontact photoacoustic (PA) imaging system using a low-power continuous wave laser and a kilohertz-range microphone has been developed. The system operates in both optical and PA imaging modes and is designed to be compatible with conventional optical microscopes. Aqueous coupling fluids are not required for the detection of the PA signals; air is used as the coupling medium. The main component of the PA system is a custom designed PA imaging sensor that consists of an air-filled sample chamber and a resonator chamber that isolates a standard kilohertz frequency microphone from the input laser. A sample to be examined is placed on the glass substrate inside the chamber. A laser focused to a small spot by a 40× objective onto the substrate enables generation of PA signals from the sample. Raster scanning the laser over the sample with micrometer-sized steps enables high-resolution PA images to be generated. A lateral resolution of 1.37 μm was achieved in this proof of concept study, which can be further improved using a higher numerical aperture objective. The application of the system was investigated on a red blood cell, with a noise-equivalent detection sensitivity of 43,887 hemoglobin molecules (72.88×10-21 mol or 72.88 zeptomol). The minimum pressure detectable limit of the system was 19.1 μPa. This inexpensive, compact noncontact PA sensor is easily integrated with existing commercial optical microscopes, enabling optical and PA imaging of the same sample. Applications include forensic measurements, blood coagulation tests, and monitoring the penetration of drugs into human membrane.

  20. Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity

    PubMed Central

    Boudreau, Colton; Wee, Tse-Luen (Erika); Duh, Yan-Rung (Silvia); Couto, Melissa P.; Ardakani, Kimya H.; Brown, Claire M.

    2016-01-01

    It is important to determine the most effective method of delivering light onto a specimen for minimal light induced damage. Assays are presented to measure photo-bleaching of fluorophores and photo-toxicity to living cells under different illumination conditions. Turning the light off during part of the experimental time reduced photo-bleaching in a manner proportional to the time of light exposure. The rate of photo-bleaching of EGFP was reduced by 9-fold with light pulsing on the micro-second scale. Similarly, in living cells, rapid line scanning resulted in reduced cell stress as measured by mitochondrial potential, rapid cell protrusion and reduced cell retraction. This was achieved on a commercial confocal laser scanning microscope, without any compromise in image quality, by using rapid laser scan settings and line averaging. Therefore this technique can be implemented broadly without any software or hardware upgrades. Researchers can use the rapid line scanning option to immediately improve image quality on fixed samples, reduce photo-bleaching for large high resolution 3D datasets and improve cell health in live cell experiments. The assays developed here can be applied to other microscopy platforms to measure and optimize light delivery for minimal sample damage and photo-toxicity. PMID:27485088

  1. Excitation Light Dose Engineering to Reduce Photo-bleaching and Photo-toxicity.

    PubMed

    Boudreau, Colton; Wee, Tse-Luen Erika; Duh, Yan-Rung Silvia; Couto, Melissa P; Ardakani, Kimya H; Brown, Claire M

    2016-08-03

    It is important to determine the most effective method of delivering light onto a specimen for minimal light induced damage. Assays are presented to measure photo-bleaching of fluorophores and photo-toxicity to living cells under different illumination conditions. Turning the light off during part of the experimental time reduced photo-bleaching in a manner proportional to the time of light exposure. The rate of photo-bleaching of EGFP was reduced by 9-fold with light pulsing on the micro-second scale. Similarly, in living cells, rapid line scanning resulted in reduced cell stress as measured by mitochondrial potential, rapid cell protrusion and reduced cell retraction. This was achieved on a commercial confocal laser scanning microscope, without any compromise in image quality, by using rapid laser scan settings and line averaging. Therefore this technique can be implemented broadly without any software or hardware upgrades. Researchers can use the rapid line scanning option to immediately improve image quality on fixed samples, reduce photo-bleaching for large high resolution 3D datasets and improve cell health in live cell experiments. The assays developed here can be applied to other microscopy platforms to measure and optimize light delivery for minimal sample damage and photo-toxicity.

  2. Comparative study viruses with computer-aided phase microscope AIRYSCAN

    NASA Astrophysics Data System (ADS)

    Tychinsky, Vladimir P.; Koufal, Georgy E.; Perevedentseva, Elena V.; Vyshenskaia, Tatiana V.

    1996-12-01

    Traditionally viruses are studied with scanning electron microscopy (SEM) after complicated procedure of sample preparation without the possibility to study it under natural conditions. We obtained images of viruses (Vaccinia virus, Rotavirus) and rickettsias (Rickettsia provazekii, Coxiella burnetti) in native state with computer-aided phase microscope airyscan -- the interference microscope of Linnik layout with phase modulation of the reference wave with dissector image tube as coordinate-sensitive photodetector and computer processing of phase image. A light source was the He-Ne laser. The main result is coincidence of dimensions and shape of phase images with available information concerning their morphology obtained with SEM and other methods. The fine structure of surface and nuclei is observed. This method may be applied for virus recognition and express identification, investigation of virus structure and the analysis of cell-virus interaction.

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

    PubMed

    Bruns, T; Schickinger, S; Schneckenburger, H

    2015-10-01

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

  4. HelioScan: a software framework for controlling in vivo microscopy setups with high hardware flexibility, functional diversity and extendibility.

    PubMed

    Langer, Dominik; van 't Hoff, Marcel; Keller, Andreas J; Nagaraja, Chetan; Pfäffli, Oliver A; Göldi, Maurice; Kasper, Hansjörg; Helmchen, Fritjof

    2013-04-30

    Intravital microscopy such as in vivo imaging of brain dynamics is often performed with custom-built microscope setups controlled by custom-written software to meet specific requirements. Continuous technological advancement in the field has created a need for new control software that is flexible enough to support the biological researcher with innovative imaging techniques and provide the developer with a solid platform for quickly and easily implementing new extensions. Here, we introduce HelioScan, a software package written in LabVIEW, as a platform serving this dual role. HelioScan is designed as a collection of components that can be flexibly assembled into microscope control software tailored to the particular hardware and functionality requirements. Moreover, HelioScan provides a software framework, within which new functionality can be implemented in a quick and structured manner. A specific HelioScan application assembles at run-time from individual software components, based on user-definable configuration files. Due to its component-based architecture, HelioScan can exploit synergies of multiple developers working in parallel on different components in a community effort. We exemplify the capabilities and versatility of HelioScan by demonstrating several in vivo brain imaging modes, including camera-based intrinsic optical signal imaging for functional mapping of cortical areas, standard two-photon laser-scanning microscopy using galvanometric mirrors, and high-speed in vivo two-photon calcium imaging using either acousto-optic deflectors or a resonant scanner. We recommend HelioScan as a convenient software framework for the in vivo imaging community. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Capillary array scanner for time-resolved detection and identification of fluorescently labelled DNA fragments.

    PubMed

    Neumann, M; Herten, D P; Dietrich, A; Wolfrum, J; Sauer, M

    2000-02-25

    The first capillary array scanner for time-resolved fluorescence detection in parallel capillary electrophoresis based on semiconductor technology is described. The system consists essentially of a confocal fluorescence microscope and a x,y-microscope scanning stage. Fluorescence of the labelled probe molecules was excited using a short-pulse diode laser emitting at 640 nm with a repetition rate of 50 MHz. Using a single filter system the fluorescence decays of different labels were detected by an avalanche photodiode in combination with a PC plug-in card for time-correlated single-photon counting (TCSPC). The time-resolved fluorescence signals were analyzed and identified by a maximum likelihood estimator (MLE). The x,y-microscope scanning stage allows for discontinuous, bidirectional scanning of up to 16 capillaries in an array, resulting in longer fluorescence collection times per capillary compared to scanners working in a continuous mode. Synchronization of the alignment and measurement process were developed to allow for data acquisition without overhead. Detection limits in the subzeptomol range for different dye molecules separated in parallel capillaries have been achieved. In addition, we report on parallel time-resolved detection and separation of more than 400 bases of single base extension DNA fragments in capillary array electrophoresis. Using only semiconductor technology the presented technique represents a low-cost alternative for high throughput DNA sequencing in parallel capillaries.

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

    PubMed

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

    1998-11-01

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

  7. Laser texturing of Hastelloy C276 alloy surface for improved hydrophobicity and friction coefficient

    NASA Astrophysics Data System (ADS)

    Yilbas, B. S.; Ali, H.

    2016-03-01

    Laser treatment of Hastelloy C276 alloy is carried out under the high pressure nitrogen assisting gas environment. Morphological and metallurgical changes in the laser treated layer are examined using the analytical tools including, scanning electron and atomic force microscopes, X-ray diffraction, energy dispersive spectroscopy, and Fourier transform infrared spectroscopy. Microhardness is measured and the residual stress formed in the laser treated surface is determined from the X-ray data. The hydrophibicity of the laser treated surface is assessed using the sessile drop method. Friction coefficient of the laser treated layer is obtained incorporating the micro-tribometer. It is found that closely spaced laser canning tracks create a self-annealing effect in the laser treated layer and lowers the thermal stress levels through modifying the cooling rates at the surface. A dense structure, consisting of fine size grains, enhances the microhardness of the surface. The residual stress formed at the surface is compressive and it is in the order of -800 MPa. Laser treatment improves the surface hydrophobicity significantly because of the formation of surface texture composing of micro/nano-pillars.

  8. Femtosecond-laser assisted cell reprogramming

    NASA Astrophysics Data System (ADS)

    Breunig, Hans Georg; Uchugonova, Aisada; Batista, Ana; König, Karsten

    2017-02-01

    Femtosecond-laser pulses can assist to transfect cells by creating transient holes in the cell membrane, thus making them temporarily permeable for extraneous genetic material. This procedure offers the advantage of being completely "virus free" since no viruses are used for the delivery and integration of gene factors into the host genome and, thereby, avoiding serious side effects which so far prevent clinical application. Unfortunately, focusing of the laser radiation onto individual cell membranes is quite elaborate and time consuming. Regarding these obstacles, we briefly review two optical setups for fast, efficient and high throughput laser-assisted cell transfection based on femtosecond laser pulse excitation. The first setup aims at assisting the transfection of adherent cells. It comprises of a modified laser-scanning microscope with beamshaping optics as well as home-made software to automate the detection, targeting and laser-irradiation process. The second setup aims at laser-assisted transfection of non-adherent cells in suspension which move in a continuous flow through the laser focus region. The setup allows to address a large number of cells, however, with much lower transfection efficiency than the individual-cell targeting approach.

  9. Effects of laser-aided circumferential supracrestal fiberotomy on root surfaces.

    PubMed

    Lee, Ji-Won; Park, Ki-Ho; Chung, Jong-Hyuk; Kim, Su-Jung

    2011-11-01

    To evaluate and compare the effects of circumferential supracrestal fiberotomy in vivo (using diode, CO(2), and Er∶YAG lasers) on the morphology and chemical composition of the root surface. Forty healthy premolar teeth, intended for extraction for orthodontic reasons, were used in this study. Root surfaces were treated using different laser methods, as follows: (1) control; (2) Er∶YAG laser (2.94 µm, 100 mJ, 10 Hz); (3) diode laser (808 nm, 1.2 W, continuous wave); and (4) CO(2) laser (10.6 µm, 3 W, continuous wave). Subsequently, the teeth were removed and subjected to scanning electron microscopic (SEM) examination and energy dispersive x-ray (EDX) spectrometric analysis. SEM analysis indicated that no thermal changes, including melting or carbonization, were observed following the lasing procedures. EDX analysis showed that the laser procedures resulted in similar mineral contents (weight % of calcium and phosphate) as compared to those in the control group. Based on these findings, we concluded that laser-aided procedures, when used at appropriate laser settings, preserve the original morphology and chemical composition of cementum.

  10. Surface Characterization of Carbon Fiber Polymer Composites and Aluminum Alloys After Laser Interference Structuring

    NASA Astrophysics Data System (ADS)

    Sabau, Adrian S.; Greer, Clayton M.; Chen, Jian; Warren, Charles D.; Daniel, Claus

    2016-07-01

    The increasing use of carbon fiber-reinforced polymer matrix composites (CFPC) and aluminum alloys as lightweight materials in the automotive and aerospace industries demands enhanced surface preparation and control of surface morphology prior to joining. In this study, surfaces of both composite and aluminum were prepared for joining using an Nd:YAG laser in a two-beam interference setup, enabling the (1) structuring of the AL 5182 surface, (2) removal of the resin layer on top of carbon fibers, and (3) structuring of the carbon fibers. CFPC specimens of T700S carbon fiber, Prepreg—T83 epoxy, 5 ply thick, 0°/90° plaques were used. The effects of laser fluence, scanning speed, and number of shots-per-spot were investigated on the removal rate of the resin without an excessive damage of the fibers. Optical micrographs, 3D imaging, and scanning electron microscope imaging were used to study the effect of the laser processing on the surface morphology. It was found that an effective resin ablation and a low density of broken fibers for CFPC specimens was attained using laser fluences of 1-2 J/cm2 and number of 2-4 pulses per spot. A relatively large area of periodic line structures due to energy interference were formed on the aluminum surface at laser fluences of 12 J/cm2 and number of 4-6 pulses per spot.

  11. Prognostic significance of S-phase fractions in peritumoral invading zone analyzed by laser scanning cytometry in patients with high-grade glioma: A preliminary study.

    PubMed

    Nakajima, Syoichi; Morii, Ken; Takahashi, Hitoshi; Fujii, Yukihiko; Yamanaka, Ryuya

    2016-03-01

    The predominant characteristic of malignant glioma is the presence of invading tumor cells in the peritumoral zone. Distinguishing between tumor cells and normal cells in a peritumoral lesion is challenging. Therefore, the aim of the present study was to investigate the cell-cycle phase measurements of fixed paraffin-embedded specimens from the peritumoral invading zone of high-grade gliomas using laser scanning cytometry. A total of 12 high-grade gliomas (2 anaplastic astrocytomas and 10 glioblastomas) were studied. The tumor core and peritumoral invading zone of each tumor specimen were investigated. Tissue sections (50 µm) from the paraffin blocks were deparaffinized, rehydrated and enzymatically disintegrated, and the cells in suspension were stained with propidium iodide and placed on microscope slides. A slight trend for an increased S-phase fraction in the peritumoral invading zone compared with the tumor core was observed (P=0.24). Additionally, there was a trend for a decrease in the overall survival time of patients with increasing peritumoral invading zone S-phase fraction (P=0.12). These data suggest that laser scanning cytometry is a powerful and clinically relevant tool for the objective analysis of the cell cycle in malignant gliomas.

  12. Combination of intracellular staining of retrogradely labeled neurons and anterograde fluorescent tracing: use of the confocal laser scanning microscope.

    PubMed

    Shi, C; Cassell, M D

    1993-04-01

    This report describes a combined retrograde tracing, intracellular injection and anterograde fluorescence labeling method using the application of confocal laser scanning microscopy. By simultaneously viewing the morphology of identified projection neurons and the distribution of anterogradely labeled fibers and terminals, this approach allows accurate characterization of the anatomical relationships between these two elements. To demonstrate this approach, the retrograde tracer Fast Blue was injected into the bed nucleus of stria terminalis (BNST) and the anterograde tracer tetramethylrhodamine-conjugated dextran was injected into the insular cortex in adult rats. After one week survival time, the brains were fixed and sectioned on a vibratome. Individual BNST projecting neurons identified in the amygdaloid complex on 120 microns thick sections were intracellularly injected with Lucifer Yellow under visual control and analyzed with confocal laser scanning microscopy. The results demonstrate that images from very thin optical sections can clearly show potential synaptic contacts between anterograde labeling and intracellularly labeled projecting neurons. Stacked images from optical sections show, in very great detail, the morphology of projection neurons in three-dimensions. Compared to other methodological combinations, the present method provides a more simple and efficient means to trace three successive components of a putative neuron chain.

  13. Surface plasmon resonance and nonlinear optical behavior of pulsed laser-deposited semitransparent nanostructured copper thin films

    NASA Astrophysics Data System (ADS)

    Kesarwani, Rahul; Khare, Alika

    2018-06-01

    In this paper, surface plasmon resonance (SPR) and nonlinear optical properties of semitransparent nanostructured copper thin films fabricated on the glass substrate at 400 °C by pulsed laser deposition technique are reported. The thickness, linear absorption coefficient and linear refractive index of the films were measured by spectroscopic ellipsometer. The average particle size as measured via atomic force microscope was in the range of 12.84-26.02 nm for the deposition time ranging from 5 to 10 min, respectively. X-ray diffraction spectra revealed the formation of Cu (111) and Cu (200) planes. All these thin films exhibited broad SPR peak. The third-order optical nonlinearity of all the samples was investigated via modified z-scan technique using cw laser at a wavelength of 632.8 nm. The open aperture z-scan spectra of Cu thin film deposited for 5 min duration exhibited reverse saturation absorption whereas all the other samples displayed saturation absorption behavior. The nonlinear refractive index coefficient of these films showed a positive sign having the magnitude of the order of 10- 4 cm/W. The real and imaginary parts of susceptibilities were also calculated from the z-scan data and found to be of the order of 10- 6 esu.

  14. Skin welding using pulsed laser radiation and a dye

    NASA Astrophysics Data System (ADS)

    Fried, Nathaniel M.; Walsh, Joseph T., Jr.

    1998-07-01

    Previous skin welding studies have used continuous wave (CW) delivery of radiation. However, heat diffusion during irradiation prevents strong welds from being achieved without creating large zones of thermal damage to surrounding tissue. This damage may prevent normal wound healing. Strong welds and minimal thermal damage can be achieved by introducing a dye and delivering the radiation in a pulsed mode. Two-cm-long, full-thickness incisions were made in guinea pig skin. India ink was used as an absorber, and egg white albumin was used as an adhesive. A 5-mm-diameter spot of CW, 1.06-micrometer Nd:YAG laser radiation was scanned over the weld site, producing 100 millisecond pulses. The cooling time between scans and number of scans was varied. Thermal damage zones were measured using a transmission polarizing microscope to identify birefringence changes in tissue. Tensile strengths were measured using a tensiometer. For pulsed welding and long cooling times, weld strengths of 2.4 kg/cm2 were measured, and thermal damage to the epidermis was limited to approximately 500 micrometers. With CW welding, comparable weld strengths resulted in approximately 2700 micrometer of thermal damage. CW laser radiation weld strengths were only 0.6 kg/cm2 when thermal damage in the epidermis was limited to approximately 500 micrometers.

  15. Rotary-scanning optical resolution photoacoustic microscopy

    NASA Astrophysics Data System (ADS)

    Qi, Weizhi; Xi, Lei

    2016-10-01

    Optical resolution photoacoustic microscopy (ORPAM) is currently one of the fastest evolving photoacoustic imaging modalities. It has a comparable spatial resolution to pure optical microscopic techniques such as epifluorescence microscopy, confocal microscopy, and two-photon microscopy, but also owns a deeper penetration depth. In this paper, we report a rotary-scanning (RS)-ORPAM that utilizes a galvanometer scanner integrated with objective to achieve rotary laser scanning. A 15 MHz cylindrically focused ultrasonic transducer is mounted onto a motorized rotation stage to follow optical scanning traces synchronously. To minimize the loss of signal to noise ratio, the acoustic focus is precisely adjusted to reach confocal with optical focus. Black tapes and carbon fibers are firstly imaged to evaluate the performance of the system, and then in vivo imaging of vasculature networks inside the ears and brains of mice is demonstrated using this system.

  16. Laser Microdissection and Atmospheric Pressure Chemical Ionization Mass Spectrometry Coupled for Multimodal Imaging

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

    Lorenz, Matthias; Ovchinnikova, Olga S; Kertesz, Vilmos

    2013-01-01

    This paper describes the coupling of ambient laser ablation surface sampling, accomplished using a laser capture microdissection system, with atmospheric pressure chemical ionization mass spectrometry for high spatial resolution multimodal imaging. A commercial laser capture microdissection system was placed in close proximity to a modified ion source of a mass spectrometer designed to allow for sampling of laser ablated material via a transfer tube directly into the ionization region. Rhodamine 6G dye of red sharpie ink in a laser etched pattern as well as cholesterol and phosphatidylcholine in a cerebellum mouse brain thin tissue section were identified and imaged frommore » full scan mass spectra. A minimal spot diameter of 8 m was achieved using the 10X microscope cutting objective with a lateral oversampling pixel resolution of about 3.7 m. Distinguishing between features approximately 13 m apart in a cerebellum mouse brain thin tissue section was demonstrated in a multimodal fashion including co-registered optical and mass spectral chemical images.« less

  17. Femtosecond laser induced fixation of calcium alkali phosphate ceramics on titanium alloy bone implant material.

    PubMed

    Symietz, Christian; Lehmann, Erhard; Gildenhaar, Renate; Krüger, Jörg; Berger, Georg

    2010-08-01

    Femtosecond lasers provide a novel method of attaching bioceramic material to a titanium alloy, thereby improving the quality of bone implants. The ultrashort 30 fs laser pulses (790 nm wavelength) penetrate a thin dip-coated layer of fine ceramic powder, while simultaneously melting a surface layer of the underlying metal. The specific adjustment of the laser parameters (pulse energy and number of pulses per spot) avoids unnecessary melting of the bioactive calcium phosphate, and permits a defined thin surface melting of the metal, which in turn is not heated throughout, and therefore maintains its mechanical stability. It is essential to choose laser energy densities that correspond to the interval between the ablation fluences of both materials involved: about 0.1-0.4 Jcm(-2). In this work, we present the first results of this unusual technique, including laser ablation studies, scanning electron microscopy and optical microscope images, combined with EDX data. Copyright 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  18. Laser Surface Alloying of Copper, Manganese, and Magnesium with Pure Aluminum Substrate

    NASA Astrophysics Data System (ADS)

    Jiru, Woldetinsay G.; Sankar, M. Ravi; Dixit, Uday S.

    2016-03-01

    Laser surface alloying is one of the recent technologies used in the manufacturing sector for improving the surface properties of the metals. Light weight materials like aluminum alloys, titanium alloys, and magnesium alloys are used in the locomotive, aerospace, and structural applications. In the present work, an experimental study was conducted to improve the surface hardness of commercially pure aluminum plate. CO2 laser is used to melt pre-placed powders of pure copper, manganese, and magnesium. Microstructure of alloyed surface was analyzed using optical microscope. The best surface alloying was obtained at the optimum values of laser parameters, viz., laser power, scan speed, and laser beam diameter. In the alloyed region, microhardness increased from 30 HV0.5 to 430 HV0.5, while it was 60 HV0.5 in the heat-affected region. Tensile tests revealed some reduction in the strength and total elongation due to alloying. On the other hand, corrosion resistance improved.

  19. Material removal effect of microchannel processing by femtosecond laser

    NASA Astrophysics Data System (ADS)

    Zhang, Pan; Chen, Lei; Chen, Jianxiong; Tu, Yiliu

    2017-11-01

    Material processing using ultra-short-pulse laser is widely used in the field of micromachining, especially for the precision processing of hard and brittle materials. This paper reports a theoretical and experimental study of the ablation characteristics of a silicon wafer under micromachining using a femtosecond laser. The ablation morphology of the silicon wafer surface is surveyed by a detection test with an optical microscope. First, according to the relationship between the diameter of the ablation holes and the incident laser power, the ablation threshold of the silicon wafer is found to be 0.227 J/cm2. Second, the influence of various laser parameters on the size of the ablation microstructure is studied and the ablation morphology is analyzed. Furthermore, a mathematical model is proposed that can calculate the ablation depth per time for a given laser fluence and scanning velocity. Finally, a microchannel milling test is carried out on the micromachining center. The effectiveness and accuracy of the proposed models are verified by comparing the estimated depth to the actual measured results.

  20. Effect of Er,Cr:YSGG laser irradiation on eruption of rat mandibular incisor after disturbance of the enamel organ in the pulp.

    PubMed

    Masuda, Yoshiko Murakami; Hossain, Mozammal; Wang, Xiaogu; Matsuoka, Emi; Okano, Tomohiro; Matsumoto, Koukichi

    2006-09-01

    To investigate the efficacy of Er,Cr:YSGG (erbium,chromium:yttrium scandium gallium garnet) laser irradiation in root canal preparation and to evaluate its effect on eruption of rat incisors after disturbance of the enamel organ in the pulp, 20 canals of lower left incisor teeth were prepared by K-files followed by Er,Cr:YSGG laser irradiation, and 20 canals of right incisors were subjected to K-files only (control). At 1 week after irradiation, both sides of incisors erupted at the same level from the gingival margin. Histological findings showed that laser irradiation produced a slightly larger damage in the pulp than that of control. Scanning electron microscope observation revealed that laser-treated surface revealed a rough, irregular, and very clean surface; there was almost no evidence of debris or smear layer, and dentinal tubules were opened. Adequate power of Er,Cr:YSGG laser irradiation is effective in root canal preparation without disturbance of the eruption.

  1. Laser Cladding of Composite Bioceramic Coatings on Titanium Alloy

    NASA Astrophysics Data System (ADS)

    Xu, Xiang; Han, Jiege; Wang, Chunming; Huang, Anguo

    2016-02-01

    In this study, silicon nitride (Si3N4) and calcium phosphate tribasic (TCP) composite bioceramic coatings were fabricated on a Ti6Al4V (TC4) alloy using Nd:YAG pulsed laser, CO2 CW laser, and Semiconductor CW laser. The surface morphology, cross-sectional microstructure, mechanical properties, and biological behavior were carefully investigated. These investigations were conducted employing scanning electron microscope, energy-dispersive x-ray spectroscopy, and other methodologies. The results showed that both Si3N4 and Si3N4/TCP composite coatings were able to form a compact bonding interface between the coating and the substrate by using appropriate laser parameters. The coating layers were dense, demonstrating a good surface appearance. The bioceramic coatings produced by laser cladding have good mechanical properties. Compared with that of the bulk material, microhardness of composite ceramic coatings on the surface significantly increased. In addition, good biological activity could be obtained by adding TCP into the composite coating.

  2. Enhanced weak-signal sensitivity in two-photon microscopy by adaptive illumination.

    PubMed

    Chu, Kengyeh K; Lim, Daryl; Mertz, Jerome

    2007-10-01

    We describe a technique to enhance both the weak-signal relative sensitivity and the dynamic range of a laser scanning optical microscope. The technique is based on maintaining a fixed detection power by fast feedback control of the illumination power, thereby transferring high measurement resolution to weak signals while virtually eliminating the possibility of image saturation. We analyze and demonstrate the benefits of adaptive illumination in two-photon fluorescence microscopy.

  3. Nanospectrofluorometry inside single living cell by scanning near-field optical microscopy

    NASA Astrophysics Data System (ADS)

    Lei, F. H.; Shang, G. Y.; Troyon, M.; Spajer, M.; Morjani, H.; Angiboust, J. F.; Manfait, M.

    2001-10-01

    Near-field fluorescence spectra with subdiffraction limit spatial resolution have been taken in the proximity of mitochondrial membrane inside breast adenocarcinoma cells (MCF7) treated with the fluorescent dye (JC-1) by using a scanning near-field optical microscope coupled with a confocal laser microspectrofluorometer. The probe-sample distance control is based on a piezoelectric bimorph shear force sensor having a static spring constant k=5 μN/nm and a quality factor Q=40 in a physiological medium of viscosity η=1.0 cp. The sensitivity of the force sensor has been tested by imaging a MCF7 cell surface.

  4. TEM and STEM Studies on the Cross-sectional Morphologies of Dual-/Tri-layer Broadband SiO2 Antireflective Films

    NASA Astrophysics Data System (ADS)

    Wang, Shuangyue; Yan, Hongwei; Li, Dengji; Qiao, Liang; Han, Shaobo; Yuan, Xiaodong; Liu, Wei; Xiang, Xia; Zu, Xiaotao

    2018-02-01

    Dual-layer and tri-layer broadband antireflective (AR) films with excellent transmittance were successfully fabricated using base-/acid-catalyzed mixed sols and propylene oxide (PO) modified silica sols. The sols and films were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), transmission electron microscope (TEM), and scanning transmission electron microscope (STEM). FTIR and TEM results suggest that the PO molecules were covalently bonded to the silica particles and the bridge structure existing in PO modified silica sol is responsible for the low density of the top layer. The density ratio between different layers was measured by cross-sectional STEM, and the results are 1.69:1 and 2.1:1.7:1 from bottom-layer to top-layer for dual-layer and tri-layer films, respectively. The dual-layer film demonstrates good stability with 99.8% at the central wavelength of 351 nm and nearly 99.5% at the central wavelength of 1053 nm in laser system, and for the tri-layer AR film, the maximum transmittance reached nearly 100% at both the central wavelengths of 527 and 1053 nm.

  5. Thermally-induced voltage alteration for integrated circuit analysis

    DOEpatents

    Cole, Jr., Edward I.

    2000-01-01

    A thermally-induced voltage alteration (TIVA) apparatus and method are disclosed for analyzing an integrated circuit (IC) either from a device side of the IC or through the IC substrate to locate any open-circuit or short-circuit defects therein. The TIVA apparatus uses constant-current biasing of the IC while scanning a focused laser beam over electrical conductors (i.e. a patterned metallization) in the IC to produce localized heating of the conductors. This localized heating produces a thermoelectric potential due to the Seebeck effect in any conductors with open-circuit defects and a resistance change in any conductors with short-circuit defects, both of which alter the power demand by the IC and thereby change the voltage of a source or power supply providing the constant-current biasing. By measuring the change in the supply voltage and the position of the focused and scanned laser beam over time, any open-circuit or short-circuit defects in the IC can be located and imaged. The TIVA apparatus can be formed in part from a scanning optical microscope, and has applications for qualification testing or failure analysis of ICs.

  6. Investigation of Carbon Fiber Reinforced Plastics Machining Using 355 nm Picosecond Pulsed Laser

    NASA Astrophysics Data System (ADS)

    Hu, Jun; Zhu, Dezhi

    2018-06-01

    Carbon fiber reinforced plastics (CFRP) has been widely used in the aircraft industry and automobile industry owing to its superior properties. In this paper, a Nd:YVO4 picosecond pulsed system emitting at 355 nm has been used for CFRP machining experiments to determine optimum milling conditions. Milling parameters including laser power, milling speed and hatch distance were optimized by using box-behnken design of response surface methodology (RSM). Material removal rate was influenced by laser beam overlap ratio which affects mechanical denudation. The results in heat affected zones (HAZ) and milling quality were discussed through the machined surface observed with scanning electron microscope. A re-focusing technique based on the experiment with different focal planes was proposed and milling mechanism was also analyzed in details.

  7. Surface characterization of stainless HP-40 steel using laser induced μ-breakdown spectroscopy (μ -LIBS)

    NASA Astrophysics Data System (ADS)

    Pinto, M.; Calderón, X.; Mejía Ospino, E.; Cabanzo, R.; Poveda, Juan C.

    2016-02-01

    In the present study, optical microscopy in stereoscopic mode coupled to laser- induced p-breakdown spectroscopy (μ-LIBS) was applied for analysing HP-40 steel samples. microLIBS (μ-LIBS) is a new growing area that employs low energy laser pulses for the generation of plasma emission, which allow the realization of localized microanalysis [1]. This new LIBS instrument was used for the surface characterization of the steel samples in the spectral range from 356 to 401nm. Elements such as Cr, Ni, Fe, Nb, Pb, Mo, C, Mn and Si in the steel samples were investigated. The results allowed the construction of elemental distribution profiles of the samples. Complementary the HP-40 steel samples were superficially characterized by Scanning Electron Microscope (SEM).

  8. Curved laser microjet in near field.

    PubMed

    Kotlyar, Victor V; Stafeev, Sergey S; Kovalev, Alexey A

    2013-06-20

    With the use of the finite-difference time-domain-based simulation and a scanning near-field optical microscope that has a metal cantilever tip, the diffraction of a linearly polarized plane wave of wavelength λ by a glass corner step of height 2λ is shown to generate a low divergence laser jet of a root-parabolic form: over a distance of 4.7λ on the optical axis, the beam path is shifted by 2.1λ. The curved laser jet of the FWHM length depth of focus=9.5λ has the diameter FWHM=1.94λ over the distance 5.5λ, and the intensity maximum is 5 times higher than the incident wave intensity. The discrepancy between the analytical and the experimental results amounts to 11%.

  9. Silver/oxygen depth profile in coins by using laser ablation, mass quadrupole spectrometer and X-rays fluorescence

    NASA Astrophysics Data System (ADS)

    Cutroneo, M.; Torrisi, L.; Caridi, F.; Sayed, R.; Gentile, C.; Mondio, G.; Serafino, T.; Castrizio, E. D.

    2013-05-01

    Silver coins belonging to different historical periods were investigated to determine the Ag/O atomic ratio depth profiles. Laser ablation has been employed to remove, in high vacuum, the first superficial layers of the coins. Mass quadrupole spectrometry has been used to detect the Ag and the O atomic elements vaporized from the coin surface. The depth profile allowed to determine the thickness of the oxidation layer indicating that, in general, it is high in old coins. A complementary technique, using scanning electron microscope and the associated XRF microprobe, have been devoted to confirm the measurements of Ag/O atomic ratio measured with the laser-coupled mass spectrometry. The oxidation layer thicknesses range between about 25 and 250 microns.

  10. Ultrashort pulsed laser (USPL) application in dentistry: basic investigations of ablation rates and thresholds on oral hard tissue and restorative materials.

    PubMed

    Schelle, Florian; Polz, Sebastian; Haloui, Hatim; Braun, Andreas; Dehn, Claudia; Frentzen, Matthias; Meister, Jörg

    2014-11-01

    Modern ultrashort pulse lasers with scanning systems provide a huge set of parameters affecting the suitability for dental applications. The present study investigates thresholds and ablation rates of oral hard tissues and restorative materials with a view towards a clinical application system. The functional system consists of a 10 W Nd:YVO4 laser emitting pulses with a duration of 8 ps at 1,064 nm. Measurements were performed on dentin, enamel, ceramic, composite, and mammoth ivory at a repetition rate of 500 kHz. By employing a scanning system, square-shaped cavities with an edge length of 1 mm were created. Ablation threshold and rate measurements were assessed by variation of the applied fluence. Examinations were carried out employing a scanning electron microscope and optical profilometer. Irradiation time was recorded by the scanner software in order to calculate the overall ablated volume per time. First high power ablation rate measurements were performed employing a laser source with up to 50 W. Threshold values in the range of 0.45 J/cm(2) (composite) to 1.54 J/cm(2) (enamel) were observed. Differences between any two materials are statistically significant (p < 0.05). Preparation speeds up to 37.53 mm(3)/min (composite) were achieved with the 10 W laser source and differed statistically significant for any two materials (p < 0.05) with the exception of dentin and mammoth ivory (p > 0.05). By employing the 50 W laser source, increased rates up to ∼50 mm(3)/min for dentin were obtained. The results indicate that modern USPL systems provide sufficient ablation rates to be seen as a promising technology for dental applications.

  11. Image correlation microscopy for uniform illumination.

    PubMed

    Gaborski, T R; Sealander, M N; Ehrenberg, M; Waugh, R E; McGrath, J L

    2010-01-01

    Image cross-correlation microscopy is a technique that quantifies the motion of fluorescent features in an image by measuring the temporal autocorrelation function decay in a time-lapse image sequence. Image cross-correlation microscopy has traditionally employed laser-scanning microscopes because the technique emerged as an extension of laser-based fluorescence correlation spectroscopy. In this work, we show that image correlation can also be used to measure fluorescence dynamics in uniform illumination or wide-field imaging systems and we call our new approach uniform illumination image correlation microscopy. Wide-field microscopy is not only a simpler, less expensive imaging modality, but it offers the capability of greater temporal resolution over laser-scanning systems. In traditional laser-scanning image cross-correlation microscopy, lateral mobility is calculated from the temporal de-correlation of an image, where the characteristic length is the illuminating laser beam width. In wide-field microscopy, the diffusion length is defined by the feature size using the spatial autocorrelation function. Correlation function decay in time occurs as an object diffuses from its original position. We show that theoretical and simulated comparisons between Gaussian and uniform features indicate the temporal autocorrelation function depends strongly on particle size and not particle shape. In this report, we establish the relationships between the spatial autocorrelation function feature size, temporal autocorrelation function characteristic time and the diffusion coefficient for uniform illumination image correlation microscopy using analytical, Monte Carlo and experimental validation with particle tracking algorithms. Additionally, we demonstrate uniform illumination image correlation microscopy analysis of adhesion molecule domain aggregation and diffusion on the surface of human neutrophils.

  12. A study of the mechanism of laser welding defects in low thermal expansion superalloy GH909

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

    Yan, Fei; Wang, Chunming, E-mail: yanxiangfei225@163.com; Wang, Yajun

    2013-04-15

    In this paper, we describe experimental laser welding of low-thermal-expansion superalloy GH909. The main welding defects of GH909 by laser in the weld are liquation cracks and porosities, including hydrogen and carbon monoxide porosity. The forming mechanism of laser welding defects was investigated. This investigation was conducted using an optical microscope, scanning electron microscope, energy diffraction spectrum, X-ray diffractometer and other methodologies. The results demonstrated that porosities appearing in the central weld were related to incomplete removal of oxide film on the surface of the welding samples. The porosities produced by these bubbles were formed as a result of residualmore » hydrogen or oxygenium in the weld. These elements failed to escape from the weld since laser welding has both a rapid welding speed and cooling rate. The emerging crack in the heat affected zone is a liquation crack and extends along the grain boundary as a result of composition segregation. Laves–Ni{sub 2}Ti phase with low melting point is a harmful phase, and the stress causes grain boundaries to liquefy, migrate and even crack. Removing the oxides on the surface of the samples before welding and carefully controlling technological parameters can reduce welding defects and improve formation of the GH909 alloy weld. - Highlights: ► It is a new process for the forming of GH909 alloy via laser welding. ► The forming mechanism of laser welding defects in GH909 has been studied. ► It may be a means to improve the efficiency of aircraft engine production.« less

  13. Use of the erbium, chromium:yttrium-scandium-gallium-garnet laser on human enamel tissues. Influence of the air-water spray on the laser-tissue interaction: scanning electron microscope evaluations.

    PubMed

    Olivi, Giovanni; Angiero, Francesca; Benedicenti, Stefano; Iaria, Giuseppe; Signore, Antonio; Kaitsas, Vassilios

    2010-11-01

    The study investigated the influence of varying amounts of air/water spray and the energy used by an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) 2,780 nm laser when treating dental tissues. The morphological effects produced by the laser interaction on healthy human enamel were evaluated by scanning electron microscopy (SEM). The vestibular and lingual surfaces of ten molars were treated with laser at different power settings; each surface was subdivided into cervical, median, and occlusal parts and treated with different proportions of water spray; the series contained 60 tooth portions. Treatment differed in terms of power setting and air/water percentage. All specimens were then subjected to dehydration and metallisation. At SEM evaluation, the classic aspect of laser-treated enamel was visible: grooves, flakes, shelves and sharp edges, indicative of micro-explosion rather than melting. Vaporisation of the tissue created a clear delimitation from surrounding healthy tissue, with partial respect to the prismatic structure of the treated enamel. The aspect of the enamel was rarely type 1 Silverstone but more frequently type 2 or 3, with prismatic structure not respected and/or completely disordered. These morphological differences appeared to be correlated with the inclination of the laser beam aimed at the enamel prisms and with the percentage of air/water used. The laser system analysed showed itself to be effective at removing human dental enamel. The results appeared to be closely correlated with the variation of the percentage of the laser's water-air spray.

  14. Fabrication of hydrophobic structures on coronary stent surface based on direct three-beam laser interference lithography

    NASA Astrophysics Data System (ADS)

    Gao, Long-yue; Zhou, Wei-qi; Wang, Yuan-bo; Wang, Si-qi; Bai, Chong; Li, Shi-ming; Liu, Bin; Wang, Jun-nan; Cui, Cheng-kun; Li, Yong-liang

    2016-05-01

    To solve the problems with coronary stent implantation, coronary artery stent surface was directly modified by three-beam laser interference lithography through imitating the water-repellent surface of lotus leaf, and uniform micro-nano structures with the controllable period were fabricated. The morphological properties and contact angle (CA) of the microstructure were measured by scanning electron microscope (SEM) and CA system. The water repellency of stent was also evaluated by the contact and then separation between the water drop and the stent. The results show that the close-packed concave structure with the period of about 12 μm can be fabricated on the stent surface with special parameters (incident angle of 3°, laser energy density of 2.2 J·cm-2 and exposure time of 80 s) by using the three-beam laser at 1 064 nm, and the structure has good water repellency with CA of 120°.

  15. Pulsed Nd-YAG laser in endodontics

    NASA Astrophysics Data System (ADS)

    Ragot-Roy, Brigitte; Severin, Claude; Maquin, Michel

    1994-12-01

    The purpose of this study was to establish an operative method in endodontics. The effect of a pulsed Nd:YAG laser on root canal dentin has been examined with a scanning electron microscope. Our first experimentation was to observe the impacts carried out perpendicularly to root canal surface with a 200 micrometers fiber optic in the presence of dye. Secondarily, the optical fiber was used as an endodontic instrument with black dye. The irradiation was performed after root canal preparation (15/100 file or 40/100 file) or directly into the canal. Adverse effects are observed. The results show that laser irradiation on root canal dentin surfaces induces a nonhomogeneous modified dentin layer, melted and resolidified dentin closed partially dentinal tubules. The removal of debris is not efficient enough. The laser treatment seems to be indicated only for endodontic and periapical spaces sterilization after conventional root canal preparation.

  16. Study on ceramic coating on the enamel surface using a carbon dioxide laser.

    PubMed

    Nihei, Tomotaro; Kurata, Shigeaki; Ohashi, Katsura; Umemoto, Kozo; Teranaka, Toshio

    2011-01-01

    The aims of this study were to evaluate a new restorative method using a carbon dioxide laser (CO(2)-laser) and to evaluate the acid resistance of teeth. Experimental calcium phosphate glass (CPG) powder and two low melting point ceramics (Finesse and zirconium silicate) were fused to enamel surfaces using a CO(2)-laser at an irradiation intensity of 1.0 watt for 30 seconds with a beam size of 0.49 mm at the focal point. The treated teeth were observed with a scanning electron microscope, and the acid resistance of the treated enamel surfaces was evaluated. The CPG fused successfully to the enamel surface, and the treated enamel surface showed high acid resistance compared with the low melting point ceramics and the non-irradiated surfaces. This system may lead to the development of new restorative methods that do not require the use of bonding agents.

  17. Experimental investigation on the electrical contact behavior of rolling contact connector.

    PubMed

    Chen, Junxing; Yang, Fei; Luo, Kaiyu; Zhu, Mingliang; Wu, Yi; Rong, Mingzhe

    2015-12-01

    Rolling contact connector (RCC) is a new technology utilized in high performance electric power transfer systems with one or more rotating interfaces, such as radars, satellites, wind generators, and medical computed tomography machines. Rolling contact components are used in the RCC instead of traditional sliding contacts to transfer electrical power and/or signal. Since the requirement of the power transmission is increasing in these years, the rolling electrical contact characteristics become more and more important for the long-life design of RCC. In this paper, a typical form of RCC is presented. A series of experimental work are carried out to investigate the rolling electrical contact characteristics during its lifetime. The influence of a variety of factors on the electrical contact degradation behavior of RCC is analyzed under both vacuum and air environment. Based on the surface morphology and elemental composition changes in the contact zone, which are assessed by field emission scanning electron microscope and confocal laser scanning microscope, the mechanism of rolling electrical contact degradation is discussed.

  18. Parallel detection experiment of fluorescence confocal microscopy using DMD.

    PubMed

    Wang, Qingqing; Zheng, Jihong; Wang, Kangni; Gui, Kun; Guo, Hanming; Zhuang, Songlin

    2016-05-01

    Parallel detection of fluorescence confocal microscopy (PDFCM) system based on Digital Micromirror Device (DMD) is reported in this paper in order to realize simultaneous multi-channel imaging and improve detection speed. DMD is added into PDFCM system, working to take replace of the single traditional pinhole in the confocal system, which divides the laser source into multiple excitation beams. The PDFCM imaging system based on DMD is experimentally set up. The multi-channel image of fluorescence signal of potato cells sample is detected by parallel lateral scanning in order to verify the feasibility of introducing the DMD into fluorescence confocal microscope. In addition, for the purpose of characterizing the microscope, the depth response curve is also acquired. The experimental result shows that in contrast to conventional microscopy, the DMD-based PDFCM system has higher axial resolution and faster detection speed, which may bring some potential benefits in the biology and medicine analysis. SCANNING 38:234-239, 2016. © 2015 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

  19. Anisotropic excitation of surface plasmon polaritons on a metal film by a scattering-type scanning near-field microscope with a non-rotationally-symmetric probe tip

    NASA Astrophysics Data System (ADS)

    Walla, Frederik; Wiecha, Matthias M.; Mecklenbeck, Nicolas; Beldi, Sabri; Keilmann, Fritz; Thomson, Mark D.; Roskos, Hartmut G.

    2018-01-01

    We investigated the excitation of surface plasmon polaritons on gold films with the metallized probe tip of a scattering-type scanning near-field optical microscope (s-SNOM). The emission of the polaritons from the tip, illuminated by near-infrared laser radiation, was found to be anisotropic and not circularly symmetric as expected on the basis of literature data. We furthermore identified an additional excitation channel via light that was reflected off the tip and excited the plasmon polaritons at the edge of the metal film. Our results, while obtained for a non-rotationally-symmetric type of probe tip and thus specific for this situation, indicate that when an s-SNOM is employed for the investigation of plasmonic structures, the unintentional excitation of surface waves and anisotropic surface wave propagation must be considered in order to correctly interpret the signatures of plasmon polariton generation and propagation.

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

    Chen, Junxing; Yang, Fei, E-mail: yfei2007@mail.xjtu.edu.cn; Luo, Kaiyu

    Rolling contact connector (RCC) is a new technology utilized in high performance electric power transfer systems with one or more rotating interfaces, such as radars, satellites, wind generators, and medical computed tomography machines. Rolling contact components are used in the RCC instead of traditional sliding contacts to transfer electrical power and/or signal. Since the requirement of the power transmission is increasing in these years, the rolling electrical contact characteristics become more and more important for the long-life design of RCC. In this paper, a typical form of RCC is presented. A series of experimental work are carried out to investigatemore » the rolling electrical contact characteristics during its lifetime. The influence of a variety of factors on the electrical contact degradation behavior of RCC is analyzed under both vacuum and air environment. Based on the surface morphology and elemental composition changes in the contact zone, which are assessed by field emission scanning electron microscope and confocal laser scanning microscope, the mechanism of rolling electrical contact degradation is discussed.« less

  1. Formation of nanograting in fused silica by temporally delayed femtosecond double-pulse irradiation

    NASA Astrophysics Data System (ADS)

    Wang, Haodong; Song, Juan; Li, Qin; Zeng, Xianglong; Dai, Ye

    2018-04-01

    A 1 kHz femtosecond double-pulse sequence irradiation is used to study the temporal evolution of nanograting in fused silica by controlling the delay times and polarization combinations of two independent beams from a Mach–Zehnder interferometer. A lateral laser-scan experiment with speed at 5 µm s‑1 and each pulse energy of 1 µJ is firstly performed with the delay time from sub-picosecond to 10 ps, and then the written nanostructures are systematically studied under a cross-polarized microscope because the intensity of birefringence signal nearly corresponds to optical retardance and development level of the induced nanograting. The trend shows that the induced nanogratings can continue developing with a decrease of delay time in the case of the linear polarization pulse arriving before. In another vertical laser-scan experiment at the same speed and pulse energy, the morphologies of nanogratings embedded in the lines are characterized by scanning electron microscope after mechanical polishing and chemical etching. The self-organized patterns have a commonly spatial period of 200–300 nm and the orientation is always perpendicular to the polarization of the first laser pulse, and the second pulse in each sequence seems to promote the as-formed nanograting developing further even if the polarized direction is different from the previous pulse. These new findings verify again that a localized memory effect can make positive feedback to reinforce the patterned nanostripes. In that process, the impact ionization from the seed electrons left by the first pulse excitation and the photoionization of self-trapped excitons with lower ionization threshold results in an increase of the re-excited carriers during the second pulse irradiation and the subsequent development of the as-formed nanograting. Our result provides further proofs for understanding the physical mechanism of nanograting strongly connection with the interplay on multiple ionization channels.

  2. Application of a flexible CO(2) laser fiber for neurosurgery: laser-tissue interactions.

    PubMed

    Ryan, Robert W; Wolf, Tamir; Spetzler, Robert F; Coons, Stephen W; Fink, Yoel; Preul, Mark C

    2010-02-01

    The CO(2) laser has an excellent profile for use in neurosurgery. Its high absorption in water results in low thermal spread, sparing adjacent tissue. Use of this laser has been limited to line-of-sight applications because no solid fiber optic cables could transmit its wavelength. Flexible photonic bandgap fiber technology enables delivery of CO(2) laser energy through a flexible fiber easily manipulated in a handheld device. The authors examined and compared the first use of this CO(2) laser fiber to conventional methods for incising neural tissue. Carbon dioxide laser energy was delivered in pulsed or continuous wave settings for different power settings, exposure times, and distances to cortical tissue of 6 anesthetized swine. Effects of CO(2) energy on the tissue were compared with bipolar cautery using a standard pial incision technique, and with scalpel incisions without cautery. Tissue was processed for histological analysis (using H & E, silver staining, and glial fibrillary acidic protein immunohistochemistry) and scanning electron microscopy, and lesion measurements were made. Light microscopy and scanning electron microscopy revealed laser incisions of consistent shape, with central craters surrounded by limited zones of desiccated and edematous tissue. Increased laser power resulted in deeper but not significantly wider incisions. Bipolar cautery lesions showed desiccated and edematous zones but did not incise the pia, and width increased more than depth with higher power. Incisions made without using cautery produced hemorrhage but minimal adjacent tissue damage. The photonic bandgap fiber CO(2) laser produced reliable cortical incisions, adjustable over a range of settings, with minimal adjacent thermal tissue damage. Ease of application under the microscope suggests this laser system has reached true practicality for neurosurgery.

  3. Clinical and laboratory applications of slide-based cytometry with the LSC, SFM, and the iCYTE imaging cytometer instruments

    NASA Astrophysics Data System (ADS)

    Bocsi, Jozsef; Luther, Ed; Mittag, Anja; Jensen, Ingo; Sack, Ulrich; Lenz, Dominik; Trezl, Lajos; Varga, Viktor S.; Molnar, Beea; Tarnok, Attila

    2004-06-01

    Background: Slide based cytometry (SBC) is a technology for the rapid stoichiometric analysis of cells fixed to surfaces. Its applications are highly versatile and ranges from the clinics to high throughput drug discovery. SBC is realized in different instruments such as the Laser Scanning Cytometer (LSC) and Scanning Fluorescent Microscope (SFM) and the novel inverted microscope based iCyte image cytometer (Compucyte Corp.). Methods: Fluorochrome labeled specimens were immobilized on microscopic slides. They were placed on a conventional fluorescence microscope and analyzed by photomultiplayers or digital camera. Data comparable to flow cytometry were generated. In addition, each individual event could be visualized. Applications: The major advantage of instruments is the combination of two features: a) the minimal sample volume needed, and b) the connection of fluorescence data and morphological information. Rare cells were detected, frequency of apoptosis by myricetin formaldehyde and H2O2 mixtures was determined;. Conclusion: LSC, SFM and the novel iCyte have a wide spectrum of applicability in SBC and can be introduced as a standard technology for multiple settings. In addition, the iCyte and SFM instrument is suited for high throughput screening by automation and may be in future adapted to telepathology due to their high quality images. (This study was supported by the IZKF-Leipzig, Germany and T 034245 OTKA, Hungary)

  4. Visualization of Neutrophil Extracellular Traps and Fibrin Meshwork in Human Fibrinopurulent Inflammatory Lesions: III. Correlative Light and Electron Microscopic Study

    PubMed Central

    Onouchi, Takanori; Shiogama, Kazuya; Mizutani, Yasuyoshi; Takaki, Takashi; Tsutsumi, Yutaka

    2016-01-01

    Neutrophil extracellular traps (NETs) released from dead neutrophils at the site of inflammation represent webs of neutrophilic DNA stretches dotted with granule-derived antimicrobial proteins, including lactoferrin, and play important roles in innate immunity against microbial infection. We have shown the coexistence of NETs and fibrin meshwork in varied fibrinopurulent inflammatory lesions at both light and electron microscopic levels. In the present study, correlative light and electron microscopy (CLEM) employing confocal laser scanning microscopy and scanning electron microscopy was performed to bridge light and electron microscopic images of NETs and fibrin fibrils in formalin-fixed, paraffin-embedded, autopsied lung sections of legionnaire’s pneumonia. Lactoferrin immunoreactivity and 4'-6-diamidino-2-phenylindole (DAPI) reactivity were used as markers of NETs, and fibrin was probed by fibrinogen gamma chain. Of note is that NETs light microscopically represented as lactoferrin and DAPI-colocalized dots, 2.5 μm in diameter. CLEM gave super-resolution images of NETs and fibrin fibrils: “Dotted” NETs were ultrastructurally composed of fine filaments and masses of 58 nm-sized globular materials. A fibrin fibril consisted of clusters of smooth-surfaced filaments. NETs filaments (26 nm in diameter) were significantly thinner than fibrin filaments (295 nm in diameter). Of note is that CLEM was applicable to formalin-fixed, paraffin-embedded sections of autopsy material. PMID:27917008

  5. Biological applications of an LCoS-based programmable array microscope (PAM)

    NASA Astrophysics Data System (ADS)

    Hagen, Guy M.; Caarls, Wouter; Thomas, Martin; Hill, Andrew; Lidke, Keith A.; Rieger, Bernd; Fritsch, Cornelia; van Geest, Bert; Jovin, Thomas M.; Arndt-Jovin, Donna J.

    2007-02-01

    We report on a new generation, commercial prototype of a programmable array optical sectioning fluorescence microscope (PAM) for rapid, light efficient 3D imaging of living specimens. The stand-alone module, including light source(s) and detector(s), features an innovative optical design and a ferroelectric liquid-crystal-on-silicon (LCoS) spatial light modulator (SLM) instead of the DMD used in the original PAM design. The LCoS PAM (developed in collaboration with Cairn Research, Ltd.) can be attached to a port of a(ny) unmodified fluorescence microscope. The prototype system currently operated at the Max Planck Institute incorporates a 6-position high-intensity LED illuminator, modulated laser and lamp light sources, and an Andor iXon emCCD camera. The module is mounted on an Olympus IX71 inverted microscope with 60-150X objectives with a Prior Scientific x,y, and z high resolution scanning stages. Further enhancements recently include: (i) point- and line-wise spectral resolution and (ii) lifetime imaging (FLIM) in the frequency domain. Multiphoton operation and other nonlinear techniques should be feasible. The capabilities of the PAM are illustrated by several examples demonstrating single molecule as well as lifetime imaging in live cells, and the unique capability to perform photoconversion with arbitrary patterns and high spatial resolution. Using quantum dot coupled ligands we show real-time binding and subsequent trafficking of individual ligand-growth factor receptor complexes on and in live cells with a temporal resolution and sensitivity exceeding those of conventional CLSM systems. The combined use of a blue laser and parallel LED or visible laser sources permits photoactivation and rapid kinetic analysis of cellular processes probed by photoswitchable visible fluorescent proteins such as DRONPA.

  6. Nanodissection of human chromosomes and ultraprecise eye surgery with nanojoule near-infrared femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Koenig, Karsten; Riemann, Iris; Krauss, Oliver; Fritzsche, Wolfgang

    2002-04-01

    Nanojoule and sub-nanojoule 80 MHz femtosecond laser pulses at 750-850 nm of a compact titanium:sapphire laser have been used for highly precise nanoprocessing of DNA as well as of intracellular and intratissue compartments. In particular, a mean power between 15 mW and 100 mW, 170 fs pulse width, submicron distance of illumination spots and microsecond beam dwell times on spots have been used for multiphoton- mediated nanoprocessing of human chromosomes, brain and ocular intrastromal tissue. By focusing the laser beam with high numerical aperture focusing optics of the laser scan system femt-O-cut and of modified multiphoton scanning microscopes to diffraction-limited spots and TW/cm2 light intensities, precise submicron holes and cuts have been processed by single spot exposure and line scans. A minimum FWHM cut size below 70 nm during the partial dissection of the human chromosome 3 was achieved. Complete chromosome dissection could be performed with FWHM cut sizes below 200 nm. Intracellular chromosome dissection was possible. Intratissue processing in depths of 50 - 100micrometers and deeper with a precision of about 1micrometers including cuts through a nuclei of a single intratissue cell without destructive photo-disruption effects to surrounding tissue layers have been demonstrated in brain and eye tissues. The femt-O-cut system includes a diagnostic system for optical tomography with submicron resolution based on multiphoton- excited autofluorescence imaging (MAI) and second harmonic generation. This system was used to localize the intracellular and intratissue targets and to control the effects of nanoprocessing. These studies show, that in contrast to conventional approaches of material processing with amplified femtosecond laser systems and (mu) J pulse energies, nanoprocessing of materials including biotissues can be performed with nJ and sub-nJ high repetition femtosecond laser pulses of turn-key compact lasers without collateral damage. Potential applications include highly precise cell and embryo surgery, gene diagnostics and gene therapy, intrastromal refractive surgery, cancer therapy and brain surgery.

  7. (LMRG): Microscope Resolution, Objective Quality, Spectral Accuracy and Spectral Un-mixing

    PubMed Central

    Bayles, Carol J.; Cole, Richard W.; Eason, Brady; Girard, Anne-Marie; Jinadasa, Tushare; Martin, Karen; McNamara, George; Opansky, Cynthia; Schulz, Katherine; Thibault, Marc; Brown, Claire M.

    2012-01-01

    The second study by the LMRG focuses on measuring confocal laser scanning microscope (CLSM) resolution, objective lens quality, spectral imaging accuracy and spectral un-mixing. Affordable test samples for each aspect of the study were designed, prepared and sent to 116 labs from 23 countries across the globe. Detailed protocols were designed for the three tests and customized for most of the major confocal instruments being used by the study participants. One protocol developed for measuring resolution and objective quality was recently published in Nature Protocols (Cole, R. W., T. Jinadasa, et al. (2011). Nature Protocols 6(12): 1929–1941). The first study involved 3D imaging of sub-resolution fluorescent microspheres to determine the microscope point spread function. Results of the resolution studies as well as point spread function quality (i.e. objective lens quality) from 140 different objective lenses will be presented. The second study of spectral accuracy looked at the reflection of the laser excitation lines into the spectral detection in order to determine the accuracy of these systems to report back the accurate laser emission wavelengths. Results will be presented from 42 different spectral confocal systems. Finally, samples with double orange beads (orange core and orange coating) were imaged spectrally and the imaging software was used to un-mix fluorescence signals from the two orange dyes. Results from 26 different confocal systems will be summarized. Time will be left to discuss possibilities for the next LMRG study.

  8. Chondrocytes provide a model for in-situ confocal microscopy and 3D reconstructions

    NASA Astrophysics Data System (ADS)

    Hirsch, Michelle S.; Svoboda, Kathy K. H.

    1994-04-01

    Hyaline cartilage is composed of chondrocytes that reside in lacunae surrounded by extracellular matrix molecules. Microscopic and histochemical features of cartilage have been studied with many techniques. Many of these techniques can be time consuming and may alter natural cartilage characteristics. In addition, the orientation and order of sectioned tissue must be maintained to create 3D reconstructions. We show that confocal laser scanning microscopy may replace traditional methods for studying cartilage.

  9. Visualizing tributyltin (TBT) in bacterial aggregates by specific rhodamine-based fluorescent probes.

    PubMed

    Jin, Xilang; Hao, Likai; She, Mengyao; Obst, Martin; Kappler, Andreas; Yin, Bing; Liu, Ping; Li, Jianli; Wang, Lanying; Shi, Zhen

    2015-01-01

    Here we present the first examples of fluorescent and colorimetric probes for microscopic TBT imaging. The fluorescent probes are highly selective and sensitive to TBT and have successfully been applied for imaging of TBT in bacterial Rhodobacter ferrooxidans sp. strain SW2 cell-EPS-mineral aggregates and in cell suspensions of the marine cyanobacterium Synechococcus PCC 7002 by using confocal laser scanning microscopy. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Adaptive thresholding image series from fluorescence confocal scanning laser microscope using orientation intensity profiles

    NASA Astrophysics Data System (ADS)

    Feng, Judy J.; Ip, Horace H.; Cheng, Shuk H.

    2004-05-01

    Many grey-level thresholding methods based on histogram or other statistic information about the interest image such as maximum entropy and so on have been proposed in the past. However, most methods based on statistic analysis of the images concerned little about the characteristics of morphology of interest objects, which sometimes could provide very important indication which can help to find the optimum threshold, especially for those organisms which have special texture morphologies such as vasculature, neuro-network etc. in medical imaging. In this paper, we propose a novel method for thresholding the fluorescent vasculature image series recorded from Confocal Scanning Laser Microscope. After extracting the basic orientation of the slice of vessels inside a sub-region partitioned from the images, we analysis the intensity profiles perpendicular to the vessel orientation to get the reasonable initial threshold for each region. Then the threshold values of those regions near the interest one both in x-y and optical directions have been referenced to get the final result of thresholds of the region, which makes the whole stack of images look more continuous. The resulting images are characterized by suppressing both noise and non-interest tissues conglutinated to vessels, while improving the vessel connectivities and edge definitions. The value of the method for idealized thresholding the fluorescence images of biological objects is demonstrated by a comparison of the results of 3D vascular reconstruction.

  11. Effects of photodynamic therapy on Gram-positive and Gram-negative bacterial biofilms by bioluminescence imaging and scanning electron microscopic analysis.

    PubMed

    Garcez, Aguinaldo S; Núñez, Silvia C; Azambuja, Nilton; Fregnani, Eduardo R; Rodriguez, Helena M H; Hamblin, Michael R; Suzuki, Hideo; Ribeiro, Martha S

    2013-11-01

    The aim of this study was to test photodynamic therapy (PDT) as an alternative approach to biofilm disruption on dental hard tissue, We evaluated the effect of methylene blue and a 660 nm diode laser on the viability and architecture of Gram-positive and Gram-negative bacterial biofilms. Ten human teeth were inoculated with bioluminescent Pseudomonas aeruginosa or Enterococcus faecalis to form 3 day biofilms in prepared root canals. Bioluminescence imaging was used to serially quantify and evaluate the bacterial viability, and scanning electron microscopic (SEM) imaging was used to assess architecture and morphology of bacterial biofilm before and after PDT employing methylene blue and 40 mW, 660 nm diode laser light delivered into the root canal via a 300 μm fiber for 240 sec, resulting in a total energy of 9.6 J. The data were statistically analyzed with analysis of variance (ANOVA) followed by Tukey test. The bacterial reduction showed a dose dependence; as the light energy increased, the bioluminescence decreased in both planktonic suspension and in biofilms. The SEM analysis showed a significant reduction of biofilm on the surface. PDT promoted disruption of the biofilm and the number of adherent bacteria was reduced. The photodynamic effect seems to disrupt the biofilm by acting both on bacterial cells and on the extracellular matrix.

  12. Image-guided feedback for ophthalmic microsurgery using multimodal intraoperative swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Li, Jianwei D.; Malone, Joseph D.; El-Haddad, Mohamed T.; Arquitola, Amber M.; Joos, Karen M.; Patel, Shriji N.; Tao, Yuankai K.

    2017-02-01

    Surgical interventions for ocular diseases involve manipulations of semi-transparent structures in the eye, but limited visualization of these tissue layers remains a critical barrier to developing novel surgical techniques and improving clinical outcomes. We addressed limitations in image-guided ophthalmic microsurgery by using microscope-integrated multimodal intraoperative swept-source spectrally encoded scanning laser ophthalmoscopy and optical coherence tomography (iSS-SESLO-OCT). We previously demonstrated in vivo human ophthalmic imaging using SS-SESLO-OCT, which enabled simultaneous acquisition of en face SESLO images with every OCT cross-section. Here, we integrated our new 400 kHz iSS-SESLO-OCT, which used a buffered Axsun 1060 nm swept-source, with a surgical microscope and TrueVision stereoscopic viewing system to provide image-based feedback. In vivo human imaging performance was demonstrated on a healthy volunteer, and simulated surgical maneuvers were performed in ex vivo porcine eyes. Denselysampled static volumes and volumes subsampled at 10 volumes-per-second were used to visualize tissue deformations and surgical dynamics during corneal sweeps, compressions, and dissections, and retinal sweeps, compressions, and elevations. En face SESLO images enabled orientation and co-registration with the widefield surgical microscope view while OCT imaging enabled depth-resolved visualization of surgical instrument positions relative to anatomic structures-of-interest. TrueVision heads-up display allowed for side-by-side viewing of the surgical field with SESLO and OCT previews for real-time feedback, and we demonstrated novel integrated segmentation overlays for augmented-reality surgical guidance. Integration of these complementary imaging modalities may benefit surgical outcomes by enabling real-time intraoperative visualization of surgical plans, instrument positions, tissue deformations, and image-based surrogate biomarkers correlated with completion of surgical goals.

  13. Evaluation of the Effects of Er,Cr:YSGG Laser, Ultrasonic Scaler and Curette on Root Surface Profile Using Surface Analyser and Scanning Electron Microscope: An In Vitro Study.

    PubMed

    Arora, Shipra; Lamba, Arundeep Kaur; Faraz, Farrukh; Tandon, Shruti; Ahad, Abdul

    2016-01-01

    Introduction: The periodontal therapy is primarily targeted at removal of dental plaque and plaque retentive factors. Although the thorough removal of adherent plaque, calculus and infected root cementum is desirable, it is not always achieved by conventional modalities. To accomplish more efficient results several alternative devices have been used. Lasers are one of the most promising modalities for nonsurgical periodontal treatment as they can achieve excellent tissue ablation with strong bactericidal and detoxification effects. Methods: Thirty freshly extracted premolars were selected and decoronated. The mesial surface of each root was divided vertically into four approximately equal parts. These were distributed into four group based on the root surface treatment. Part A (n = 30) was taken as control and no instrumentation was performed. Part B (n = 30) was irradiated by Erbium, Chromium doped Yttrium Scandium Gallium Garnet (Er,Cr:YSGG) laser. Part C (n = 30) was treated by piezoelectric ultrasonic scaler. Part D (n = 30) was treated by Gracey curette. The surface roughness was quantitatively analyzed by profilometer using roughness average (Ra) value, while presence of smear layer, cracks, craters and melting of surface were analyzed using scanning electron microscope (SEM). The means across the groups were statistically compared with control using Dunnett test. Results: Among the test groups, Er,Cr:YSGG laser group showed maximum surface roughness (mean Ra value of 4.14 μm) as compared to ultrasonic scaler (1.727 μm) and curette group (1.22 μm). However, surface with smear layer were found to be maximum (50%) in curette treated samples and minimum (20%) in laser treated ones. Maximum cracks (83.34%) were produced by ultrasonic scaler, and minimum (43.33%) by curettes. Crater formation was maximum (50%) in laser treated samples and minimum (3.33%) in curette treated ones. 63.33% samples treated by laser demonstrated melting of root surface, followed by ultrasonic scaler and curettes. Conclusion: Er,Cr:YSGG laser produced maximum microstructural changes on root surface that can influence the attachment of soft periodontal tissues as well as plaque and calculus deposition. In vivo studies are needed to validate these results and to evaluate their clinical effects.

  14. Surface nanotexturing of tantalum by laser ablation in water

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

    Barmina, E V; Simakin, Aleksandr V; Shafeev, Georgii A

    2009-01-31

    Surface nanotexturing of tantalum by ablation with short laser pulses in water has been studied experimentally using three ablation sources: a neodymium laser with a pulse duration of 350 ps, an excimer laser (248 nm) with a pulse duration of 5 ps and a Ti:sapphire laser with a pulse duration of 180 fs. The morphology of the nanotextured surfaces has been examined using a nanoprofilometer and field emission scanning electron microscope. The results demonstrate that the average size of the hillocks produced on the target surface depends on the laser energy density and is {approx}200 nm at an energy densitymore » approaching the laser-melting threshold of tantalum and a pulse duration of 350 ps. Their surface density reaches 10{sup 6} cm{sup -2}. At a pulse duration of 5 ps, the average hillock size is 60-70 nm. Nanotexturing is accompanied by changes in the absorption spectrum of the tantalum surface in the UV and visible spectral regions. The possible mechanisms of surface nanotexturing and potential applications of this effect are discussed. (nanostructures)« less

  15. Laser polishing of additive manufactured Ti alloys

    NASA Astrophysics Data System (ADS)

    Ma, C. P.; Guan, Y. C.; Zhou, W.

    2017-06-01

    Laser-based additive manufacturing has attracted much attention as a promising 3D printing method for metallic components in recent years. However, surface roughness of additive manufactured components has been considered as a challenge to achieve high performance. In this work, we demonstrate the capability of fiber laser in polishing rough surface of additive manufactured Ti-based alloys as Ti-6Al-4V and TC11. Both as-received surface and laser-polished surfaces as well as cross-section subsurfaces were analyzed carefully by White-Light Interference, Confocal Microscope, Focus Ion Beam, Scanning Electron Microscopy, Energy Dispersive Spectrometer, and X-ray Diffraction. Results revealed that as-received Ti-based alloys with surface roughness more than 5 μm could be reduce to less than 1 μm through laser polishing process. Moreover, microstructure, microhardness and wear resistance of laser-polished zone was investigated in order to examine the thermal effect of laser polishing processing on the substrate of additive manufactured Ti alloys. This proof-of-concept process has the potential to effectively improve the surface roughness of additive manufactured metallic alloy by local polishing method without damage to the substrate.

  16. Microscopic observation of laser glazed yttria-stabilized zirconia coatings

    NASA Astrophysics Data System (ADS)

    Morks, M. F.; Berndt, C. C.; Durandet, Y.; Brandt, M.; Wang, J.

    2010-08-01

    Thermal barrier coatings (TBCs) are frequently used as insulation system for hot components in gas-turbine, combustors and power plant industries. The corrosive gases which come from combustion of low grade fuels can penetrate into the TBCs and reach the metallic components and bond coat and cause hot corrosion and erosion damage. Glazing the top coat by laser beam is advanced approach to seal TBCs surface. The laser beam has the advantage of forming a dense thin layer composed of micrograins. Plasma-sprayed yttria-stabilized zirconia (YSZ) coating was glazed with Nd-YAG laser at different operating conditions. The surface morphologies, before and after laser treatment, were investigated by scanning electron microscopy. Laser beam assisted the densification of the surface by remelting a thin layer of the exposed surface. The laser glazing converted the rough surface of TBCs into smooth micron-size grains with size of 2-9 μm and narrow grain boundaries. The glazed surfaces showed higher Vickers hardness compared to as-sprayed coatings. The results revealed that the hardness increases as the grain size decreases.

  17. Modern technologies for retinal scanning and imaging: an introduction for the biomedical engineer

    PubMed Central

    2014-01-01

    This review article is meant to help biomedical engineers and nonphysical scientists better understand the principles of, and the main trends in modern scanning and imaging modalities used in ophthalmology. It is intended to ease the communication between physicists, medical doctors and engineers, and hopefully encourage “classical” biomedical engineers to generate new ideas and to initiate projects in an area which has traditionally been dominated by optical physics. Most of the methods involved are applicable to other areas of biomedical optics and optoelectronics, such as microscopic imaging, spectroscopy, spectral imaging, opto-acoustic tomography, fluorescence imaging etc., all of which are with potential biomedical application. Although all described methods are novel and important, the emphasis of this review has been placed on three technologies introduced in the 1990’s and still undergoing vigorous development: Confocal Scanning Laser Ophthalmoscopy, Optical Coherence Tomography, and polarization-sensitive retinal scanning. PMID:24779618

  18. Microstructure and properties of laser-clad high-temperature wear-resistant alloys

    NASA Astrophysics Data System (ADS)

    Yang, Yongqiang

    1999-02-01

    A 2-kW CO 2 laser with a powder feeder was used to produce alloy coatings with high temperature-wear resistance on the surface of steel substrates. To analyze the microstructure and microchemical composition of the laser-clad layers, a scanning electron microscope (SEM) equipped with an energy dispersive X-ray microanalysis system was employed. X-ray diffraction techniques were applied to characterize the phases formed during the cladding process. The results show that the microstructure of the cladding alloy consists mainly of many dispersed particles (W 2C, (W,Ti)C 1- x, WC), a lamellar eutectic carbide M 12C, and an (f.c.c) matrix. Hardness tested at room and high temperature showed that the laser-clad zone has a moderate room temperature hardness and relatively higher elevated temperature hardness. The application of the laser-clad layer to a hot tool was very successful, and its operational life span was prolonged 1 to 4 times.

  19. a Study on Microstructure Characteristics of IN SITU Formed TiC Reinforced Composite Coatings

    NASA Astrophysics Data System (ADS)

    Liu, Peng; Guo, Wei; Luo, Hui

    2012-04-01

    In situ synthesized TiC reinforced composite coating was fabricated by laser cladding of Al-Ni-Cr-C powders on titanium alloys, which can greatly improve the surface performance of the substrate. In this study, the Al-Ni-Cr-C laser-cladded composite coatings have been researched by means of X-ray diffraction, scanning electron microscope (SEM) and electron probe micro-analyzer (EPMA). There was a metallurgical combination between the Al-Ni-Cr-C laser-cladded coating and the Ti-6Al-4V substrate, and the micro-hardness of the Al-Ni-Cr-C laser-cladded coating was in the range of 1200-1450 HV0.2, which was 3-4 times higher than that of Ti-6Al-4V substrate. Furthermore, the reinforcement of theAl-Ni-Cr-C laser-cladded coating were mainly contributed to the action of the TiC, Ti3Al, Cr7C3, Al8Cr5 phases and the solution strengthening.

  20. Characterization of laser induced damage of HR coatings with picosecond pulses

    NASA Astrophysics Data System (ADS)

    Li, Cheng; Zhao, Yuan'an; Cui, Yun; Wang, Yueliang; Peng, Xiaocong; Shan, Chong; Zhu, Meiping; Wang, Jianguo; Shao, Jianda

    2017-11-01

    The effect of protective layer on the picosecond laser-induced damage behaviors of HfO2/SiO2 high-reflective (HR) coatings are explored. Two kinds of 1064nm HR coatings with and without protective layer are deposited by electron beam evaporation. Laser-induced damage tests are conducted with 1064nm, 30ps S-polarized and P-polarized pulses with different angle of incidence (AOI) to make the electric fields intensity in the HR coatings discrepantly. Damage morphology and cross section of damage sites were characterized by scanning electron microscope (SEM) and focused ion beam (FIB), respectively. It is found that SiO2 protective layer have a certain degree of improvement on laser induced damage threshold (LIDT) for every AOIs. The onset damage initiated very near to the Max peak of e-field, after which forms ripple-like pits. The damage morphology presents as layer delamination at high fluence. The Laser damage resistance is correspond with the maximum E-intensity in the coating stacks.

  1. Ultrafast amorphization in Ge(10)Sb(2)Te(13) thin film induced by single femtosecond laser pulse.

    PubMed

    Konishi, Mitsutaka; Santo, Hisashi; Hongo, Yuki; Tajima, Kazuyuki; Hosoi, Masaharu; Saiki, Toshiharu

    2010-06-20

    We demonstrate amorphization in a Ge(10)Sb(2)Te(13) (GST) thin film through a nonthermal process by femtosecond electronic excitation. Amorphous recording marks were formed by irradiation with a single femtosecond pulse, and were confirmed to be recrystallized by laser thermal annealing. Scanning electron microscope observations revealed that amorphization occurred below the melting temperature. We performed femtosecond pump-probe measurements to investigate the amorphization dynamics of a GST thin film. We found that the reflectivity dropped abruptly within 500fs after excitation by a single pulse and that a small change in the reflectivity occurred within 5ps of this drop.

  2. HA/Bioglass composite films deposited by pulsed laser with different substrate temperature

    NASA Astrophysics Data System (ADS)

    Wang, D. G.; Chen, C. Z.; Jin, Q. P.; Li, H. C.; Pan, Y. K.

    2014-03-01

    In this experiment, the HA/Bioglass composite films on Ti-6Al-4V were deposited by a pulsed laser at Ar atmosphere, and the influence of substrate temperature on the morphology, phase constitutions, bonding configurations and adhesive strength of the films was studied. The obtained films were characterized by an electron probe microanalyzer (EPMA), scanning electron microscope (SEM), X-ray diffractometer (XRD), Fourier transform infrared spectrometer (FTIR), scratch apparatus, and so on. The results show that the amount of the droplets, the crystallinity, and the critical load of the deposited films all increase with the increase of the substrate temperature; however, the substrate temperature has little influence on the functional groups of the films.

  3. Confocal laser endomicroscopy in the "in vivo" histological diagnosis of the gastrointestinal tract.

    PubMed

    De Palma, Giovanni D

    2009-12-14

    Recent technological advances in miniaturization have allowed for a confocal scanning microscope to be integrated into a conventional flexible endoscope, or into trans-endoscopic probes, a technique now known as confocal endomicroscopy or confocal laser endomicroscopy. This newly-developed technology has enabled endoscopists to collect real-time in vivo histological images or "virtual biopsies" of the gastrointestinal mucosa during endoscopy, and has stimulated significant interest in the application of this technique in clinical gastroenterology. This review aims to evaluate the current data on the technical aspects and the utility of this new technology in clinical gastroenterology and its potential impact in the future, particularly in the screening or surveillance of gastrointestinal neoplasia.

  4. TiCl4 as a source of TiO2 particles for laser anemometry measurements in hot gas

    NASA Technical Reports Server (NTRS)

    Weikle, Donald H.; Seasholtz, Richard G.; Oberle, Lawrence G.

    1990-01-01

    A method of reacting TiCl4 with water saturated gaseous nitrogen (GN2) at the entrance into a high temperature gas flow is described. The TiO2 particles formed are then entrained in the gas flow and used as seed particles for making laser anemometry (LA) measurements of the flow velocity distribution in the hot gas. Scanning electron microscope photographs of the TiO2 particles are shown. Data rate of the LA processor was measured to determine the amount of TiO2 formed. The TiCl4 and mixing gas flow diagram is shown. This work was performed in an open jet burner.

  5. Simple turbulence measurements with azopolymer thin films.

    PubMed

    Barillé, Regis; Pérez, Darío G; Morille, Yohann; Zielińska, Sonia; Ortyl, Ewelina

    2013-04-01

    A simple method to measure the influence on the laser beam propagation by a turbid medium is proposed. This measurement is based on the inscription of a surface relief grating (SRG) on an azopolymer thin film. The grating obtained with a single laser beam after propagation into a turbulent medium is perturbed and directly analyzed by a CCD camera through its diffraction pattern. Later, by scanning the surface pattern with an atomic force microscope, the inscribed SRG is analyzed with the Radon transform. This method has the advantage of using a single beam to remotely inscribe a grating detecting perturbations during the beam path. A method to evaluate the refractive index constant structure is developed.

  6. Comparative study of image contrast in scanning electron microscope and helium ion microscope.

    PubMed

    O'Connell, R; Chen, Y; Zhang, H; Zhou, Y; Fox, D; Maguire, P; Wang, J J; Rodenburg, C

    2017-12-01

    Images of Ga + -implanted amorphous silicon layers in a 110 n-type silicon substrate have been collected by a range of detectors in a scanning electron microscope and a helium ion microscope. The effects of the implantation dose and imaging parameters (beam energy, dwell time, etc.) on the image contrast were investigated. We demonstrate a similar relationship for both the helium ion microscope Everhart-Thornley and scanning electron microscope Inlens detectors between the contrast of the images and the Ga + density and imaging parameters. These results also show that dynamic charging effects have a significant impact on the quantification of the helium ion microscope and scanning electron microscope contrast. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  7. Image-guided removal of interproximal lesions with a CO2 laser

    NASA Astrophysics Data System (ADS)

    Ngo, Albert; Chan, Kenneth H.; Le, Oanh; Simon, Jacob C.; Fried, Daniel

    2018-02-01

    Recent studies have shown that near-IR (NIR) imaging methods such as NIR reflectance can be used to image lesions on proximal surfaces, and optical coherence tomography (OCT) can be used to measure the depth of those lesions below the tooth surface. These imaging modalities can be used to acquire high contrast images of demineralized tooth surfaces, and 2-D and 3-D images can be extracted from this data. At NIR wavelengths longer than 1200-nm, there is no interference from stains and the contrast is only due to the increased light scattering of the demineralization. Previous studies have shown that image-guided laser ablation can be used to remove occlusal lesions, but its use for the removal of subsurface lesions on proximal surfaces has not been investigated. The objective of this study is to demonstrate that simultaneously scanned NIR and CO2 lasers can be used to selectively remove natural and artificial interproximal caries lesions with minimal damage to sound tooth structure. In this study, images of simulated and natural interproximal lesions on extracted teeth were imaged using a digital microscope, a scanned 1460-nm superluminescent laser diode with an InGaAs detector and a cross polarization OCT system operating at 1300-nm. The lesions were subsequently removed with a CO2 laser operating at 9.3-μm and the dental handpiece and the volume of sound tissue removed was compared.

  8. Optical detection of ultrasound using an apertureless near-field scanning optical microscopy system

    NASA Astrophysics Data System (ADS)

    Ahn, Phillip; Zhang, Zhen; Sun, Cheng; Balogun, Oluwaseyi

    2013-01-01

    Laser ultrasonics techniques are power approaches for non-contact generation and detection of high frequency ultrasound on a local scale. In these techniques, optical diffraction limits the spatial information that can be accessed from a measurement. In order to improve the lateral spatial resolution, we incorporate an apertureless near-field scanning optical microscope (aNSOM) into laser ultrasonics setup for local detection of laser generated ultrasound. The aNSOM technique relies on the measurement of a weak backscattered near-field light intensity resulting from the oblique illumination of a nanoscale probe-tip positioned close to a sample surface. We enhance the optical near-field intensity by coupling light to surface plasmon polaritons (SPPs) on the shaft of an atomic force microscopy (AFM) cantilever. The SPPs propagate down the AFM shaft, localize at the tip apex, and are backscattered to the far-field when the separation distance between the probe tip and the sample surface is comparable to the probe-tip radius. The backscattered near-field intensity is dynamically modulated when an ultrasonic wave arrives at the sample surface leading to a transient change in the tip-sample separation distance. We present experimental results detailing measurement of broadband and narrowband laser generated ultrasound in solids with frequencies reaching up to 180 MHz range.

  9. A Student-Built Scanning Tunneling Microscope

    ERIC Educational Resources Information Center

    Ekkens, Tom

    2015-01-01

    Many introductory and nanotechnology textbooks discuss the operation of various microscopes including atomic force (AFM), scanning tunneling (STM), and scanning electron microscopes (SEM). In a nanotechnology laboratory class, students frequently utilize microscopes to obtain data without a thought about the detailed operation of the tool itself.…

  10. Three-dimensional behavior of ice crystals and biological cells during freezing of cell suspensions.

    PubMed

    Ishiguro, H; Koike, K

    1998-09-11

    Behavior of ice crystals and human red blood cells during extracellular-freezing was investigated in three-dimensions using a confocal laser scanning microscope(CLSM), which noninvasively produces tomograms of biological materials. Physiological saline and physiological saline with 2.4 M glycerol were used for suspension. Various cooling rates for directional solidification were used for distinctive morphology of the ice crystals. Addition of acridine orange as a fluorescent dye into the cell suspension enabled ice crystal, cells and unfrozen solution to be distinguished by different colors. The results indicate that the microscopic structure is three-dimensional for flat, cellular, and dendritic solid-liquid interfaces and that a CLSM is very effective in studying three-dimensional structure during the freezing of cell suspensions.

  11. Laser interferometry force-feedback sensor for an interfacial force microscope

    DOEpatents

    Houston, Jack E.; Smith, William L.

    2004-04-13

    A scanning force microscope is provided with a force-feedback sensor to increase sensitivity and stability in determining interfacial forces between a probe and a sample. The sensor utilizes an interferometry technique that uses a collimated light beam directed onto a deflecting member, comprising a common plate suspended above capacitor electrodes situated on a substrate forming an interference cavity with a probe on the side of the common plate opposite the side suspended above capacitor electrodes. The probe interacts with the surface of the sample and the intensity of the reflected beam is measured and used to determine the change in displacement of the probe to the sample and to control the probe distance relative to the surface of the sample.

  12. Analysis of protein and lipid dynamics using confocal fluorescence recovery after photobleaching (FRAP)

    PubMed Central

    Day, Charles A.; Kraft, Lewis J.; Kang, Minchul; Kenworthy, Anne K.

    2012-01-01

    Fluorescence recovery after photobleaching (FRAP) is a powerful, versatile and widely accessible tool to monitor molecular dynamics in living cells that can be performed using modern confocal microscopes. Although the basic principles of FRAP are simple, quantitative FRAP analysis requires careful experimental design, data collection and analysis. In this review we discuss the theoretical basis for confocal FRAP, followed by step-by-step protocols for FRAP data acquisition using a laser scanning confocal microscope for (1) measuring the diffusion of a membrane protein, (2) measuring the diffusion of a soluble protein, and (3) analysis of intracellular trafficking. Finally, data analysis procedures are discussed and an equation for determining the diffusion coefficient of a molecular species undergoing pure diffusion is presented. PMID:23042527

  13. A cell-free assay to determine the stoichiometry of plasma membrane proteins.

    PubMed

    Trigo, Cesar; Vivar, Juan P; Gonzalez, Carlos B; Brauchi, Sebastian

    2013-04-01

    Plasma membrane receptors, transporters, and ion channel molecules are often found as oligomeric structures that participate in signaling cascades essential for cell survival. Different states of protein oligomerization may play a role in functional control and allosteric regulation. Stochastic GFP-photobleaching (SGP) has emerged as an affordable and simple method to determine the stoichiometry of proteins at the plasma membrane. This non-invasive optical approach can be useful for total internal reflection of fluorescence microscopy (TIRFM), where signal-to-noise ratio is very high at the plasma membrane. Here, we report an alternative methodology implemented on a standard laser scanning confocal microscope (LSCM). The simplicity of our method will allow for its implementation in any epifluorescence microscope of choice.

  14. Atmospheric plasma surface modifications of electrospun PCL/chitosan/PCL hybrid scaffolds by nozzle type plasma jets for usage of cell cultivation

    NASA Astrophysics Data System (ADS)

    Surucu, Seda; Masur, Kai; Turkoglu Sasmazel, Hilal; Von Woedtke, Thomas; Weltmann, Klaus Dieter

    2016-11-01

    This paper reports Ar gas, Ar + O2, Ar + O2 + N2 gas mixtures and dry air plasma modifications by atmospheric pressure argon driven kINPen and air driven Diener (PlasmaBeam) plasma jets to alter surface properties of three dimensional (3D), electrospun PCL/Chitosan/PCL layer by layer hybrid scaffolds to improve human fibroblast (MRC5) cell attachment and growth. The characterizations of the samples were done by contact angle (CA) measurements, scanning electron microscopy (SEM), X-Ray Photoelectron spectroscopy (XPS) analysis. The results showed that the plasma modification carried out under dry air and Ar + O2 + N2 gas mixtures were altered effectively the nanotopography and the functionality of the material surfaces. It was found that the samples treated with Ar + O2 + N2 gas mixtures for 1 min and dry air for 9 min have better hydrophilicity 78.9° ± 1.0 and 75.6° ± 0.1, respectively compared to the untreated samples (126.5°). Biocompatibility performance of the scaffolds was determined with alamarBlue (aB) assay and MTT assay methods, Giemsa staining, fluorescence microscope, confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) analyses. The results showed that plasma treated samples increased the hydrophilicity and oxygen functionality and topography of the surfaces significantly, thus affecting the cell viability and proliferation on/within scaffolds.

  15. Efficacy of Sodium Hypochlorite Activated With Laser in Intracanal Smear Layer Removal: An SEM Study

    PubMed Central

    Shahriari, Shahriar; Kasraei, Shahin; Roshanaei, Ghodratollah; Karkeabadi, Hamed; Davanloo, Hossein

    2017-01-01

    Introduction: The purpose of the present study was to evaluate the different concentrations of sodium hypochlorite activated with laser in removing of the smear layer in the apical, middle, and coronal segments of root canal walls by scanning electron microscopy analysis. Methods: Sixty single-rooted human mandibular teeth were decoronated to a standardized length. The samples were prepared by using Race rotary system to size 40, 0.04 taper and divided into 4 equal groups (n = 15). Group 1, irrigated with EDTA 17% and 5.25% NaOCl, groups 2, 3 and 4, 1%, 2.5%, and 5% NaOCl activated with Nd:YAG laser, respectively. Teeth were split longitudinally and subjected to scanning electron microscope (SEM). Data were analyzed by Kruskal-Wallis, Mann-Whitney tests. P value of <0.05 was considered statistically significant. Results: Five percent NaOCl LAI (laser-activated irrigation) showed best smear layer removal in test groups and the difference was statistically significant (P < 0.001). Control group (EDTA 17% and 5.25% NaOCl irrigation) showed significantly better outcomes in comparative with test groups (P < 0.001). In the apical third, compared to coronal and middle third, the canal walls were often contaminated by inorganic debris and smear layer. Conclusion: All different concentrations of sodium hypochlorite activated with laser have a positive effect on removing of smear layer. Sodium hypochlorite activated with laser removed smear layer more effectively at the coronal and middle third compared to the apical third. PMID:28912942

  16. Modification of tribology and high-temperature behavior of Ti 48Al 2Cr 2Nb intermetallic alloy by laser cladding

    NASA Astrophysics Data System (ADS)

    Liu, Xiu-Bo; Wang, Hua-Ming

    2006-06-01

    In order to improve the tribology and high-temperature oxidation properties of the Ti-48Al-2Cr-2Nb intermetallic alloy simultaneously, mixed NiCr-Cr 3C 2 precursor powders had been investigated for laser cladding treatment to modify wear and high-temperature oxidation resistance of the material. The alloy samples were pre-placed with NiCr-80, 50 and 20%Cr 3C 2 (wt.%), respectively, and laser treated at the same parameters, i.e., laser output power 2.8 kW, beam scanning speed 2.0 mm/s, beam dimension 1 mm × 18 mm. The treated samples underwent tests of microhardness, wear and high-temperature oxidation. The results showed that laser cladding with different constitution of mixed precursor NiCr-Cr 3C 2 powders improved surface hardness in all cases. Laser cladding with NiCr-50%Cr 3C 2 resulted in the best modification of tribology and high-temperature oxidation behavior. X-ray diffraction (XRD), optical microscope (OM), scanning electron microscopy (SEM) and energy-dispersive spectrometer (EDS) analyses indicated that the formation of reinforced Cr 7C 3, TiC and both continuous and dense Al 2O 3, Cr 2O 3 oxide scales were supposed to be responsible for the modification of the relevant properties. As a result, the present work had laid beneficial surface engineering foundation for TiAl alloy applied as future light weight and high-temperature structural candidate materials.

  17. Surface imaging microscope

    NASA Astrophysics Data System (ADS)

    Rogala, Eric W.; Bankman, Isaac N.

    2008-04-01

    The three-dimensional shapes of microscopic objects are becoming increasingly important for battlespace CBRNE sensing. Potential applications of microscopic 3D shape observations include characterization of biological weapon particles and manufacturing of micromechanical components. Aerosol signatures of stand-off lidar systems, using elastic backscatter or polarization, are dictated by the aerosol particle shapes and sizes that must be well characterized in the lab. A low-cost, fast instrument for 3D surface shape microscopy will be a valuable point sensor for biological particle sensing applications. Both the cost and imaging durations of traditional techniques such as confocal microscopes, atomic force microscopes, and electron scanning microscopes are too high. We investigated the feasibility of a low-cost, fast interferometric technique for imaging the 3D surface shape of microscopic objects at frame rates limited only by the camera in the system. The system operates at two laser wavelengths producing two fringe images collected simultaneously by a digital camera, and a specialized algorithm we developed reconstructs the surface map of the microscopic object. The current implementation assembled to test the concept and develop the new 3D reconstruction algorithm has 0.25 micron resolution in the x and y directions, and about 0.1 micron accuracy in the z direction, as tested on a microscopic glass test object manufactured with etching techniques. We describe the interferometric instrument, present the reconstruction algorithm, and discuss further development.

  18. Confocal laser scanning microscopy and area-scale analysis used to quantify enamel surface textural changes from citric acid demineralization and salivary remineralization in vitro.

    PubMed

    Austin, R S; Giusca, C L; Macaulay, G; Moazzez, R; Bartlett, D W

    2016-02-01

    This paper investigates the application of confocal laser scanning microscopy to determine the effect of acid-mediated erosive enamel wear on the micro-texture of polished human enamel in vitro. Twenty polished enamel samples were prepared and subjected to a citric acid erosion and pooled human saliva remineralization model. Enamel surface microhardness was measured using a Knoop hardness tester, which confirmed that an early enamel erosion lesion was formed which was then subsequently completely remineralized. A confocal laser scanning microscope was used to capture high-resolution images of the enamel surfaces undergoing demineralization and remineralization. Area-scale analysis was used to identify the optimal feature size following which the surface texture was determined using the 3D (areal) texture parameter Sa. The Sa successfully characterized the enamel erosion and remineralization for the polished enamel samples (P<0.001). Areal surface texture characterization of the surface events occurring during enamel demineralization and remineralization requires optical imaging instrumentation with lateral resolution <2.5 μm, applied in combination with appropriate filtering in order to remove unwanted waviness and roughness. These techniques will facilitate the development of novel methods for measuring early enamel erosion lesions in natural enamel surfaces in vivo. Copyright © 2015 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  19. Application of Laser Scanning Confocal Microscopy to Heat and Mass Transport Modeling in Porous Microstructures

    NASA Technical Reports Server (NTRS)

    Marshall, Jochen; Milos, Frank; Fredrich, Joanne; Rasky, Daniel J. (Technical Monitor)

    1997-01-01

    Laser Scanning Confocal Microscopy (LSCM) has been used to obtain digital images of the complicated 3-D (three-dimensional) microstructures of rigid, fibrous thermal protection system (TPS) materials. These orthotropic materials are comprised of refractory ceramic fibers with diameters in the range of 1 to 10 microns and have open porosities of 0.8 or more. Algorithms are being constructed to extract quantitative microstructural information from the digital data so that it may be applied to specific heat and mass transport modeling efforts; such information includes, for example, the solid and pore volume fractions, the internal surface area per volume, fiber diameter distributions, and fiber orientation distributions. This type of information is difficult to obtain in general, yet it is directly relevant to many computational efforts which seek to model macroscopic thermophysical phenomena in terms of microscopic mechanisms or interactions. Two such computational efforts for fibrous TPS materials are: i) the calculation of radiative transport properties; ii) the modeling of gas permeabilities.

  20. Tracking features in retinal images of adaptive optics confocal scanning laser ophthalmoscope using KLT-SIFT algorithm

    PubMed Central

    Li, Hao; Lu, Jing; Shi, Guohua; Zhang, Yudong

    2010-01-01

    With the use of adaptive optics (AO), high-resolution microscopic imaging of living human retina in the single cell level has been achieved. In an adaptive optics confocal scanning laser ophthalmoscope (AOSLO) system, with a small field size (about 1 degree, 280 μm), the motion of the eye severely affects the stabilization of the real-time video images and results in significant distortions of the retina images. In this paper, Scale-Invariant Feature Transform (SIFT) is used to abstract stable point features from the retina images. Kanade-Lucas-Tomasi(KLT) algorithm is applied to track the features. With the tracked features, the image distortion in each frame is removed by the second-order polynomial transformation, and 10 successive frames are co-added to enhance the image quality. Features of special interest in an image can also be selected manually and tracked by KLT. A point on a cone is selected manually, and the cone is tracked from frame to frame. PMID:21258443

  1. Nonlinear Focal Modulation Microscopy.

    PubMed

    Zhao, Guangyuan; Zheng, Cheng; Kuang, Cuifang; Zhou, Renjie; Kabir, Mohammad M; Toussaint, Kimani C; Wang, Wensheng; Xu, Liang; Li, Haifeng; Xiu, Peng; Liu, Xu

    2018-05-11

    We demonstrate nonlinear focal modulation microscopy (NFOMM) to achieve superresolution imaging. Traditional approaches to superresolution that utilize point scanning often rely on spatially reducing the size of the emission pattern by directly narrowing (e.g., through minimizing the detection pinhole in Airyscan, Zeiss) or indirectly peeling its outer profiles [e.g., through depleting the outer emission region in stimulated emission depletion (STED) microscopy]. We show that an alternative conceptualization that focuses on maximizing the optical system's frequency shifting ability offers advantages in further improving resolution while reducing system complexity. In NFOMM, a spatial light modulator and a suitably intense laser illumination are used to implement nonlinear focal-field modulation to achieve a transverse spatial resolution of ∼60  nm (∼λ/10). We show that NFOMM is comparable with STED microscopy and suitable for fundamental biology studies, as evidenced in imaging nuclear pore complexes, tubulin and vimentin in Vero cells. Since NFOMM is readily implemented as an add-on module to a laser-scanning microscope, we anticipate wide utility of this new imaging technique.

  2. [Preparation of panax notoginseng saponins-tanshinone H(A) composite method for pulmonary delivery with spray-drying method and its characterization].

    PubMed

    Wang, Hua-Mei; Fu, Ting-Ming; Guo, Li-Wei

    2013-02-01

    To prepare panax notoginseng saponins-tanshinone II(A) composite particles for pulmonary delivery, in order to explore a dry powder particle preparation method ensuring synchronized arrival of multiple components of traditional Chinese medicine compounds at absorption sites. Panax notoginseng saponins-tanshinone II(A) composite particles were prepared with spray-drying method, and characterized by scanning electron microscopy (SEM), confocal laser scanning microscope (CLSM), X-ray diffraction (XRD), infrared analysis (IR), dry laser particle size analysis, high performance liquid chromatography (HPLC) and the aerodynamic behavior was evaluated by a Next Generation Impactor (NGI). The dry powder particles produced had narrow particle size distribution range and good aerodynamic behavior, and could realize synchronized administration of multiple components. The spray-drying method is used to combine traditional Chinese medicine components with different physical and chemical properties in the same particle, and product into traditional Chinese medicine compound particles in line with the requirements for pulmonary delivery.

  3. Nonlinear Focal Modulation Microscopy

    NASA Astrophysics Data System (ADS)

    Zhao, Guangyuan; Zheng, Cheng; Kuang, Cuifang; Zhou, Renjie; Kabir, Mohammad M.; Toussaint, Kimani C.; Wang, Wensheng; Xu, Liang; Li, Haifeng; Xiu, Peng; Liu, Xu

    2018-05-01

    We demonstrate nonlinear focal modulation microscopy (NFOMM) to achieve superresolution imaging. Traditional approaches to superresolution that utilize point scanning often rely on spatially reducing the size of the emission pattern by directly narrowing (e.g., through minimizing the detection pinhole in Airyscan, Zeiss) or indirectly peeling its outer profiles [e.g., through depleting the outer emission region in stimulated emission depletion (STED) microscopy]. We show that an alternative conceptualization that focuses on maximizing the optical system's frequency shifting ability offers advantages in further improving resolution while reducing system complexity. In NFOMM, a spatial light modulator and a suitably intense laser illumination are used to implement nonlinear focal-field modulation to achieve a transverse spatial resolution of ˜60 nm (˜λ /10 ). We show that NFOMM is comparable with STED microscopy and suitable for fundamental biology studies, as evidenced in imaging nuclear pore complexes, tubulin and vimentin in Vero cells. Since NFOMM is readily implemented as an add-on module to a laser-scanning microscope, we anticipate wide utility of this new imaging technique.

  4. 3D imaging of cement-based materials at submicron resolution by combining laser scanning confocal microscopy with serial sectioning.

    PubMed

    Yio, M H N; Mac, M J; Wong, H S; Buenfeld, N R

    2015-05-01

    In this paper, we present a new method to reconstruct large volumes of nontransparent porous materials at submicron resolution. The proposed method combines fluorescence laser scanning confocal microscopy with serial sectioning to produce a series of overlapping confocal z-stacks, which are then aligned and stitched based on phase correlation. The method can be extended in the XY plane to further increase the overall image volume. Resolution of the reconstructed image volume does not degrade with increase in sample size. We have used the method to image cementitious materials, hardened cement paste and concrete and the results obtained show that the method is reliable. Possible applications of the method such as three-dimensional characterization of the pores and microcracks in hardened concrete, three-dimensional particle shape characterization of cementitious materials and three-dimensional characterization of other porous materials such as rocks and bioceramics are discussed. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  5. Two-photon excitation laser scanning microscopy of porcine nasal septal cartilage following Nd:YAG laser-mediated stress relaxation

    NASA Astrophysics Data System (ADS)

    Kim, Charlton C.; Wallace, Vincent P.; Rasouli, Alexandre; Coleno, Mariah L.; Dao, Xavier; Tromberg, Bruce J.; Wong, Brian J.

    2000-05-01

    Laser irradiation of hyaline cartilage result in stable shape changes due to temperature dependent stress relaxation. In this study, we determined the structural changes in chondrocytes within porcine nasal septal cartilage tissue over a 4-day period using a two-photon laser scanning microscope (TPM) following Nd:YAG laser irradiation (lambda equals 1.32 micrometer) using parameters that result in mechanical stress relaxation (6.0 W, 5.4 mm spot diameter). TPM excitation (780 nm) result in induction of fluorescence from endogenous agents such as NADH, NADPH, and flavoproteins in the 400 - 500 nm spectral region. During laser irradiation diffuse reflectance (from a probe HeNe laser, (lambda) equals 632.8 nm), surface temperature, and stress relaxation were measured dynamically. Each specimen received one, two, or three sequential laser exposures (average irradiation times of 5, 6, and 8 seconds). The cartilage reached a peak surface temperature of about 70 degrees Celsius during irradiation. Cartilage denatured in 50% EtOH (20 minutes) was used as a positive control. TPM was performed using a mode-locked 780 nm Titanium:Sapphire (Ti:Al203) beam with a, 63X, 1.2 N.A. water immersion objective (working distance of 200 mm) to detect the fluorescence emission from the chondrocytes. Images of chondrocytes were obtained at depths up to 150 microns (lateral resolution equals 35 micrometer X 35 micrometer). Images were obtained immediately following laser exposure, and also after 4 days in culture. In both cases, the irradiated and non-irradiated specimens do not show any discernible difference in general shape or auto fluorescence. In contrast, positive controls (immersed in 50% ethanol), show markedly increased fluorescence relative to both the native and irradiated specimens, in the cytoplasmic region.

  6. Excimer laser deinsulation of Parylene-C on iridium for use in an activated iridium oxide film-coated Utah electrode array

    PubMed Central

    Yoo, Je-Min; Negi, Sandeep; Tathireddy, Prashant; Solzbacher, Florian; Song, Jong-In; Rieth, Loren W.

    2013-01-01

    Implantable microelectrodes provide a measure to electrically stimulate neurons in the brain and spinal cord and record their electrophysiological activity. A material with a high charge capacity such as activated or sputter-deposited iridium oxide film (AIROF or SIROF) is used as an interface. The Utah electrode array (UEA) uses SIROF for its interface material with neural tissue and oxygen plasma etching (OPE) with an aluminium foil mask to expose the active area, where the interface between the electrode and neural tissue is formed. However, deinsulation of Parylene-C using OPE has limitations, including the lack of uniformity in the exposed area and reproducibility. While the deinsulation of Parylene-C using an excimer laser is proven to be an alternative for overcoming the limitations, the iridium oxide (IrOx) suffers from fracture when high laser fluence (>1000 mJ/cm2) is used. Iridium (Ir), which has a much higher fracture resistance than IrOx, can be deposited before excimer laser deinsulation and then the exposed Ir film area can be activated by electrochemical treatment to acquire the AIROF. Characterisation of the laser-ablated Ir film and AIROF by surface analysis (X-ray photoelectron spectroscopy, scanning electron microscope, and atomic force microscope) and electrochemical analysis (electrochemical impedance spectroscopy, and cyclic voltammetry) shows that the damage on the Ir film induced by laser irradiation is significantly less than that on SIROF, and the AIROF has a high charge storage capacity. The results show the potential of the laser deinsulation technique for use in high performance AIROF-coated UEA fabrication. PMID:23458659

  7. The oldest micropepline beetle from Cretaceous Burmese amber and its phylogenetic implications (Coleoptera: Staphylinidae)

    NASA Astrophysics Data System (ADS)

    Cai, Chen-Yang; Huang, Di-Ying

    2014-10-01

    The staphylinid subfamily Micropeplinae includes small strongly sclerotized beetles with truncate elytra leaving the most part of abdomen exposed. Fossil micropeplines are rare and confined to Cenozoic representatives of extant genera. Here, we describe the oldest micropepline, Protopeplus cretaceus gen. and sp. n., from the Upper Cretaceous Burmese amber. Fluorescence microscope and confocal laser scanning microscopy (CLSM) were both used to reveal diagnostic features of Micropeplinae and some primitive traits that place Protopeplus very basally within Micropeplinae.

  8. Generation of a frequency comb and applications thereof

    DOEpatents

    Hagmann, Mark J; Yarotski, Dmitry A

    2013-12-03

    Apparatus for generating a microwave frequency comb (MFC) in the DC tunneling current of a scanning tunneling microscope (STM) by fast optical rectification, cause by nonlinearity of the DC current vs. voltage curve for the tunneling junction, of regularly-spaced, short pulses of optical radiation from a focused mode-locked, ultrafast laser, directed onto the tunneling junction, is described. Application of the MFC to high resolution dopant profiling in semiconductors is simulated. Application of the MFC to other measurements is described.

  9. Acquisition of a Laser Scanning Confocal Microscope to Examine CNS Activity of Antidotal Oximes and to Enhance Undergraduate Research Training Across the Sciences

    DTIC Science & Technology

    2016-07-15

    Activity of Antidotal Oximes and to Enhance Undergraduate Research Training Across the Sciences The views, opinions and/or findings contained in this...to Examine CNS Activity of Antidotal Oximes and to Enhance Undergraduate Research Training Across the Sciences Report Title The project utilized...examining the ability of antidotal oximes to rescue organophosphate (OP)-induced CNS toxicity and training across the sciences and social sciences at

  10. Scanning fluorescent microscopy is an alternative for quantitative fluorescent cell analysis.

    PubMed

    Varga, Viktor Sebestyén; Bocsi, József; Sipos, Ferenc; Csendes, Gábor; Tulassay, Zsolt; Molnár, Béla

    2004-07-01

    Fluorescent measurements on cells are performed today with FCM and laser scanning cytometry. The scientific community dealing with quantitative cell analysis would benefit from the development of a new digital multichannel and virtual microscopy based scanning fluorescent microscopy technology and from its evaluation on routine standardized fluorescent beads and clinical specimens. We applied a commercial motorized fluorescent microscope system. The scanning was done at 20 x (0.5 NA) magnification, on three channels (Rhodamine, FITC, Hoechst). The SFM (scanning fluorescent microscopy) software included the following features: scanning area, exposure time, and channel definition, autofocused scanning, densitometric and morphometric cellular feature determination, gating on scatterplots and frequency histograms, and preparation of galleries of the gated cells. For the calibration and standardization Immuno-Brite beads were used. With application of shading compensation, the CV of fluorescence of the beads decreased from 24.3% to 3.9%. Standard JPEG image compression until 1:150 resulted in no significant change. The change of focus influenced the CV significantly only after +/-5 microm error. SFM is a valuable method for the evaluation of fluorescently labeled cells. Copyright 2004 Wiley-Liss, Inc.

  11. Surface changes of composite and compomer materials following irradiation with an Er:YAG laser and a frequency doubled alexandrite laser

    NASA Astrophysics Data System (ADS)

    Pilgrim, Christian G.; Rechmann, Peter; Hennig, Thomas; Goldin, Dan S.

    1999-05-01

    Er:YAG laser as well as the frequency doubled Alexandrite laser have been suggested for the use in periodontal therapy and so for the elimination of calculus and the treatment of infected root cement. Intended is the laser application inside the gingival pocket. In consequence, both these lasers may be used in areas close to cervical or approximal location, or even on the fillings' surfaces. Light cured composite and compomer materials are in use for these types of fillings among others. Aim of the study presented here was to compare the effect of an Er:YAG laser (wavelength 2.94 μm, pulse duration 250 μs, free running. fluence 4.5 J/cm2 and 21.4 J/cm2) on compomers and composites with the result following irradiation with a frequency doubled Alexandrite laser (wavelength 377 nm, pulse duration 200 ns, fluence 3 Jcm-2 and 6 J/cm2). The surface of standardized compomer and composite samples were irradiated with both laser wavelengths (either frequency doubled Alexandrite or Er:YAG laser) using the same standardized application protocol. Scanning electron microscopic investigation showed that irradiation with both lasers causes surface changes in composites and compomers removing calculus with these lasers.

  12. Electrophoretic Detection and Confocal Microscopic Imaging of Tyrosine Nitrated Proteins in Plant Tissue.

    PubMed

    Arora, Dhara; Singh, Neha; Bhatla, Satish C

    2018-01-01

    Tyrosine nitrated proteins can be detected in plant cells electrophoretically and their distribution can be monitored by confocal laser scanning microscopy (CLSM) imaging. One-dimensional polyacrylamide gel electrophoresis (1D PAGE) followed by Western blotting using polyclonal antibody against 3-nitrotyrosine residues enables detection of tyrosine nitrated proteins in plant cells. Here we describe detection of tyrosine nitrated proteins in the homogenates derived from sunflower (Helianthus annuus L.) seedling cotyledons. Total soluble proteins obtained from tissue homogenates are resolved using vertical gel electrophoresis followed by their electrophoretic transfer on to a microporous membrane support for immunodetection. Spatial distribution of tyrosine nitrated proteins can be visualized using an antibody against 3-nitrotyrosine residues. Immunofluorescent localization is performed by cutting 7 μm thick wax sections of tissue followed by incubation in primary anti-nitrotyrosine antibody (dilution 1:200) and secondary Cy-3 labeled anti-rabbit IgG antibody (dilution 1:1500). Confocal laser scanning microscopy analysis is undertaken using argon lasers (ex: 530-550 nm and em: 570 nm) at pinhole 1. Modulation in the abundance and spatial localization of tyrosine nitrated proteins in plant tissues can be monitored using these techniques.

  13. Motion-invariant SRT treatment detection from direct M-scan OCT imaging.

    PubMed

    Fountoukidou, Tatiana; Raisin, Philippe; Kaufmann, Daniel; Justiz, Jörn; Sznitman, Raphael; Wolf, Sebastian

    2018-03-08

    Selective retina therapy (SRT) is a laser treatment targeting specific posterior retinal layers. It is focused on inducing damage to the retinal pigment epithelium (RPE), while sparing other retinal tissue compared to traditional photocoagulation. However, the targeted RPE layer is invisible with most imaging modalities and induced SRT lesions cannot be monitored. In this work, imaging scans acquired from an experimental setup that couples the SRT laser beam with an optical coherence tomography (OCT) beam are analyzed in order to evaluate the treatment as they occur. We isolated a small part of the time-resolved scan corresponding to the end of the treatment, for which we have microscopic evidence of the SRT outcome. We then use a convolutional neural network to correspond each scan to the treatment result. We explore which aspects of the scan convey more valuable information for a robust therapy evaluation. By only using this adequately small part, we can achieve an online estimation, while being resilient to eye movement. The available dataset consists of time- resolved OCT scans of 98 ex vivo porcine eyes, treated with different energy levels. The proposed method yields high performance in the task of predicting whether the applied energy was adequate for SRT treatment, by focusing on the immediate OCT signal acquired during treatment time. We propose a strategy toward online noninvasive SRT treatment assessment, able to provide a satisfying evaluation of a treatment status, that therefore could be used for the planning of the treatment continuation.

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

    Ji, Yanfeng; Hui, Fei; Shi, Yuanyuan

    The conductive atomic force microscope (CAFM) has become an essential tool for the nanoscale electronic characterization of many materials and devices. When studying photoactive samples, the laser used by the CAFM to detect the deflection of the cantilever can generate photocurrents that perturb the current signals collected, leading to unreliable characterization. In metal-coated semiconductor samples, this problem is further aggravated, and large currents above the nanometer range can be observed even without the application of any bias. Here we present the first characterization of the photocurrents introduced by the laser of the CAFM, and we quantify the amount of lightmore » arriving to the surface of the sample. The mechanisms for current collection when placing the CAFM tip on metal-coated photoactive samples are also analyzed in-depth. Finally, we successfully avoided the laser-induced perturbations using a two pass technique: the first scan collects the topography (laser ON) and the second collects the current (laser OFF). We also demonstrate that CAFMs without a laser (using a tuning fork for detecting the deflection of the tip) do not have this problem.« less

  15. Laser cleaning of ITER's diagnostic mirrors

    NASA Astrophysics Data System (ADS)

    Skinner, C. H.; Gentile, C. A.; Doerner, R.

    2012-10-01

    Practical methods to clean ITER's diagnostic mirrors and restore reflectivity will be critical to ITER's plasma operations. We report on laser cleaning of single crystal molybdenum mirrors coated with either carbon or beryllium films 150 - 420 nm thick. A 1.06 μm Nd laser system provided 220 ns pulses at 8 kHz with typical power densities of 1-2 J/cm^2. The laser beam was fiber optically coupled to a scanner suitable for tokamak applications. The efficacy of mirror cleaning was assessed with a new technique that combines microscopic imaging and reflectivity measurements [1]. The method is suitable for hazardous materials such as beryllium as the mirrors remain sealed in a vacuum chamber. Excellent restoration of reflectivity for the carbon coated Mo mirrors was observed after laser scanning under vacuum conditions. For the beryllium coated mirrors restoration of reflectivity has so far been incomplete and modeling indicates that a shorter duration laser pulse is needed. No damage of the molybdenum mirror substrates was observed.[4pt][1] C.H. Skinner et al., Rev. Sci. Instrum. at press.

  16. Microstructure and wear resistance of laser cladded composite coatings prepared from pre-alloyed WC-NiCrMo powder with different laser spots

    NASA Astrophysics Data System (ADS)

    Yao, Jianhua; Zhang, Jie; Wu, Guolong; Wang, Liang; Zhang, Qunli; Liu, Rong

    2018-05-01

    The distribution of WC particles in laser cladded composite coatings can significantly affect the wear resistance of the coatings under aggressive environments. In this study, pre-alloyed WC-NiCrMo powder is deposited on SS316L via laser cladding with circular spot and wide-band spot, respectively. The microstructure and WC distribution of the coatings are investigated with optical microscope (OM), scanning electron microscopy (SEM), energy dispersive spectrometer (EDS), and X-ray diffraction (XRD). The wear behavior of the coatings is investigated under dry sliding-wear test. The experimental results show that the partially dissolved WC particles are uniformly distributed in both coatings produced with circular spot and wide-band spot, respectively, and the microstructures consist of WC and M23C6 carbides and γ-(Ni, Fe) solid solution matrix. However, due to Fe dilution, the two coatings have different microstructural characteristics, resulting in different hardness and wear resistance. The wide-band spot laser prepared coating shows better performance than the circular spot laser prepared coating.

  17. Corrosion resistance of a laser spot-welded joint of NiTi wire in simulated human body fluids.

    PubMed

    Yan, Xiao-Jun; Yang, Da-Zhi

    2006-04-01

    The purpose of this study was to investigate corrosion resistance of a laser spot-welded joint of NiTi alloy wires using potentiodynamic tests in Hank's solution at different PH values and the PH 7.4 NaCl solution for different Cl- concentrations. Scanning electron microscope observations were carried out before and after potentiodynamic tests. The composition of a laser spot-welded joint and base metal were characterized by using an electron probe microanalyzer. The results of potentiodynamic tests showed that corrosion resistance of a laser spot-welded joint of NiTi alloy wire was better than that of base metal, which exhibited a little higher breakdown potential and passive range, and a little lower passive current density. Corrosion resistances of a laser spot-welded joint and base metal decreased with increasing of the Cl- concentration and PH value. The improvement of corrosion resistance of the laser spot-welded joint was due to the decrease of the surface defects and the increase of the Ti/Ni ratio. (c) 2005 Wiley Periodicals, Inc.

  18. Microstructures and Properties of Laser Cladding Al-TiC-CeO2 Composite Coatings

    PubMed Central

    Kong, Dejun; Song, Renguo

    2018-01-01

    Al-TiC-CeO2 composite coatings have been prepared by using a laser cladding technique, and the microstructure and properties of the resulting composite coatings have been investigated using scanning electron microscopy (SEM), a 3D microscope system, X-ray diffraction (XRD), micro-hardness testing, X-ray stress measurements, friction and wear testing, and an electrochemical workstation. The results showed that an Al-Fe phase appears in the coatings under different applied laser powers and shows good metallurgical bonding with the matrix. The dilution rate of the coating first decreases and then increases with increasing laser power. The coating was transformed from massive and short rod-like structures into a fine granular structure, and the effect of fine grain strengthening is significant. The microhardness of the coatings first decreases and then increases with increasing laser power, and the maximum microhardness can reach 964.3 HV0.2. In addition, the residual stress of the coating surface was tensile stress, and crack size increases with increasing stress. When the laser power was 1.6 kW, the coating showed high corrosion resistance. PMID:29373555

  19. Microstructures and Properties of Laser Cladding Al-TiC-CeO₂ Composite Coatings.

    PubMed

    He, Xing; Kong, Dejun; Song, Renguo

    2018-01-26

    Al-TiC-CeO₂ composite coatings have been prepared by using a laser cladding technique, and the microstructure and properties of the resulting composite coatings have been investigated using scanning electron microscopy (SEM), a 3D microscope system, X-ray diffraction (XRD), micro-hardness testing, X-ray stress measurements, friction and wear testing, and an electrochemical workstation. The results showed that an Al-Fe phase appears in the coatings under different applied laser powers and shows good metallurgical bonding with the matrix. The dilution rate of the coating first decreases and then increases with increasing laser power. The coating was transformed from massive and short rod-like structures into a fine granular structure, and the effect of fine grain strengthening is significant. The microhardness of the coatings first decreases and then increases with increasing laser power, and the maximum microhardness can reach 964.3 HV 0.2 . In addition, the residual stress of the coating surface was tensile stress, and crack size increases with increasing stress. When the laser power was 1.6 kW, the coating showed high corrosion resistance.

  20. Spin microscope based on optically detected magnetic resonance

    DOEpatents

    Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

    2010-06-29

    The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

  1. Spin microscope based on optically detected magnetic resonance

    DOEpatents

    Berman, Gennady P.; Chernobrod, Boris M.

    2009-11-10

    The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of impaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

  2. Spin microscope based on optically detected magnetic resonance

    DOEpatents

    Berman, Gennady P.; Chernobrod, Boris M.

    2007-12-11

    The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

  3. Spin microscope based on optically detected magnetic resonance

    DOEpatents

    Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

    2010-07-13

    The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

  4. Spin microscope based on optically detected magnetic resonance

    DOEpatents

    Berman, Gennady P [Los Alamos, NM; Chernobrod, Boris M [Los Alamos, NM

    2009-10-27

    The invention relates to scanning magnetic microscope which has a photoluminescent nanoprobe implanted in the tip apex of an atomic force microscope (AFM), a scanning tunneling microscope (STM) or a near-field scanning optical microscope (NSOM) and exhibits optically detected magnetic resonance (ODMR) in the vicinity of unpaired electron spins or nuclear magnetic moments in the sample material. The described spin microscope has demonstrated nanoscale lateral resolution and single spin sensitivity for the AFM and STM embodiments.

  5. Method for nanoscale spatial registration of scanning probes with substrates and surfaces

    NASA Technical Reports Server (NTRS)

    Wade, Lawrence A. (Inventor)

    2010-01-01

    Embodiments in accordance with the present invention relate to methods and apparatuses for aligning a scanning probe used to pattern a substrate, by comparing the position of the probe to a reference location or spot on the substrate. A first light beam is focused on a surface of the substrate as a spatial reference point. A second light beam then illuminates the scanning probe being used for patterning. An optical microscope images both the focused light beam, and a diffraction pattern, shadow, or light backscattered by the illuminated scanning probe tip of a scanning probe microscope (SPM), which is typically the tip of the scanning probe on an atomic force microscope (AFM). Alignment of the scanning probe tip relative to the mark is then determined by visual observation of the microscope image. This alignment process may be repeated to allow for modification or changing of the scanning probe microscope tip.

  6. Comparative in vitro study of tissue welding using a 808 nm diode laser and a Ho:YAG laser.

    PubMed

    Ott, B; Züger, B J; Erni, D; Banic, A; Schaffner, T; Weber, H P; Frenz, M

    2001-01-01

    In vitro porcine arteries and veins have been welded end-to-end using either a 808 nm diode laser combined with an indocyanine green enhanced albumin solder, or with a continuous-wave (cw) Ho:YAG laser without biological solder. The vascular stumps were approached to each other over a coronary dilatation catheter in order to obtain a precise alignment and good coaptation. Standard histology revealed for both welding techniques lateral tissue damage between 2 and 3 mm caused by laser-induced heat. Good solder attachment to the tissue was observed by the use of a scanning electron microscope. The vessels soldered with the 808 nm diode laser using albumin solder showed considerably higher tensile strength (1 N compared to 0.3 N) than vessels welded exclusively by Ho:YAG laser radiation. In contrast, leaking pressure (350 +/- 200 mmHg) and bursting pressure (457 +/- 200 mmHg) were found to be independent of the welding technique used. This study demonstrates that fast (total welding time about 2-5 min), stable and tight microvascular anastomosis can be achieved with the use of a dye-enhanced albumin laser soldering technique and an ancillary coronary dilatation catheter.

  7. Degeneration process of fungiform taste buds after severing the human chorda tympani nerve--observation by confocal laser scanning microscopy.

    PubMed

    Saito, Takehisa; Ito, Tetsufumi; Ito, Yumi; Kato, Yuji; Manabe, Yasuhiro; Narita, Norihiko

    2015-03-01

    To elucidate the degeneration process of fungiform taste buds after severing the chorda tympani nerve (CTN) by confocal laser scanning microscopy in vivo. Prospective study. University hospital. Seven consecutive patients whose CTN was severed during tympanoplasty for middle ear cholesteatoma. Diagnostic. Preoperative and postoperative gustatory functions were assessed by electrogustometry (EGM). An average of 10 fungiform papillae (FP) in the midlateral region of the tongue were periodically observed, and the number of taste buds was counted using a confocal laser microscope. Among them, 2 to 3 reference FPs were selected based on the typical form of the FP or characteristic arrangements of taste pores. Observation was performed before surgery, 1 or 2 days after surgery, 2 or 3 times a week until 2 weeks after surgery, once a week between 2 and 4 weeks, and every 2 to 4 weeks thereafter until all taste buds had disappeared. EGM thresholds showed no response within 1 month after surgery in all patients. The initial change in the degeneration process was the disappearance of taste pores. The surface of taste buds became covered with epithelium. Finally, taste buds themselves atrofied and disappeared. The time course of degeneration differed depending upon individuals, each FP, and each taste bud. By employing the generalized linear mixed model under the Poisson distribution, it was calculated that all taste buds would disappear at around 50 days after surgery. Confocal laser scanning microscopy was useful for clarifying the degeneration process of fungiform taste buds.

  8. Optical tomography of human skin with subcellular spatial and picosecond time resolution using intense near infrared femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Koenig, Karsten; Wollina, Uwe; Riemann, Iris; Peukert, Christiane; Halbhuber, Karl-Juergen; Konrad, Helga; Fischer, Peter; Fuenfstueck, Veronika; Fischer, Tobias W.; Elsner, Peter

    2002-06-01

    We describe the novel high resolution imaging tool DermaInspect 100 for non-invasive diagnosis of dermatological disorders based on multiphoton autofluorescence imaging (MAI)and second harmonic generation. Femtosecond laser pulses in the spectral range of 750 nm to 850 nm have been used to image in vitro and in vivo human skin with subcellular spatial and picosecond temporal resolution. The non-linear induced autofluorescence originates mainly from naturally endogenous fluorophores/protein structures like NAD(P)H, flavins, keratin, collagen, elastin, porphyrins and melanin. Second harmonic generation was observed in the stratum corneum and in the dermis. The system with a wavelength-tunable compact 80 MHz Ti:sapphire laser, a scan module with galvo scan mirrors, piezoelectric objective positioner, fast photon detector and time-resolved single photon counting unit was used to perform optical sectioning and 3D autofluorescence lifetime imaging (t-mapping). In addition, a modified femtosecond laser scanning microscope was involved in autofluorescence measurements. Tissues of patients with psoriasis, nevi, dermatitis, basalioma and melanoma have been investigated. Individual cells and skin structures could be clearly visualized. Intracellular components and connective tissue structures could be further characterized by tuning the excitation wavelength in the range of 750 nm to 850 nm and by calculation of mean fluorescence lifetimes per pixel and of particular regions of interest. The novel non-invasive imaging system provides 4D (x,y,z,t) optical biopsies with subcellular resolution and offers the possibility to introduce a further optical diagnostic method in dermatology.

  9. Fluorescence image-guided photodynamic therapy of cancer cells using a scanning fiber endoscope

    NASA Astrophysics Data System (ADS)

    Woldetensae, Mikias H.; Kirshenbaum, Mark R.; Kramer, Greg M.; Zhang, Liang; Seibel, Eric J.

    2013-03-01

    A scanning fiber endoscope (SFE) and the cancer biomarker 5-aminolevulinic acid (5-ALA) were used to fluorescently detect and destroy superficial cancerous lesions, while experimenting with different dosimetry levels for concurrent or sequential imaging and laser therapy. The 1.6-mm diameter SFE was used to fluorescently image a confluent monolayer of A549 human lung cancer cells from culture, previously administered with 5 mM solution of 5-ALA for 4 hours. Twenty hours after therapy, cell cultures were stained to distinguish between living and dead cells using a laser scanning confocal microscope. To determine relative dosimetry for photodynamic therapy (PDT), 405-nm laser illumination was varied from 1 to 5 minutes with power varying from 5 to 18 mW, chosen to compare equal amounts of energy delivered to the cell culture. The SFE produced 500-line images of fluorescence at 15 Hz using the red detection channel centered at 635 nm. The results show that PDT of A549 cancer cell monolayers using 405nm light for imaging and 5-ALAinduced PpIX therapy was possible using the same SFE system. Increased duration and power of laser illumination produced an increased area of cell death upon live/dead staining. The ultrathin and flexible SFE was able to direct PDT using wide-field fluorescence imaging of a monolayer of cultured cancer cells after uptaking 5-ALA. The correlation between light intensity and duration of PDT was measured. Increased length of exposure and decreased light intensity yields larger areas of cell death than decreased length of exposure with increased light intensity.

  10. Attempt of correlative observation of morphological synaptic connectivity by combining confocal laser-scanning microscope and FIB-SEM for immunohistochemical staining technique.

    PubMed

    Sonomura, Takahiro; Furuta, Takahiro; Nakatani, Ikuko; Yamamoto, Yo; Honma, Satoru; Kaneko, Takeshi

    2014-11-01

    Ten years have passed since a serial block-face scanning electron microscopy (SBF-SEM) method was developed [1]. In this innovative method, samples were automatically sectioned with an ultramicrotome placed inside a scanning electron microscope column, and the block surfaces were imaged one after another by SEM to capture back-scattered electrons. The contrast-inverted images obtained by the SBF-SEM were very similar to those acquired using conventional TEM. SFB-SEM has made easy to acquire image stacks of the transmission electron microscopy (TEM) in the mesoscale, which is taken with the confocal laser-scanning microcopy(CF-LSM).Furthermore, serial-section SEM has been combined with the focused ion beam (FIB) milling method [2]. FIB-incorporated SEM (FIB-SEM) has enabled the acquisition of three-dimensional images with a higher z-axis resolution com- pared to ultramicrotome-equipped SEM.We tried immunocytochemistry for FIB-SEM and correlated this immunoreactivity with that in CF-LSM. Dendrites of neurons in the rat neostriatum were visualized using a recombinant viral vector. Moreover, the thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2). After detection of the sites of terminals apposed to the dendrites by using CF-LSM, GFP and VGluT2 immunoreactivities were further developed for EM by using immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB) methods, respectively.We showed that conventional immuno-cytochemical staining for TEM was applicable to FIB-SEM. Furthermore, several synaptic contacts, which were thought to exist on the basis of CF-LSM findings, were confirmed with FIB-SEM, revealing the usefulness of the combined method of CF-LSM and FIB-SEM. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Pulsed Laser Deposited Ferromagnetic Chromium Dioxide thin Films for Applications in Spintronics

    NASA Astrophysics Data System (ADS)

    Dwivedi, S.; Jadhav, J.; Sharma, H.; Biswas, S.

    Stable rutile type tetragonal chromium dioxide (CrO2) thin films have been deposited on lattice-matched layers of TiO2 by KrF excimer laser based pulsed laser deposition (PLD) technique using Cr2O3 target. The TiO2 seed layer was deposited on oxidized Si substrates by the same PLD process followed by annealing at 1100 °C for 4 h. The lattice-matched interfacial layer is required for the stabilization of Cr (IV) phase in CrO2, since CrO2 behaves as a metastable compound under ambient conditions and readily converts into its stable phase of Cr (III) oxide, Cr2O3. Analyses with X-ray diffraction (XRD), Glancing-angle XRD (GIXRD), Raman spectroscopy and grazing-angle Fourier transform infra-red (FTIR) spectroscopy confirm the presence of tetragonal CrO2 phase in the as-deposited films. Microstructure and surface morphology in the films were studied with field emission scanning electron microscope (FESEM) and atomic force microscope (AFM). Electrical and magnetic characterizations of the films were performed at room temperature. Such type of stable half-metallic CrO2 thin films with low field magnetoresistive switching behaviour are in demand for applications as diverse as spin-FETs, magnetic sensors, and magneto-optical devices.

  12. Yttria-Stabilized Zirconia Ceramic Deposition on SS430 Ferritic Steel Grown by PLD - Pulsed Laser Deposition Method

    NASA Astrophysics Data System (ADS)

    Khalid Rivai, Abu; Mardiyanto; Agusutrisno; Suharyadi, Edi

    2017-01-01

    Development of high temperature materials are one of the key issues for the deployment of advanced nuclear reactors due to higher temperature operation. One of the candidate materials for that purpose is ceramic-coated ferritic steel that one of the functions is to be a thermal barrier coating (TBC). Thin films of YSZ (Ytrria-Stabilized Zirconia) ceramic have been deposited on a SS430 ferritic steel using Pulsed Laser Deposition (PLD) at Center For Science and Technology of Advanced Materials laboratory - National Nuclear Energy Agency of Indonesia (BATAN). The thin film was deposited with the chamber pressure range of 200-225 mTorr, the substrate temperature of 800oC, and the number of laser shots of 3×104, 6×104 and 9×104. Afterward, the samples were analyzed using Scanning Electron Microscope - Energy Dispersive X-ray Spectroscope (SEM-EDS), X-Ray Diffractometer (XRD), Atomic Force Microscope (AFM) and Vickers hardness tester. The results showed that the YSZ could homogeneously and sticky deposited on the surface of the ferritic steel. The surfaces were very smoothly formed with the surface roughness was in the range of 70 nm. Furthermore, thickness, composition of Zr4+ dan Y3+, the crystallinity, and hardness property was increased with the increasing the number of the shots.

  13. Laser Cladding of Ultra-Thin Nickel-Based Superalloy Sheets.

    PubMed

    Gabriel, Tobias; Rommel, Daniel; Scherm, Florian; Gorywoda, Marek; Glatzel, Uwe

    2017-03-10

    Laser cladding is a well-established process to apply coatings on metals. However, on substrates considerably thinner than 1 mm it is only rarely described in the literature. In this work 200 µm thin sheets of nickel-based superalloy 718 are coated with a powder of a cobalt-based alloy, Co-28Cr-9W-1.5Si, by laser cladding. The process window is very narrow, therefore, a precisely controlled Yb fiber laser was used. To minimize the input of energy into the substrate, lines were deposited by setting single overlapping points. In a design of experiments (DoE) study, the process parameters of laser power, laser spot area, step size, exposure time, and solidification time were varied and optimized by examining the clad width, weld penetration, and alloying depth. The microstructure of the samples was investigated by optical microscope (OM) and scanning electron microscopy (SEM), combined with electron backscatter diffraction (EBSD) and energy dispersive X-ray spectroscopy (EDX). Similarly to laser cladding of thicker substrates, the laser power shows the highest influence on the resulting clad. With a higher laser power, the clad width and alloying depth increase, and with a larger laser spot area the weld penetration decreases. If the process parameters are controlled precisely, laser cladding of such thin sheets is manageable.

  14. Laser Cladding of Ultra-Thin Nickel-Based Superalloy Sheets

    PubMed Central

    Gabriel, Tobias; Rommel, Daniel; Scherm, Florian; Gorywoda, Marek; Glatzel, Uwe

    2017-01-01

    Laser cladding is a well-established process to apply coatings on metals. However, on substrates considerably thinner than 1 mm it is only rarely described in the literature. In this work 200 µm thin sheets of nickel-based superalloy 718 are coated with a powder of a cobalt-based alloy, Co–28Cr–9W–1.5Si, by laser cladding. The process window is very narrow, therefore, a precisely controlled Yb fiber laser was used. To minimize the input of energy into the substrate, lines were deposited by setting single overlapping points. In a design of experiments (DoE) study, the process parameters of laser power, laser spot area, step size, exposure time, and solidification time were varied and optimized by examining the clad width, weld penetration, and alloying depth. The microstructure of the samples was investigated by optical microscope (OM) and scanning electron microscopy (SEM), combined with electron backscatter diffraction (EBSD) and energy dispersive X-ray spectroscopy (EDX). Similarly to laser cladding of thicker substrates, the laser power shows the highest influence on the resulting clad. With a higher laser power, the clad width and alloying depth increase, and with a larger laser spot area the weld penetration decreases. If the process parameters are controlled precisely, laser cladding of such thin sheets is manageable. PMID:28772639

  15. The Scanning Optical Microscope: An Overview

    NASA Astrophysics Data System (ADS)

    Kino, G. S.; Corte, T. R.; Xiao, G. Q.

    1988-07-01

    In the last few years there has been a resurgence in research on optical microscopes. One reason stems from the invention of the acoustic microscope by Quate and Lemons,1 and the realization that some of the same principles could be applied to the optical microscope. The acoustic microscope has better transverse definition for the same wavelength than the standard optical microscope and at the same time has far better range definition. Consequently, Kompfner, who was involved with the work on the early acoustic microscope, decided to try out similar scanning microscope principles with optics, and started a group with Wilson and Sheppard to carry out such research at Oxford.2 Sometime earlier, Petran et a13 had invented the tandem scanning microscope which used many of the same principles. Now, in our laboratory at Stanford, these ideas on the tandem scanning microscope and the scanning optical microscope are converging. Another aspect of this work, which stems from the earlier experience with the acoustic microscope, involves measurement of both phase and amplitude of the optical beam. It is also possible to use scanned optical microscopy for other purposes. For instance, an optical beam can be used to excite electrons and holes in semiconductors, and the generated current can be measured. By scanning the optical beam over the semiconductor, an image can be obtained of the regions where there is strong or weak electron hole generation. This type of microscope is called OBIC (Optical Beam Induced Current). A second application involves fluorescent imaging of biological materials. Here we have the excellent range definition of a scanning optical microscope which eliminates unwanted glare from regions of the material where the beam is unfocused.3 A third application is focused on the heating effect of the light beam. With such a system, images can be obtained which are associated with changes in the thermal properties of a material, changes in recombination rates in semiconductors, and differences in material properties associated with either acoustic or thermal effects.4,5 Thus, the range of scanning optical microscopy applications is very large. In the main, the most important applications have been to semiconductors and to biology.

  16. Multimodal optoacoustic and multiphoton fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Sela, Gali; Razansky, Daniel; Shoham, Shy

    2013-03-01

    Multiphoton microscopy is a powerful imaging modality that enables structural and functional imaging with cellular and sub-cellular resolution, deep within biological tissues. Yet, its main contrast mechanism relies on extrinsically administered fluorescent indicators. Here we developed a system for simultaneous multimodal optoacoustic and multiphoton fluorescence 3D imaging, which attains both absorption and fluorescence-based contrast by integrating an ultrasonic transducer into a two-photon laser scanning microscope. The system is readily shown to enable acquisition of multimodal microscopic images of fluorescently labeled targets and cell cultures as well as intrinsic absorption-based images of pigmented biological tissue. During initial experiments, it was further observed that that detected optoacoustically-induced response contains low frequency signal variations, presumably due to cavitation-mediated signal generation by the high repetition rate (80MHz) near IR femtosecond laser. The multimodal system may provide complementary structural and functional information to the fluorescently labeled tissue, by superimposing optoacoustic images of intrinsic tissue chromophores, such as melanin deposits, pigmentation, and hemoglobin or other extrinsic particle or dye-based markers highly absorptive in the NIR spectrum.

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

    NASA Astrophysics Data System (ADS)

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

    2017-05-01

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

  18. Fluorescence correlation spectroscopy, Raster image correlation spectroscopy and Number & Brightness on a commercial confocal laser scanning microscope with analog detectors (Nikon C1)

    PubMed Central

    Moens, Pierre D.J.; Gratton, Enrico; Salvemini, Iyrri L.

    2010-01-01

    Fluorescence correlation spectroscopy (FCS) was developed in 1972 by Magde, Elson and Webb (Magde et al., 1972). Photon counting detectors and avalanche photodiodes have become standards in FCS to the point that there is a widespread belief that these detectors are essential to perform FCS experiments, despite the fact that FCS was developed using analog detectors. Spatial and temporal intensity fluctuation correlations using analog detection on a commercial Olympus Fluoview 300 microscope has been reported by Brown et al. (2008). However, each analog instrument has its own idiosyncrasies that need to be understood before using the instrument for FCS. In this work we explore the capabilities of the Nikon C1, a low cost confocal microscope, to obtain single point FCS, Raster-scan Image Correlation Spectroscopy (RICS) and Number & Brightness data both in solution and incorporated into the membrane of Giant Unilamellar Vesicles (GUVs). We show that it is possible to obtain dynamic information about fluorescent molecules from single point FCS, RICS and Number & Brightness using the Nikon C1. We highlighted the fact that care should be taken in selecting the acquisition parameters in order to avoid possible artifacts due to the detector noise. However, due to relatively large errors in determining the distribution of digital levels for a given microscope setting, the system is probably only adequate for determining relative brightness within the same image. PMID:20734406

  19. Electrical properties of a light-addressable microelectrode chip with high electrode density for extracellular stimulation and recording of excitable cells.

    PubMed

    Bucher, V; Brunner, B; Leibrock, C; Schubert, M; Nisch, W

    2001-05-01

    A light-addressable microelectrode chip with 3600 TiN electrodes was fabricated. Amorphous silicon (a-Si:H) serves as a photo conductor. The electrodes on the chip are addressed by a laser spot and electrical properties of the system are determined. DC measurements show a dark to bright dynamic of 10(6)-10(7). The AC impedance dynamic @ 1 kHz/100 mV and thus the signal-to-noise-ratio is determined to 60. This value is quite sufficient for electrophysiological measurements. For the first time, recordings from cardiac myocytes are reported using the principle of light-addressing. Measurements were done with a standard laser scan microscope (Zeiss LSM 410).

  20. Super-resolution imaging based on the temperature-dependent electron-phonon collision frequency effect of metal thin films

    NASA Astrophysics Data System (ADS)

    Ding, Chenliang; Wei, Jingsong; Xiao, Mufei

    2018-05-01

    We herein propose a far-field super-resolution imaging with metal thin films based on the temperature-dependent electron-phonon collision frequency effect. In the proposed method, neither fluorescence labeling nor any special properties are required for the samples. The 100 nm lands and 200 nm grooves on the Blu-ray disk substrates were clearly resolved and imaged through a laser scanning microscope of wavelength 405 nm. The spot size was approximately 0.80 μm , and the imaging resolution of 1/8 of the laser spot size was experimentally obtained. This work can be applied to the far-field super-resolution imaging of samples with neither fluorescence labeling nor any special properties.

  1. Nanocrystalline ferroelectric BaTiO3/Pt/fused silica for implants synthetized by pulsed laser deposition method

    NASA Astrophysics Data System (ADS)

    Jelínek, Miroslav; Drahokoupil, Jan; Jurek, Karel; Kocourek, Tomáš; Vaněk, Přemysl

    2017-09-01

    The thin-films of BaTiO3 (BTO)/Pt were prepared to test their potential as coatings for titanium-alloy implants. The nanocrystalline BTO/Pt bi-layers were successfully synthesized using fused silica as substrates. The bi-layers were prepared using KrF excimer laser ablation at substrate temperatures (Ts) ranging from 650 °C to 750 °C. The microstructure and composition of the deposits were investigated by scanning electron microscope, x-ray diffraction and wavelength dispersive x-ray spectroscopy methods. The electrical characterization of the Pt/BTO/Pt capacitors indicated ferroelectric-type response in BTO films containing (40-140) nm-sized grains. The technology, microstructure, and functional response of the layers are presented in detail.

  2. UV solid state laser ablation of intraocular lenses

    NASA Astrophysics Data System (ADS)

    Apostolopoulos, A.; Lagiou, D. P.; Evangelatos, Ch.; Spyratou, E.; Bacharis, C.; Makropoulou, M.; Serafetinides, A. A.

    2013-06-01

    Commercially available intraocular lenses (IOLs) are manufactured from silicone and acrylic, both rigid (e.g. PMMA) and foldable (hydrophobic or hydrophilic acrylic biomaterials), behaving different mechanical and optical properties. Recently, the use of apodizing technology to design new diffractive-refractive multifocals improved the refractive outcome of these intraocular lenses, providing good distant and near vision. There is also a major ongoing effort to refine laser refractive surgery to correct other defects besides conventional refractive errors. Using phakic IOLs to treat high myopia potentially provides better predictability and optical quality than corneal-based refractive surgery. The aim of this work was to investigate the effect of laser ablation on IOL surface shaping, by drilling circular arrays of holes, with a homemade motorized rotation stage, and scattered holes on the polymer surface. In material science, the most popular lasers used for polymer machining are the UV lasers, and, therefore, we tried in this work the 3rd and the 5th harmonic of a Q-switched Nd:YAG laser (λ=355 nm and λ=213 nm respectively). The morphology of the ablated IOL surface was examined with a scanning electron microscope (SEM, Fei - Innova Nanoscope) at various laser parameters. Quantitative measurements were performed with a contact profilometer (Dektak-150), in which a mechanical stylus scanned across the surface of gold-coated IOLs (after SEM imaging) to measure variations in surface height and, finally, the ablation rates were also mathematically simulated for depicting the possible laser ablation mechanism(s). The experimental results and the theoretical modelling of UV laser interaction with polymeric IOLs are discussed in relation with the physical (optical, mechanical and thermal) properties of the material, in addition to laser radiation parameters (laser energy fluence, number of pulses). The qualitative aspects of laser ablation at λ=213 nm reveal a smooth optical surface on the intraocular lens with no irregularities, observed with other wavelengths.

  3. Fluorescence lifetime imaging with near-infrared dyes

    NASA Astrophysics Data System (ADS)

    Becker, Wolfgang; Shcheslavskiy, Vladislav

    2013-02-01

    Near-infrared (NIR) dyes are used as fluorescence markers in small-animal imaging and in diffuse optical tomography of the human brain. In these applications it is important to know whether the dyes bind to proteins or other tissue constituents, and whether their fluorescence lifetimes depend on the targets they are bound to. Unfortunately, neither the lasers nor the detectors of commonly used confocal and multiphoton laser scanning microscopes allow for excitation and detection of NIR fluorescence. We therefore upgraded existing confocal TCSPC FLIM systems with NIR lasers and NIR sensitive detectors. In multiphoton systems we used the Ti:Sa laser as a one-photon excitation source in combination with an NIR-sensitive detector in the confocal beam path. We tested a number of NIR dyes in biological tissue. Some of them showed clear lifetime changes depending on the tissue structures they are bound to. We therefore believe that NIR FLIM can deliver supplementary information on the tissue constitution and on local biochemical parameters.

  4. The formation of periodic micro/nano structured on stainless steel by femtosecond laser irradiation

    NASA Astrophysics Data System (ADS)

    Yao, Caizhen; Gao, Wei; Ye, Yayun; Jiang, Yong; Xu, Shizhen; Yuan, Xiaodong

    2017-07-01

    Stainless steel surface was irradiated by linear polarized laser (800 nm, 35 fs, 4 Hz and 0.7 J/cm2) with different pulse numbers. Environmental scanning electron microscope (ESEM/EDS) was used for detailed morphology, microstructure and composition studies. The wettability of irradiated steel surface was tested by Interface Tensiometer JC-2000X and compared with untreated stainless steel. Results showed that micro/nanostripes with different periods were formed. The period increased with the increasing pulse numbers from 450 nm for 90 pulses to 500 nm for 180 pulses. The orientation of those stripes was parallel with the laser beam polarization. Nanoparticles were observed on those periodic structures. EDS indicated that the atomic ratio of Cr increased and the atomic ratios of Fe and Ni decreased after laser irradiation, which may enhance the corrosion resistance due to the Cr-rich layer. The prepared structure exhibited hydrophobic property without further treatment. The formation mechanism of micro/nanoperiodic structures was also explored.

  5. In vivo laser confocal microscopic analysis of corneal K-structures after keratorefractive surgery (LASIK and epi-LASIK).

    PubMed

    Yokogawa, Hideaki; Kobayashi, Akira; Tagawa, Kosaku; Sugiyama, Kazuhisa

    2010-01-01

    To demonstrate alterations of corneal K-structures (sub-Bowman's fibrous structures) after keratorefractive surgery by in vivo laser confocal microscopy and to look for association of K-structures with fluorescein-stained anterior corneal mosaic (ACM). Five patients (nine eyes) participated in this study. For four patients, one eye was evaluated after laser in situ keratomileusis (LASIK) and the other after epipolis-laser in situ keratomileusis (epi-LASIK). For one patient, the left eye was evaluated after epithelial debridement. A photograph of the ACM was obtained. Central corneal regions were scanned by Heidelberg Retina Tomograph 2 Rostock Cornea Module (Heidelberg Engineering GmbH, Heidelberg, Germany). The ACM and K-structures disappeared in all corneas after epi-LASIK, but not after LASIK and epithelial debridement cornea. The presence of K-structures and ACM may be an index to identify eyes that had a previous refractive surgical procedure (surface ablation or LASIK) and be a health index of Bowman layer and adjacent anterior stroma. Copyright 2010, SLACK Incorporated.

  6. Direct Metal Deposition of H13 Tool Steel on Copper Alloy Substrate: Parametric Investigation

    NASA Astrophysics Data System (ADS)

    Imran, M. Khalid; Masood, S. H.; Brandt, Milan

    2015-12-01

    Over the past decade, researchers have demonstrated interest in tribology and prototyping by the laser aided material deposition process. Laser aided direct metal deposition (DMD) enables the formation of a uniform clad by melting the powder to form desired component from metal powder materials. In this research H13 tool steel has been used to clad on a copper alloy substrate using DMD. The effects of laser parameters on the quality of DMD deposited clad have been investigated and acceptable processing parameters have been determined largely through trial-and-error approaches. The relationships between DMD process parameters and the product characteristics such as porosity, micro-cracks and microhardness have been analysed using scanning electron microscope (SEM), image analysis software (ImageJ) and microhardness tester. It has been found that DMD parameters such as laser power, powder mass flow rate, feed rate and focus size have an important role in clad quality and crack formation.

  7. Surface laser alloying of 17-4PH stainless steel steam turbine blades

    NASA Astrophysics Data System (ADS)

    Yao, Jianhua; Wang, Liang; Zhang, Qunli; Kong, Fanzhi; Lou, Chenghua; Chen, Zhijun

    2008-09-01

    As a known high-quality precipitation hardening stainless steel with high strength, high antifatigue, excellent corrosion resistance and good weldability, 17-4PH has been widely used to produce steam turbine blades. However, under the impact of high-speed steam and water droplets, the blades are prone to cavitation, which could lead to lower efficiency, shorter life time, and even accidents. In this article, the 17-4PH blade's surface was alloyed using a high power CO 2 laser. The microstructure and microhardness of hardened 17-4PH were tested by scanning electronic microscope (SEM), X-ray diffraction (XRD), energy disperse spectroscopy (EDS) and a microhardness tester. After laser alloying, the surface layer was denser and the grain refined, while the microhardness of the surface (average 610HV 0.2) was about one times higher than that of the substrate material (330HV 0.2). The friction coefficient of the laser-alloyed 17-4PH layer was much lower than that of the substrate.

  8. Design and performance of a beetle-type double-tip scanning tunneling microscope

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

    Jaschinsky, Philipp; Coenen, Peter; Pirug, Gerhard

    2006-09-15

    A combination of a double-tip scanning tunneling microscope with a scanning electron microscope in ultrahigh vacuum environment is presented. The compact beetle-type design made it possible to integrate two independently driven scanning tunneling microscopes in a small space. Moreover, an additional level for coarse movement allows the decoupling of the translation and approach of the tunneling tip. The position of the two tips can be controlled from the millimeter scale down to 50 nm with the help of an add-on electron microscope. The instrument is capable of atomic resolution imaging with each tip.

  9. Hyperspectral microscopic imaging by multiplex coherent anti-Stokes Raman scattering (CARS)

    NASA Astrophysics Data System (ADS)

    Khmaladze, Alexander; Jasensky, Joshua; Zhang, Chi; Han, Xiaofeng; Ding, Jun; Seeley, Emily; Liu, Xinran; Smith, Gary D.; Chen, Zhan

    2011-10-01

    Coherent anti-Stokes Raman scattering (CARS) microscopy is a powerful technique to image the chemical composition of complex samples in biophysics, biology and materials science. CARS is a four-wave mixing process. The application of a spectrally narrow pump beam and a spectrally wide Stokes beam excites multiple Raman transitions, which are probed by a probe beam. This generates a coherent directional CARS signal with several orders of magnitude higher intensity relative to spontaneous Raman scattering. Recent advances in the development of ultrafast lasers, as well as photonic crystal fibers (PCF), enable multiplex CARS. In this study, we employed two scanning imaging methods. In one, the detection is performed by a photo-multiplier tube (PMT) attached to the spectrometer. The acquisition of a series of images, while tuning the wavelengths between images, allows for subsequent reconstruction of spectra at each image point. The second method detects CARS spectrum in each point by a cooled coupled charged detector (CCD) camera. Coupled with point-by-point scanning, it allows for a hyperspectral microscopic imaging. We applied this CARS imaging system to study biological samples such as oocytes.

  10. Measurement and characterization of external oil in the fried waxy maize starch granules using ATR-FTIR and XRD.

    PubMed

    Chen, Long; Tian, Yaoqi; Sun, Binghua; Cai, Canxin; Ma, Rongrong; Jin, Zhengyu

    2018-03-01

    Concerns regarding increased dietary oil uptake have prompted efforts to investigate the oil absorption and distribution in fried starchy foods. In the present study, attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, together with a chloroform-methanol method, was used to analyze the external and internal oil contents in fried starchy samples. The micromorphology of fried starchy samples was further investigated using scanning electron microscope (SEM), polarized light microscope (PLM) and confocal laser scanning microscopy (CLSM). The results indicated that large amounts of oil were absorbed in or within waxy maize starch, but the majority of oil was located near the surface layer of the starch granules. After defatting, the internal oil was thoroughly removed, while a small amount of external oil remained. As evidenced by the changes of the crystalline characteristics with the help of X-ray diffraction (XRD), the interaction between starch and lipids on the surface was confirmed to form V-type complex compounds during frying at high moisture. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Tip-enhanced near-field Raman spectroscopy with a scanning tunneling microscope and side-illumination optics.

    PubMed

    Yi, K J; He, X N; Zhou, Y S; Xiong, W; Lu, Y F

    2008-07-01

    Conventional Raman spectroscopy (RS) suffers from low spatial resolution and low detection sensitivity due to the optical diffraction limit and small interaction cross sections. It has been reported that a highly localized and significantly enhanced electromagnetic field could be generated in the proximity of a metallic tip illuminated by a laser beam. In this study, a tip-enhanced RS system was developed to both improve the resolution and enhance the detection sensitivity using the tip-enhanced near-field effects. This instrument, by combining RS with a scanning tunneling microscope and side-illumination optics, demonstrated significant enhancement on both optical sensitivity and spatial resolution using either silver (Ag)-coated tungsten (W) tips or gold (Au) tips. The sensitivity improvement was verified by observing the enhancement effects on silicon (Si) substrates. Lateral resolution was verified to be below 100 nm by mapping Ag nanostructures. By deploying the depolarization technique, an apparent enhancement of 175% on Si substrates was achieved. Furthermore, the developed instrument features fast and reliable optical alignment, versatile sample adaptability, and effective suppression of far-field signals.

  12. Surface modification of tooth root canal after application of an X-ray opaque waveguide

    NASA Astrophysics Data System (ADS)

    Dostálová, T.; Jelínková, H.; Šulc, J.; Němec, M.; Koranda, P.; Bartoňová, M.; Radina, P.; Miyagi, M.; Shi, Y.-W.; Matsuura, Y.

    The interest in endodontic use of dental laser systems has been increasing. With the development of thin and flexible delivery systems for various wavelengths, laser applications in endodontics may become even more desirable. The aim of this study is to check the X-ray opacity of a hollow waveguide and to observe the results after laser root canal treatment. The root canal systems of 10 molars were treated endodontically by laser. For the laser radiation source, an Er:YAG laser system generating a wavelength of 2940 nm and an Alexandrite laser system generating a wavelength of 375 nm were used. The hollow waveguide used was checked under X-ray . A root canal surface treated by laser radiation was analyzed by a scanning electron microscope (SEM). The special hollow glass waveguide used was visible in the root canal system under X-ray imaging. Surface modification of the root canal after laser treatment was not found. After conventional treatment the root canal was enlarged. The surface was covered with a smear layer. After application of both laser systems, the smear layer was removed. The resulting canal surface was found to be clean and smooth. Under SEM observation open dentinal tubules were visible. No cracks were present, nor were surface modifications observed.

  13. Benford's Law based detection of latent fingerprint forgeries on the example of artificial sweat printed fingerprints captured by confocal laser scanning microscopes

    NASA Astrophysics Data System (ADS)

    Hildebrandt, Mario; Dittmann, Jana

    2015-03-01

    The possibility of forging latent fingerprints at crime scenes is known for a long time. Ever since it has been stated that an expert is capable of recognizing the presence of multiple identical latent prints as an indicator towards forgeries. With the possibility of printing fingerprint patterns to arbitrary surfaces using affordable ink- jet printers equipped with artificial sweat, it is rather simple to create a multitude of fingerprints with slight variations to avoid raising any suspicion. Such artificially printed fingerprints are often hard to detect during the analysis procedure. Moreover, the visibility of particular detection properties might be decreased depending on the utilized enhancement and acquisition technique. In previous work primarily such detection properties are used in combination with non-destructive high resolution sensory and pattern recognition techniques to detect fingerprint forgeries. In this paper we apply Benford's Law in the spatial domain to differentiate between real latent fingerprints and printed fingerprints. This technique has been successfully applied in media forensics to detect image manipulations. We use the differences between Benford's Law and the distribution of the most significant digit of the intensity and topography data from a confocal laser scanning microscope as features for a pattern recognition based detection of printed fingerprints. Our evaluation based on 3000 printed and 3000 latent print samples shows a very good detection performance of up to 98.85% using WEKA's Bagging classifier in a 10-fold stratified cross-validation.

  14. Evaluation of penetration depth of 2% chlorhexidine digluconate into root dentinal tubules using confocal laser scanning microscope.

    PubMed

    Vadhana, Sekar; Latha, Jothi; Velmurugan, Natanasabapathy

    2015-05-01

    This study evaluated the penetration depth of 2% chlorhexidine digluconate (CHX) into root dentinal tubules and the influence of passive ultrasonic irrigation (PUI) using a confocal laser scanning microscope (CLSM). Twenty freshly extracted anterior teeth were decoronated and instrumented using Mtwo rotary files up to size 40, 4% taper. The samples were randomly divided into two groups (n = 10), that is, conventional syringe irrigation (CSI) and PUI. CHX was mixed with Rhodamine B dye and was used as the final irrigant. The teeth were sectioned at coronal, middle and apical levels and viewed under CLSM to record the penetration depth of CHX. The data were statistically analyzed using Kruskal-Wallis and Mann-Whitney U tests. The mean penetration depths of 2% CHX in coronal, middle and apical thirds were 138 µm, 80 µm and 44 µm in CSI group, respectively, whereas the mean penetration depths were 209 µm, 138 µm and 72 µm respectively in PUI group. Statistically significant difference was present between CSI group and PUI group at all three levels (p < 0.01 for coronal third and p < 0.001 for middle and apical thirds). On intragroup analysis, both groups showed statistically significant difference among three levels (p < 0.001). Penetration depth of 2% CHX into root dentinal tubules is deeper in coronal third when compared to middle and apical third. PUI aided in deeper penetration of 2% CHX into dentinal tubules when compared to conventional syringe irrigation at all three levels.

  15. Efficient and compact Q-switched green laser using graphene oxide as saturable absorber

    NASA Astrophysics Data System (ADS)

    Chang, Jianhua; Li, Hanhan; Yang, Zhenbo; Yan, Na

    2018-01-01

    A new type of graphene oxide (GO) is successfully prepared using an improved modified Hummers method. The Raman shift, X-ray diffraction (XRD), and scanning electron microscope (SEM) measurement techniques are used to characterize the GO. An efficient and compact Q-switched green laser based on Nd:YVO4/PPLN is demonstrated with a few-layered GO as the saturable absorber. Our experimental results show that such a few-layered GO saturable absorber allows for the generation of a stable Q-switched laser pulse centered at 532.1 nm with a 3 dB spectral bandwidth of 2.78 nm, a repetition rate of 71.4 kHz, and a pulse duration of 98 ns. The maximum average output power of 536 mW is obtained at the absorbed pump power of 5.16 W, corresponding to an optical conversion efficiency of 10.3%.

  16. Quasi-crystalline and disordered photonic structures fabricated using direct laser writing

    NASA Astrophysics Data System (ADS)

    Sinelnik, Artem D.; Pinegin, Konstantin V.; Bulashevich, Grigorii A.; Rybin, Mikhail V.; Limonov, Mikhail F.; Samusev, Kirill B.

    2017-09-01

    Direct laser writing is a rapid prototyping technology that has been utilized for the fabrication of micro- and nano-scale materials that have a perfect structure in most of the cases. In this study we exploit the direct laser writing to create several classes of non-periodic materials, such as quasi-crystalline lattices and three-dimensional (3D) objects with an orientation disorder in structural elements. Among quasi-crystalline lattices we consider Penrose tiling and Lévy-type photonic glasses. Images of the fabricated structures are obtained with a scanning electron microscope. In experiment we study the optical diffraction from 3D woodpile photonic structures with orientation disorder and analyze diffraction patters observed on a flat screen positioned behind the sample. With increasing of the disorder degree, we find an impressive transformation of the diffraction patterns from perfect Laue picture to a speckle pattern.

  17. Method for Cleaning Laser-Drilled Holes on Printed Wiring Boards by Plasma Treatment

    NASA Astrophysics Data System (ADS)

    Hirogaki, Toshiki; Aoyama, Eiichi; Minagi, Ryu; Ogawa, Keiji; Katayama, Tsutao; Matsuoka, Takashi; Inoue, Hisahiro

    We propose a new method for cleaning blind via holes after laser drilling of PWBs using oxygen plasma treatment. This report dealt with three kinds of PWB materials: epoxy resin and two kinds of aramid fiber reinforced plastics (AFRP: Technora or Kevlar fiber reinforcement). We observed the drilled holes after plasma treatment using both an optical and a scanning electric microscope (SEM). It was confirmed that adequate etching took place in the drilled holes by plasma treatment. We also compared the hole wall and hole bottom after plasma treatment with ones after chemical etching. It was clear that there was no damage to the aramid fiber tip on the hole wall, and that a smooth roughness of the hole wall was obtained by means of plasma treatment. As a result, we demonstrated that the plasma treatment is effective in cleaning the laser drilled holes of PWBs.

  18. Recycling of pneumatic scrap tyre into nano-crumb rubber by pulsed laser ablation in different pH media

    NASA Astrophysics Data System (ADS)

    Ezaan Khamsan, Nur; Bidin, Noriah; Islam, Shumaila; Daud, Suzairi; Krishnan, Ganesan; Bakar, Mohamad Aizat A.; Naqiuddin Razali, Muhamad; Khamis, Jamil

    2018-05-01

    Nano crumb rubber from scrap tyre is synthesized via 1064 nm pulsed Nd:YAG laser ablation in three different pH media i.e. DI-water (pH∼6.45), D-limonene (pH∼3.47) and NaOH solution (pH∼13.41). Field Emission Scanning Electron Microscope (FESEM) results show spherical morphology of crumb rubber with high degree of aggregation in DI-water and in D-limonene. However, dispersion of crumb rubbers is observed in NaOH solution. The smallest particles size is obtained in NaOH solution within the range of 10.9 nm – 74.3 nm. Energy-dispersive X-ray spectroscopy (EDX) and FTIR analysis confirmed the elements distribution and chemical bonding of rubber with DI-water, D-limonene and NaOH solution. The experimental findings shows that pulsed Nd:YAG laser ablation has potential for fabricating nano-crumb rubber in liquid media.

  19. Preparation, Microstructure and Performance of Nanoscale Ceramics Reinforced Hard Composite Coating

    NASA Astrophysics Data System (ADS)

    Li, Peng

    2014-11-01

    This paper is based on the dry sliding wear of Stellite SF12-B4C-TiN-Mo composite coating deposited on a pure Ti using a laser cladding technique, the parameters of which provide almost crack-free composites with low porosity. To the best of our knowledge, it is the first time that Stellite SF12-B4C-TiN-Mo mixed powders are deposited as the hard composites by a laser cladding technique. Scanning electron microscope images indicate that the nanoscale particles are produced in such coating. The fact that due to the sufficiently rapid heating and cooling rates of the laser cladding technique, the ceramics, such as TiC or TiB2 did not have enough time to grow up, resulting in the formation of the nanoscale particles. Compared with a pure Ti substrate, the increments of the micro-hardness and wear resistance are obtained for such composite coating.

  20. Two temperature approach to femtosecond laser oxidation of molybdenum and morphological study

    NASA Astrophysics Data System (ADS)

    Kotsedi, L.; Kaviyarasu, K.; Fuku, X. G.; Eaton, S. M.; Amara, E. H.; Bireche, F.; Ramponi, R.; Maaza, M.

    2017-11-01

    The two-temperature model was used to gain insight into the thermal evolution of the hot electrons and the crystal lattice of the molybdenum thin coating during femtosecond laser treatment. The heat from the laser raised the bulk temperature of the sample through heat transfer from the hot electron to the crystal lattice of the material, which then led to the melting of the top layer of the film. This process resulted in the hot melt reacting ambient oxygen, which in turn oxidized the surface of molybdenum coating. The topological study and morphology of the oxidized film was conducted using high-resolution scanning electron microscope, with micrographs taken in both the cross-sectional geometry and normal incidence to the electron beam. The molybdenum oxide nanorods were clearly observed and the x-ray diffraction patterns showed the diffraction peaks due to molybdenum oxide.

  1. Nanoscale surface characterization using laser interference microscopy

    NASA Astrophysics Data System (ADS)

    Ignatyev, Pavel S.; Skrynnik, Andrey A.; Melnik, Yury A.

    2018-03-01

    Nanoscale surface characterization is one of the most significant parts of modern materials development and application. The modern microscopes are expensive and complicated tools, and its use for industrial tasks is limited due to laborious sample preparation, measurement procedures, and low operation speed. The laser modulation interference microscopy method (MIM) for real-time quantitative and qualitative analysis of glass, metals, ceramics, and various coatings has a spatial resolution of 0.1 nm for vertical and up to 100 nm for lateral. It is proposed as an alternative to traditional scanning electron microscopy (SEM) and atomic force microscopy (AFM) methods. It is demonstrated that in the cases of roughness metrology for super smooth (Ra >1 nm) surfaces the application of a laser interference microscopy techniques is more optimal than conventional SEM and AFM. The comparison of semiconductor test structure for lateral dimensions measurements obtained with SEM and AFM and white light interferometer also demonstrates the advantages of MIM technique.

  2. Waveguide bends from nanometric silica wires

    NASA Astrophysics Data System (ADS)

    Tong, Limin; Lou, Jingyi; Mazur, Eric

    2005-02-01

    We propose to use bent silica wires with nanometric diameters to guide light as optical waveguide bend. We bend silica wires with scanning tunneling microscope probes under an optical microscope, and wire bends with bending radius smaller than 5 μm are obtained. Light from a He-Ne laser is launched into and guided through the wire bends, measured bending loss of a single bend is on the order of 1 dB. Brief introductions to the optical wave guiding and elastic bending properties of silica wires are also provided. Comparing with waveguide bends based on photonic bandgap structures, the waveguide bends from silica nanometric wires show advantages of simple structure, small overall size, easy fabrication and wide useful spectral range, which make them potentially useful in the miniaturization of photonic devices.

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

    PubMed

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

    2015-10-01

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

  4. Fast scanning mode and its realization in a scanning acoustic microscope

    NASA Astrophysics Data System (ADS)

    Ju, Bing-Feng; Bai, Xiaolong; Chen, Jian

    2012-03-01

    The scanning speed of the two-dimensional stage dominates the efficiency of mechanical scanning measurement systems. This paper focused on a detailed scanning time analysis of conventional raster and spiral scan modes and then proposed two fast alternative scanning modes. Performed on a self-developed scanning acoustic microscope (SAM), the measured images obtained by using the conventional scan mode and fast scan modes are compared. The total scanning time is reduced by 29% of the two proposed fast scan modes. It will offer a better solution for high speed scanning without sacrificing the system stability, and will not introduce additional difficulties to the configuration of scanning measurement systems. They can be easily applied to the mechanical scanning measuring systems with different driving actuators such as piezoelectric, linear motor, dc motor, and so on. The proposed fast raster and square spiral scan modes are realized in SAM, but not specially designed for it. Therefore, they have universal adaptability and can be applied to other scanning measurement systems with two-dimensional mechanical scanning stages, such as atomic force microscope or scanning tunneling microscope.

  5. Swept source optical coherence microscopy using a 1310 nm VCSEL light source

    PubMed Central

    Ahsen, Osman O.; Tao, Yuankai K.; Potsaid, Benjamin M.; Sheikine, Yuri; Jiang, James; Grulkowski, Ireneusz; Tsai, Tsung-Han; Jayaraman, Vijaysekhar; Kraus, Martin F.; Connolly, James L.; Hornegger, Joachim; Cable, Alex; Fujimoto, James G.

    2013-01-01

    We demonstrate high speed, swept source optical coherence microscopy (OCM) using a MEMS tunable vertical cavity surface-emitting laser (VCSEL) light source. The light source had a sweep rate of 280 kHz, providing a bidirectional axial scan rate of 560 kHz. The sweep bandwidth was 117 nm centered at 1310 nm, corresponding to an axial resolution of 13.1 µm in air, corresponding to 8.1 µm (9.6 µm spectrally shaped) in tissue. Dispersion mismatch from different objectives was compensated numerically, enabling magnification and field of view to be easily changed. OCM images were acquired with transverse resolutions between 0.86 µm - 3.42 µm using interchangeable 40X, 20X and 10X objectives with ~600 µm x 600 µm, ~1 mm x 1 mm and ~2 mm x 2 mm field-of-view (FOV), respectively. Parasitic variations in path length with beam scanning were corrected numerically. These features enable swept source OCM to be integrated with a wide range of existing scanning microscopes. Large FOV mosaics were generated by serially acquiring adjacent overlapping microscopic fields and combining them in post-processing. Fresh human colon, thyroid and kidney specimens were imaged ex vivo and compared to matching histology sections, demonstrating the ability of OCM to image tissue specimens. PMID:23938673

  6. Femtosecond laser-induced surface wettability modification of polystyrene surface

    NASA Astrophysics Data System (ADS)

    Wang, Bing; Wang, XinCai; Zheng, HongYu; Lam, YeeCheong

    2016-12-01

    In this paper, we demonstrated a simple method to create either a hydrophilic or hydrophobic surface. With femtosecond laser irradiation at different laser parameters, the water contact angle (WCA) on polystyrene's surface can be modified to either 12.7° or 156.2° from its original WCA of 88.2°. With properly spaced micro-pits created, the surface became hydrophilic probably due to the spread of the water droplets into the micro-pits. While with properly spaced micro-grooves created, the surface became rough and more hydrophobic. We investigated the effect of laser parameters on WCAs and analyzed the laser-treated surface roughness, profiles and chemical bonds by surface profilometer, scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS). For the laser-treated surface with low roughness, the polar (such as C—O, C=O, and O—C=O bonds) and non-polar (such as C—C or C—H bonds) groups were found to be responsible for the wettability changes. While for a rough surface, the surface roughness or the surface topography structure played a more significant role in the changes of the surface WCA. The mechanisms involved in the laser surface wettability modification process were discussed.

  7. Mitochondrial "movement" and lens optics following oxidative stress from UV-B irradiation: cultured bovine lenses and human retinal pigment epithelial cells (ARPE-19) as examples.

    PubMed

    Bantseev, Vladimir; Youn, Hyun-Yi

    2006-12-01

    Mitochondria provide energy generated by oxidative phosphorylation and at the same time play a central role in apoptosis and aging. As a byproduct of respiration, the electron transport chain is known to be the major intracellular site for the generation of reactive oxygen species (ROS). Exposure to solar and occupational ultraviolet (UV) radiation, and thus production of ROS and subsequent cell death, has been implicated in a large spectrum of skin and ocular pathologies, including cataract. Retinal pigment epithelial cell apoptosis generates photoreceptor dysfunction and ultimately visual impairment. The purpose of this article was to characterize in vitro changes following oxidative stress with UV-B radiation in (a) ocular lens optics and cellular function in terms of mitochondrial dynamics of bovine lens epithelium and superficial cortical fiber cells and (b) human retinal pigment epithelial (ARPE-19) cells. Cultured bovine lenses and confluent cultures of ARPE-19 cells were irradiated with broadband UV-B radiation at energy levels of 0.5 and 1.0 J/cm(2). Lens optical function (spherical aberration) was monitored daily up to 14 days using an automated laser scanning system that was developed at the University of Waterloo. This system consists of a single collimated scanning helium-neon laser source that projects a thin (0.05 mm) laser beam onto a plain mirror mounted at 45 degrees on a carriage assembly. This mirror reflects the laser beam directly up through the scanner table surface and through the lens under examination. A digital camera captures the actual position and slope of the laser beam at each step. When all steps have been made, the captured data for each step position is used to calculate the back vertex distance for each position and the difference in that measurement between beams. To investigate mitochondrial movement, the mitochondria-specific fluorescent dye Rhodamine 123 was used. Time series were acquired with a Zeiss 510 (configuration Meta 18) confocal laser scanning microscope equipped with an inverted Axiovert 200 M microscope and 40-x water-immersion C-Apochromat objective (NA 1.2). The optical analysis showed energy level-dependent increases in back vertex distance variability (loss of sharp focus) from 0.39 +/- 0.04 mm (control, n = 11) to 1.63 +/- 0.33 mm (1.0 J/cm(2), n = 10) and 0.63 +/- 0.13 mm (0.5 J/cm(2), n = 9). Confocal laser scanning microscopy analysis of both bovine lenses and ARPE-19 cells showed that following treatment at 0.5 J/cm(2) the mitochondria stopped moving immediately whereas at 1.0 J/cm(2) not only did the mitochondria stop moving, but fragmentation and swelling was seen. Untreated control tissue exhibited up to 15 microm/min of movement of the mitochondria. This could represent normal morphological change, presumably allowing energy transmission across the cell from regions of low to regions of high ATP demand. Lack of mitochondrial movement, fragmentation, and swelling of mitochondria may represent early morphological changes following oxidative stress that may lead to activation of caspase-mediated apoptotic pathways.

  8. Optical track width measurements below 100 nm using artificial neural networks

    NASA Astrophysics Data System (ADS)

    Smith, R. J.; See, C. W.; Somekh, M. G.; Yacoot, A.; Choi, E.

    2005-12-01

    This paper discusses the feasibility of using artificial neural networks (ANNs), together with a high precision scanning optical profiler, to measure very fine track widths that are considerably below the conventional diffraction limit of a conventional optical microscope. The ANN is trained using optical profiles obtained from tracks of known widths, the network is then assessed by applying it to test profiles. The optical profiler is an ultra-stable common path scanning interferometer, which provides extremely precise surface measurements. Preliminary results, obtained with a 0.3 NA objective lens and a laser wavelength of 633 nm, show that the system is capable of measuring a 50 nm track width, with a standard deviation less than 4 nm.

  9. Understanding of the Formation of Micro/Nanoscale Structures on Metal Surfaces by Ultrafast Pulse Laser Processing

    NASA Astrophysics Data System (ADS)

    Peng, Edwin

    In the recent decades, there has been much interest in functionalized surfaces produced by ultrafast laser processing. Using pulse lasers with nanosecond to femtosecond time scale, a wide range of micro/nanoscale structures can be produced on virtually all metal surfaces. These surface structures create special optoelectronic, wetting, and tribological properties with a diverse range of potential applications. The formation mechanisms of these surface structures, especially microscale, mound-like structures, are not fully understood. There has been wide study of ultrafast laser processing of metals. Yet, the proposed formation models present in current literature often lack sufficient experimental verification. Specifically, many studies are limited to surface characterization, e.g. scanning electron microscopy of the surfaces of these micro/nanoscale structures. Valuable insight into the physical processes responsible for formation can be obtained if standard material science characterization methods are performed across the entire mound. In our study, we examined mound-like structures formed on three metal alloys. Using cross section and 3D slice and view operations by a dual beam scanning electron microscope-focused ion beam, the interior microstructures of these mounds are revealed. Taking advantage of amorphous phase formation during laser processing of Ni60Nb40, we verified the fluence-dependent formation model: mounds formed at low fluence are primarily the result of ablation while mounds formed at high fluence are formed by both ablation and rapid resolidification by hydrodynamical fluid flow. For the first time, we revealed the cross section of a wide variety of mound-like structures on titanium surfaces. The increased contribution to mound formation by fluid flow with increasing fluence was observed. Finally, a 3D scanning electron microscopy technique was applied for mounds produced on silver surface by delayed-pulse laser processing. The interior microstructure demonstrated that most of the volume comprised of resolidified silver grains with 1% porosity.

  10. Coherent anti-Stokes Raman scattering spectroscope/microscope based on a widely tunable laser source

    NASA Astrophysics Data System (ADS)

    Dementjev, A.; Gulbinas, V.; Serbenta, A.; Kaucikas, M.; Niaura, G.

    2010-03-01

    We present a coherent anti-Stokes Raman scattering (CARS) microscope based on a robust and simple laser source. A picosecond laser operating in a cavity dumping regime at the 1 MHz repetition rate was used to pump a traveling wave optical parametric generator, which serves as a two-color excitation light source for the CARS microscope. We demonstrate the ability of the presented CARS microscope to measure CARS spectra and images by using several detection schemes.

  11. Morphological and Histological Effects of Radiofrequency and Laser (KTP and Nd:YAG) Treatment of the Inferior Turbinates in Animals.

    PubMed

    Somogyvári, Krisztina; Móricz, Péter; Gerlinger, Imre; Kereskai, László; Szanyi, István; Takács, Ildikó

    2017-02-01

    The aim of this study was to evaluate the short and medium-term effects of radiofrequency (RF) and potassium titanyl phosphate (KTP) and neodymium-yttrium-aluminum garnet (Nd:YAG) laser treatment on the inferior turbinate mucosa in a porcine model. Following randomization, the inferior turbinates were treated either with RF submucosally or with the KTP or the Nd:YAG laser on the surface under videoendoscopic control. Tissue samples were taken at the end of postoperative weeks 1 and 6, and were evaluated macroscopically and histopathologically. Scanning electron microscopy was implemented to demonstrate the morphological changes in the respiratory epithelium. Six weeks following the RF procedure, the mucosa was intact in all cases, and the volume of the inferior turbinates was reduced in the majority of the cases. Although a volume reduction occurred in both laser groups, more complications associated with the healing procedure were noted. With hematoxylin and eosin and periodic acid-Schiff staining, intact epithelium, and submucosal glands remained after the RF procedures at the end of postoperative week 6. Following the KTP-laser intervention, necrotizing sialometaplasia and cartilage destruction occurred, and squamous metaplasia was also apparent in the Nd:YAG group. In both laser groups, dilated glands with excess mucus were seen. The scanning electron microscopic findings demonstrated that cilia were present in all cases. In conclusion, the medium-term macroscopic results were similar in all 3 groups, but the postoperative complications were less following the RF procedure. RF procedure is minimally invasive due to the submucosal intervention that leads to a painless, function preserving recovery.

  12. X-ray laser microscope apparatus

    DOEpatents

    Suckewer, Szymon; DiCicco, Darrell S.; Hirschberg, Joseph G.; Meixler, Lewis D.; Sathre, Robert; Skinner, Charles H.

    1990-01-01

    A microscope consisting of an x-ray contact microscope and an optical microscope. The optical, phase contrast, microscope is used to align a target with respect to a source of soft x-rays. The source of soft x-rays preferably comprises an x-ray laser but could comprise a synchrotron or other pulse source of x-rays. Transparent resist material is used to support the target. The optical microscope is located on the opposite side of the transparent resist material from the target and is employed to align the target with respect to the anticipated soft x-ray laser beam. After alignment with the use of the optical microscope, the target is exposed to the soft x-ray laser beam. The x-ray sensitive transparent resist material whose chemical bonds are altered by the x-ray beam passing through the target mater GOVERNMENT LICENSE RIGHTS This invention was made with government support under Contract No. De-FG02-86ER13609 awarded by the Department of Energy. The Government has certain rights in this invention.

  13. Visible light-harvesting of TiO2 nanotubes array by pulsed laser deposited CdS

    NASA Astrophysics Data System (ADS)

    Bjelajac, Andjelika; Djokic, Veljko; Petrovic, Rada; Socol, Gabiel; Mihailescu, Ion N.; Florea, Ileana; Ersen, Ovidiu; Janackovic, Djordje

    2014-08-01

    Titanium dioxide (TiO2) nanotubes arrays, obtained by anodization technique and annealing, were decorated with CdS using pulsed laser deposition method. Their structural, morphological and chemical characterization was carried out by electron microscopy in scanning (SEM) and transmission (TEM) modes, combined with energy dispersive spectroscopy (EDS) and electron energy loss spectroscopy (EELS). It was demonstrated that the quantity of deposited CdS can be controlled by varying the number of laser pulses. The chemical mapping of the elements of interest was performed using the energy filtered mode of the electron microscope. The results showed that pulse laser deposition is an adequate technique for deposition of CdS inside and between 100 nm wide TiO2 nanotubes. The diffuse reflectance spectroscopy investigation of selected samples proved that the absorption edge of the prepared CdS/TiO2 nanocomposites is significantly extended to the visible range. The corresponding band gaps were determinated from the Tauc plot of transformed Kubelka-Munk function. The band gap reduction of TiO2 nanotubes by pulsed laser deposition of CdS was put in evidence.

  14. Laser-Based Surface Modification of Microstructure for Carbon Fiber-Reinforced Plastics

    NASA Astrophysics Data System (ADS)

    Yang, Wenfeng; Sun, Ting; Cao, Yu; Li, Shaolong; Liu, Chang; Tang, Qingru

    2018-05-01

    Bonding repair is a powerful feature of carbon fiber-reinforced plastics (CFRP). Based on the theory of interface bonding, the interface adhesion strength and reliability of the CFRP structure will be directly affected by the microscopic features of the CFRP surface, including the microstructure, physical, and chemical characteristics. In this paper, laser-based surface modification was compared to Peel-ply, grinding, and polishing to comparatively evaluate the surface microstructure of CFRP. The surface microstructure, morphology, fiber damage, height and space parameters were investigated by scanning electron microscopy (SEM) and laser confocal microscopy (LCM). Relative to the conventional grinding process, laser modification of the CFRP surface can result in more uniform resin removal and better processing control and repeatability. This decreases the adverse impact of surface fiber fractures and secondary damage. The surface properties were significantly optimized, which has been reflected such things as the obvious improvement of surface roughness, microstructure uniformity, and actual area. The improved surface microstructure based on laser modification is more conducive to interface bonding of CFRP structure repair. This can enhance the interfacial adhesion strength and reliability of repair.

  15. Experimental investigations of the use of an erbium:YAG laser on temporomandibular joint (TMJ) structures: first experimental results

    NASA Astrophysics Data System (ADS)

    Nuebler-Moritz, Michael; Niederdellmann, Herbert; Hering, Peter; Deuerling, Christian; Dammer, Ralf; Behr, M.

    1995-04-01

    The following paper introduces the results of an interdisciplinary research project. With the aid of photomacroscopic examination, light and scanning electron microscope investigations, changes to temporomandibular joint structures were detected in vitro after irradiation with an Erbium:YAG laser system. The solid-state Erbium:YAG laser, operating at a wavelength of 2.94 micrometers was used in the normal- spiking mode. The free-running laser beam was focussed onto freshly excised porcine tissue samples using a 108-mm sapphire lens. In this study the output was generally pulsed at a repetition rate of 4 Hz, with a pulse duration varying from 120 microsecond(s) to 500 microsecond(s) . Between 50 mJ and 500 mJ per pulse were applied to create pinpoint lesions. The optimum average energy density and pulse duration of the Erbium:YAG laser radiation for the purpose of TMJ-surgery (as far as it concerns meniscus and articulating facets) - which means efficient etch rate and minimal adjacent injury - seems to be about 24-42 J/cm2 and 120 microsecond(s) -240 microsecond(s) , respectively.

  16. Laser cutting of various materials: Kerf width size analysis and life cycle assessment of cutting process

    NASA Astrophysics Data System (ADS)

    Yilbas, Bekir Sami; Shaukat, Mian Mobeen; Ashraf, Farhan

    2017-08-01

    Laser cutting of various materials including Ti-6Al-4V alloy, steel 304, Inconel 625, and alumina is carried out to assess the kerf width size variation along the cut section. The life cycle assessment is carried out to determine the environmental impact of the laser cutting in terms of the material waste during the cutting process. The kerf width size is formulated and predicted using the lump parameter analysis and it is measured from the experiments. The influence of laser output power and laser cutting speed on the kerf width size variation is analyzed using the analytical tools including scanning electron and optical microscopes. In the experiments, high pressure nitrogen assisting gas is used to prevent oxidation reactions in the cutting section. It is found that the kerf width size predicted from the lump parameter analysis agrees well with the experimental data. The kerf width size variation increases with increasing laser output power. However, this behavior reverses with increasing laser cutting speed. The life cycle assessment reveals that material selection for laser cutting is critical for the environmental protection point of view. Inconel 625 contributes the most to the environmental damages; however, recycling of the waste of the laser cutting reduces this contribution.

  17. Evidence of a rolling motion of a microparticle on a silicon wafer in a liquid environment

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

    Schiwek, Simon; Stark, Robert W., E-mail: stark@csi.tu-darmstadt.de, E-mail: dietz@csi.tu-darmstadt.de; Dietz, Christian, E-mail: stark@csi.tu-darmstadt.de, E-mail: dietz@csi.tu-darmstadt.de

    2016-05-21

    The interaction of micro- and nanometer-sized particles with surfaces plays a crucial role when small-scale structures are built in a bottom-up approach or structured surfaces are cleaned in the semiconductor industry. For a reliable quantification of the interaction between individual particles and a specific surface, however, the motion type of the particle must be known. We developed an approach to unambiguously distinguish between sliding and rolling particles. To this end, fluorescent particles were partially bleached in a confocal laser scanning microscope to tailor an optical inhomogeneity, which allowed for the identification of the characteristic motion pattern. For the manipulation, themore » water flow generated by a fast moving cantilever-tip of an atomic force microscope enabled the contactless pushing of the particle. We thus experimentally evidenced a rolling motion of a micrometer-sized particle directly with a fluorescence microscope. A similar approach could help to discriminate between rolling and sliding particles in liquid flows of microfluidic systems.« less

  18. Scanning evanescent electro-magnetic microscope

    DOEpatents

    Xiang, Xiao-Dong; Gao, Chen; Schultz, Peter G.; Wei, Tao

    2003-01-01

    A novel scanning microscope is described that uses near-field evanescent electromagnetic waves to probe sample properties. The novel microscope is capable of high resolution imaging and quantitative measurements of the electrical properties of the sample. The inventive scanning evanescent wave electromagnetic microscope (SEMM) can map dielectric constant, tangent loss, conductivity, complex electrical impedance, and other electrical parameters of materials. The quantitative map corresponds to the imaged detail. The novel microscope can be used to measure electrical properties of both dielectric and electrically conducting materials.

  19. Scanning evanescent electro-magnetic microscope

    DOEpatents

    Xiang, Xiao-Dong; Gao, Chen

    2001-01-01

    A novel scanning microscope is described that uses near-field evanescent electromagnetic waves to probe sample properties. The novel microscope is capable of high resolution imaging and quantitative measurements of the electrical properties of the sample. The inventive scanning evanescent wave electromagnetic microscope (SEMM) can map dielectric constant, tangent loss, conductivity, complex electrical impedance, and other electrical parameters of materials. The quantitative map corresponds to the imaged detail. The novel microscope can be used to measure electrical properties of both dielectric and electrically conducting materials.

  20. PScan 1.0: flexible software framework for polygon based multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Li, Yongxiao; Lee, Woei Ming

    2016-12-01

    Multiphoton laser scanning microscopes exhibit highly localized nonlinear optical excitation and are powerful instruments for in-vivo deep tissue imaging. Customized multiphoton microscopy has a significantly superior performance for in-vivo imaging because of precise control over the scanning and detection system. To date, there have been several flexible software platforms catered to custom built microscopy systems i.e. ScanImage, HelioScan, MicroManager, that perform at imaging speeds of 30-100fps. In this paper, we describe a flexible software framework for high speed imaging systems capable of operating from 5 fps to 1600 fps. The software is based on the MATLAB image processing toolbox. It has the capability to communicate directly with a high performing imaging card (Matrox Solios eA/XA), thus retaining high speed acquisition. The program is also designed to communicate with LabVIEW and Fiji for instrument control and image processing. Pscan 1.0 can handle high imaging rates and contains sufficient flexibility for users to adapt to their high speed imaging systems.

  1. Relationship between gustatory function and average number of taste buds per fungiform papilla measured by confocal laser scanning microscopy in humans.

    PubMed

    Saito, Takehisa; Ito, Tetsufumi; Ito, Yumi; Manabe, Yasuhiro; Sano, Kazuo

    2017-02-01

    The aim of this study was to elucidate the relationship between the gustatory function and average number of taste buds per fungiform papilla (FP) in humans. Systemically healthy volunteers (n = 211), pre-operative patients with chronic otitis media (n = 79), and postoperative patients, with or without a chorda tympani nerve (CTN) severed during middle ear surgery (n = 63), were included. Confocal laser scanning microscopy was employed to observe fungiform taste buds because it allows many FP to be observed non-invasively in a short period of time. Taste buds in an average of 10 FP in the midlateral region of the tongue were counted. In total, 3,849 FP were observed in 353 subjects. The gustatory function was measured by electrogustometry (EGM). An inverse relationship was found between the gustatory function and average number of fungiform taste buds per papilla. The healthy volunteers showed a lower EGM threshold (better gustatory function) and had more taste buds than did the patients with otitis media, and the patients with otitis media showed a lower EGM threshold and had more taste buds than did postoperative patients, reflecting the severity of damage to the CTN. It was concluded that the confocal laser scanning microscope is a very useful tool for using to observe a large number of taste buds non-invasively. © 2017 Eur J Oral Sci.

  2. Dye-Assisted Laser Skin Closure with Pulsed Radiation: An In Vitro Study of Weld Strength and Thermal Damage

    NASA Astrophysics Data System (ADS)

    Fried, Nathaniel M.; Walsh, Joseph T.

    1998-10-01

    Previous laser skin welding studies have used continuous wave delivery of radiation. However, heat diffusion during irradiation prevents strong welds from being achieved without creating large zones of thermal damage. Previously published results indicate that a thermal damage zone in skin greater than 200 micrometers may prevent normal wound healing. We proposed that both strong welds and minimal thermal damage can be achieved by introducing a dye and delivering the radiation in a series of sufficiently short pulses. Two-cm-long incisions were made in guinea pig skin, in vitro. India ink and egg white (albumin) were applied to the wound edges to enhance radiation absorption and to close the wound, respectively. Continuous wave (cw), 1.06 micrometers , Nd:yttrium-aluminum-garnet laser radiation was scanned over the weld producing approximately 100 ms pulses. The cooling time between scans and the number of scans was varied. The thermal damage zone at the weld edges was measured using a transmission polarizing light microscope. The tensile strength of the welds was measured using a tensiometer. For pulsed welding and long cooling times between pulses (8 s), weld strengths of 2.4 +/- 0.9 kg/cm2 were measured, and lateral thermal damage at the epidermis was limited to 500 +/- 150 micrometers . With cw welding, comparable weld strengths produced 2700 +/- 300 micrometers of lateral thermal damage. The cw weld strengths were only 0.6 +/- 0.3 kg/cm2 for thermal damage zones comparable to pulsed welding.

  3. Rapid fibrin plug formation within cutaneous ablative fractional CO2 laser lesions.

    PubMed

    Kositratna, Garuna; Evers, Michael; Sajjadi, Amir; Manstein, Dieter

    2016-02-01

    Ablative fractional laser procedures have been shown to facilitate topical drug delivery into the skin. Past studies have mainly used ex vivo models to demonstrate enhanced drug delivery and in vivo studies have investigated laser created channels over a time course of days and weeks rather than within the first few minutes and hours after exposures. We have noticed rapid in vivo fibrin plug formation within ablative fractional laser lesions impairing passage through the laser created channels. In vivo laser exposures were performed in a porcine model. A fractional CO2 laser (AcuPulse™ system, AcuScan 120™ handpiece, Lumenis, Inc., Yokneam, Israel) was programmed in quasi-continuous wave (QCW) mode, at 40W, 50 mJ per pulse, 5% coverage, nominal 120 µm spot size, 8 × 8 mm square pattern, 169 MTZs per scan. Six millimeters punch biopsies were procured at 0, 2, 5, 10, 15, 30, 60, 90 minutes after completion of each scan, then fixed in 10% formalin. 12 repeats were performed of each time point. Skin samples were processed for serial vertically cut paraffin sections (5 μm collected every 25 μm) then H&E and special immunohistochemistry staining for fibrin and platelet. Dimensions of Microscopic Treatment Zones (MTZs) and extent of fibrin plug were assessed and quantified histologically. Ex vivo laser exposures of the identical laser parameter were performed on porcine and human skin at different storage conditions. Histology procured at various predetermined time intervals after in vivo fractional CO2 laser exposures revealed a rapidly forming fibrin plug initiating at the bottom of the MTZ lesions. At longer time intervals, the fibrin plug was extending towards the superficial sections. Within the first 5 minutes, more than 25% length of the entire laser-ablated channel was filled with a fibrin plug. With increased time intervals, the cavity was progressively filled with a fibrin plug. At 90 minutes, more than 90% length of the entire laser-ablated channel was occluded. Ex vivo exposures failed to produce any significant fibrin plug formation. The current study has demonstrated rapid fibrin plug formation after ablative fractional laser procedures. It was shown that the passage through laser created pathways is critically time dependent for in vivo exposures. In contrast, ex vivo exposures do not exhibit such time dependent passage capacity. In particular, drug, substance, and cell delivery studies for ablative fractional laser treatments should take early fibrin plug formation into consideration and further investigate the impact on transdermal delivery. © 2015 Wiley Periodicals, Inc.

  4. Effects of erbium-and chromium-doped yttrium scandium gallium garnet and diode lasers on the surfaces of restorative dental materials: a scanning electron microscope study.

    PubMed

    Hatipoglu, M; Barutcigil, C

    2015-01-01

    The aim of this study is to evaluate the potential effects of laser irradiation, which is commonly performed in periodontal surgery, on the surfaces of restorative materials. Five different restorative dental materials were used in this study, as follows: (1) Resin composite, (2) poly acid-modified resin composite (compomer), (3) conventional glass ionomer cement (GIC), (4) resin-modified glass ionomer cement (RMGIC), and (5) amalgam. Four cylindrical samples (8 mm diameter, 2 mm height) were prepared for each restorative material. In addition, four freshly extracted, sound human incisors teeth were selected. Two different laser systems commonly used in periodontal surgery were examined in this study: A 810 nm diode laser at a setting of 1 W with continuous-phase laser irradiation for 10 s, and an erbium-and chromium-doped yttrium scandium gallium garnet (Er, Cr: YSGG) laser at settings of 2.5 W, 3.25 W, and 4 W with 25 Hz laser irradiation for 10 s. Scanning electron microscopy (SEM) analysis was performed to evaluate the morphology and surface deformation of the restorative materials and tooth surfaces. According to the SEM images, the Er, Cr: YSGG laser causes irradiation markings that appear as demineralized surfaces on tooth samples. The Er, Cr: YSGG laser also caused deep defects on composite, compomer, and RMGIC surfaces because of its high power, and the ablation was deeper for these samples. High-magnification SEM images of GIC samples showed the melting and combustion effects of the Er, Cr: YSGG laser, which increased as the laser power was increased. In amalgam samples, neither laser left significant harmful effects at the lowest power setting. The diode laser did cause irradiation markings, but they were insignificant compared with those left by the Er, Cr: YSGG laser on the surfaces of the different materials and teeth. Within the limitations of this study, it can be concluded that Er, Cr: YSGG laser irradiation could cause distortions of the surfaces of restorative materials. Diode lasers can be preferred for periodontal surgery.

  5. Silicon solar cell fabrication technology

    NASA Technical Reports Server (NTRS)

    Stafsudd, O. M.

    1979-01-01

    The laser cell scanner was used to characterize a number of solar cells made in various materials. An electron beam-induced current (EBIC) study was performed using a stereoscan scanning electron microscope. Planar p-n junctions were analyzed. A theory for the EBIC based on the analytical solution of the ambipolar diffusion equation under the influence of electron beam excitation parameter z (which is related to beam penetration), the junction depth Z sub j, the beam current and the surface recombination, was formulated and tested. The effect of a grain boundary was studied.

  6. Nonlinear optical and microscopic analysis of Cu2+ doped zinc thiourea chloride (ZTC) monocrystal

    NASA Astrophysics Data System (ADS)

    Ramteke, S. P.; Anis, Mohd; Pandian, M. S.; Kalainathan, S.; Baig, M. I.; Ramasamy, P.; Muley, G. G.

    2018-02-01

    Organometallic crystals offer considerable nonlinear response therefore, present article focuses on bulk growth and investigation of Cu2+ ion doped zinc thiourea chloride (ZTC) crystal to explore its technological impetus for laser assisted nonlinear optical (NLO) device applications. The Cu2+ ion doped ZTC bulk single crystal of dimension 03 × 2.4 × 0.4 cm3 has been grown from pH controlled aqueous solution by employing slow solvent evaporation technique. The structural analysis has been performed by means of single crystal X-ray diffraction technique. The doping of Cu2+ ion in ZTC crystal matrix has been confirmed by means of energy dispersive spectroscopic (EDS) technique. The origin of nonlinear optical properties in Cu2+ ion doped ZTC crystal has been studied by employing the Kurtz-Perry test and Z-scan analysis. The remarkable enhancement in second harmonic generation (SHG) efficiency of Cu2+ ion doped ZTC crystal with reference to ZTC crystal has been determined. The He-Ne laser assisted Z-scan analysis has been performed to determine the third order nonlinear optical (TONLO) nature of grown crystal. The TONLO parameters such as susceptibility, absorption coefficient, refractive index and figure of merit of Cu-ZTC crystal have been evaluated using the Z-scan transmittance data. The laser damage threshold of grown crystal to high intensity of Nd:YAG laser is found to be 706.2 MW/cm2. The hardness number, work hardening index, yield strength and elastic stiffness coefficient of grown crystal has been investigated under microhardness study. The etching study has been carried out to determine the growth likelihood, nature of etch pits and surface quality of grown crystal.

  7. Theory of a Quantum Scanning Microscope for Cold Atoms

    NASA Astrophysics Data System (ADS)

    Yang, D.; Laflamme, C.; Vasilyev, D. V.; Baranov, M. A.; Zoller, P.

    2018-03-01

    We propose and analyze a scanning microscope to monitor "live" the quantum dynamics of cold atoms in a cavity QED setup. The microscope measures the atomic density with subwavelength resolution via dispersive couplings to a cavity and homodyne detection within the framework of continuous measurement theory. We analyze two modes of operation. First, for a fixed focal point the microscope records the wave packet dynamics of atoms with time resolution set by the cavity lifetime. Second, a spatial scan of the microscope acts to map out the spatial density of stationary quantum states. Remarkably, in the latter case, for a good cavity limit, the microscope becomes an effective quantum nondemolition device, such that the spatial distribution of motional eigenstates can be measured backaction free in single scans, as an emergent quantum nondemolition measurement.

  8. Theory of a Quantum Scanning Microscope for Cold Atoms.

    PubMed

    Yang, D; Laflamme, C; Vasilyev, D V; Baranov, M A; Zoller, P

    2018-03-30

    We propose and analyze a scanning microscope to monitor "live" the quantum dynamics of cold atoms in a cavity QED setup. The microscope measures the atomic density with subwavelength resolution via dispersive couplings to a cavity and homodyne detection within the framework of continuous measurement theory. We analyze two modes of operation. First, for a fixed focal point the microscope records the wave packet dynamics of atoms with time resolution set by the cavity lifetime. Second, a spatial scan of the microscope acts to map out the spatial density of stationary quantum states. Remarkably, in the latter case, for a good cavity limit, the microscope becomes an effective quantum nondemolition device, such that the spatial distribution of motional eigenstates can be measured backaction free in single scans, as an emergent quantum nondemolition measurement.

  9. Imaging ion and molecular transport at subcellular resolution by secondary ion mass spectrometry

    NASA Astrophysics Data System (ADS)

    Chandra, Subhash; Morrison, George H.

    1995-05-01

    The transport of K+, Na+, and Ca2+ were imaged in individual cells with a Cameca IMS-3f ion microscope. Strict cryogenic frozen freeze-dry sample preparations were employed. Ion redistribution artifacts in conventional chemical preparations are discussed. Cryogenically prepared freeze-fractured freeze-dried cultured cells allowed the three-dimensional ion microscopic imaging of elements. As smaller structures in calcium images can be resolved with the 0.5 [mu]m spatial resolution, correlative techniques are needed to confirm their identity. The potentials of reflected light microscopy, scanning electron microscopy and laser scanning confocal microscopy are discussed for microfeature recognition in freeze-fractured freeze-dried cells. The feasibility of using frozen freeze-dried cells for imaging molecular transport at subcellular resolution was tested. Ion microscopy successfully imaged the transport of the isotopically tagged (13C, 15N) amino acid, -arginine. The labeled amino acid was imaged at mass 28 with a Cs+ primary ion beam as the 28(13C15N)- species. After a 4 h exposure of LLC-PK1 kidney cells to 4 mM labeled arginine, the amino acid was localized throughout the cell with a preferential incorporation into the nucleus and nucleolus. An example is also shown of the ion microscopic imaging of sodium borocaptate, an experimental therapeutic drug for brain tumors, in cryogenically prepared frozen freeze-dried Swiss 3T3 cells.

  10. Epithelial innervation of human cornea: a three-dimensional study using confocal laser scanning fluorescence microscopy.

    PubMed

    Guthoff, Rudolf F; Wienss, Holger; Hahnel, Christian; Wree, Andreas

    2005-07-01

    Evaluation of a new method to visualize distribution and morphology of human corneal nerves (Adelta- and C-fibers) by means of fluorescence staining, confocal laser scanning microscopy, and 3-dimensional (3D) reconstruction. Trephinates of corneas with a diagnosis of Fuchs corneal dystrophy were sliced into layers of 200 microm thickness using a Draeger microkeratome (Storz, Germany). The anterior lamella was stained with the Life/Dead-Kit (Molecular Probes Inc.), examined by the confocal laser scanning microscope "Odyssey XL," step size between 0.5 and 1 microm, and optical sections were digitally 3D-reconstructed. Immediate staining of explanted corneas by the Life/Dead-Kit gave a complete picture of the nerves in the central human cornea. Thin nerves running parallel to the Bowman layer in the subepithelial plexus perforate the Bowman layer orthogonally through tube-like structures. Passing the Bowman layer, Adelta- and C-fibers can be clearly distinguished by fiber diameter, and, while running in the basal epithelial plexus, by their spatial arrangement. Adelta-fibers run straight and parallel to the Bowman layer underneath the basal cell layer. C-fibers, after a short run parallel to the Bowman layer, send off multiple branches penetrating epithelial cell layers orthogonally, ending blindly in invaginations of the superficial cells. In contrast to C-fibers, Adelta-fibers show characteristic bulbous formations when kinking into the basal epithelial plexus. Ex-vivo fluorescence staining of the cornea and 3D reconstructions of confocal scans provide a fast and easily reproducible tool to visualize nerves of the anterior living cornea at high resolution. This may help to clarify gross variations of nerve fiber patterns under various clinical and experimental conditions.

  11. Laser irradiation effects on the surface, structural and mechanical properties of Al-Cu alloy 2024

    NASA Astrophysics Data System (ADS)

    Yousaf, Daniel; Bashir, Shazia; Akram, Mahreen; kalsoom, Umm-i.-; Ali, Nisar

    2014-02-01

    Laser irradiation effects on surface, structural and mechanical properties of Al-Cu-Mg alloy (Al-Cu alloy 2024) have been investigated. The specimens were irradiated for various fluences ranging from 3.8 to 5.5 J/cm2 using an Excimer (KrF) laser (248 nm, 18 ns, 30 Hz) under vacuum environment. The surface and structural modifications of the irradiated targets have been investigated by scanning electron microscope (SEM) and X-ray diffractometer (XRD), respectively. SEM analysis reveals the formation of micro-sized craters along the growth of periodic surface structures (ripples) at their peripheries. The size of the craters initially increases and then decreases by increasing the laser fluence. XRD analysis shows an anomalous trend in the peak intensity and crystallite size of the specimen irradiated for various fluences. A universal tensile testing machine and Vickers microhardness tester were employed in order to investigate the mechanical properties of the irradiated targets. The changes in yield strength, ultimate tensile strength and microhardness were found to be anomalous with increasing laser fluences. The changes in the surface and structural properties of Al-Cu alloy 2024 after laser irradiation have been associated with the changes in mechanical properties.

  12. Analysis of the Microstructure and Thermal Shock Resistance of Laser Glazed Nanostructured Zirconia TBCs

    NASA Astrophysics Data System (ADS)

    Chen, Hui; Hao, Yunfei; Wang, Hongying; Tang, Weijie

    2010-03-01

    Nanostructured zirconia thermal barrier coatings (TBCs) have been prepared by atmospheric plasma spraying using the reconstituted nanosized yttria partially stabilized zirconia powder. Field emission scanning electron microscope was applied to examine the microstructure of the resulting TBCs. The results showed that the TBCs exhibited a unique, complex structure including nonmelted or partially melted nanosized particles and columnar grains. A CO2 continuous wave laser beam has been applied to laser glaze the nanostructured zirconia TBCs. The effect of laser energy density on the microstructure and thermal shock resistance of the as-glazed coatings has been systematically investigated. SEM observation indicated that the microstructure of the as-glazed coatings was very different from the microstructure of the as-sprayed nanostructured TBCs. It changed from single columnar grain to a combination of columnar grains in the fracture surface and equiaxed grains on the surface with increasing laser energy density. Thermal shock resistance tests have showed that laser glazing can double the lifetime of TBCs. The failure of the as-glazed coatings was mainly due to the thermal stress caused by the thermal expansion coefficient mismatch between the ceramic coat and metallic substrate.

  13. Surface modification of AISI H13 tool steel by laser cladding with NiTi powder

    NASA Astrophysics Data System (ADS)

    Norhafzan, B.; Aqida, S. N.; Chikarakara, E.; Brabazon, D.

    2016-04-01

    This paper presents laser cladding of NiTi powder on AISI H13 tool steel surface for surface properties enhancement. The cladding process was conducted using Rofin DC-015 diffusion-cooled CO2 laser system with wavelength of 10.6 µm. NiTi powder was pre-placed on H13 tool steel surface. The laser beam was focused with a spot size of 90 µm on the sample surface. Laser parameters were set to 1515 and 1138 W peak power, 18 and 24 % duty cycle and 2300-3500 Hz laser pulse repetition frequency. Hardness properties of the modified layer were characterized by Wilson Hardness tester. Metallographic study and chemical composition were conducted using field emission scanning electron microscope and energy-dispersive X-ray spectrometer (EDXS) analysis. Results showed that hardness of NiTi clad layer increased three times that of the substrate material. The EDXS analysis detected NiTi phase presence in the modified layer up to 9.8 wt%. The metallographic study shows high metallurgical bonding between substrate and modified layer. These findings are significant to both increased hardness and erosion resistance of high-wear-resistant components and elongating their lifetime.

  14. Partial ablation of stratum corneum by UV (193-nm) or IR (2.94-μm) pulsed lasers to enhance transdermal drug delivery rate

    NASA Astrophysics Data System (ADS)

    Fujiwara, Ai; Hinokitani, Toshihiro; Goto, Kenichi; Arai, Tsunenori

    2004-07-01

    To develop the noninvasive transdermal drug delivery system, pulsed lasers (argon-fluoride excimer laser (ArF laser) and erbium:yittrium aluminum garnet laser (Er:YAG laser)) were used to partially ablate the stratum corneum (SC), the upper layer of the skin. Because of the barrier function of the SC to drug permeation, the number of drugs especially macromolecules used in transdermal drug delivery system without skin irritation has been limited. Ultrastructural changes on the SC surface of ablated Yucatan micropig skin in vitro were observed with Environmental Scanning Electron Microscope. The result indicated that the structural changes varied according to each laser sources and irradiation conditions (laser fluences and numbers of pulses). Many granular structures of about 2 μm in diameter were observed in the ablated sites on ArF laser with lower fluence exposure (30 mJ/cm2, 200 pulses), and plane structures in the sites with higher fluence exposure (80 mJ/cm2, 80 pulses). In contrast, the ablation of Er:YAG laser created some pores of about 20 μm across on the surface of the SC. Under the irradiation condition of partial ablation, the skin permeability of macromolecule compound was enhanced. This partial SC ablation by pulsed laser could be possible candidate of the noninvasive transdermal drug delivery system with good physiological conditions of skin.

  15. Fs-laser ablation of teeth is temperature limited and provides information about the ablated components.

    PubMed

    de Menezes, Rebeca Ferraz; Harvey, Catherine Malinda; de Martínez Gerbi, Marleny Elizabeth Márquez; Smith, Zachary J; Smith, Dan; Ivaldi, Juan C; Phillips, Alton; Chan, James W; Wachsmann-Hogiu, Sebastian

    2017-10-01

    The goal of this work is to investigate the thermal effects of femtosecond laser (fs-laser) ablation for the removal of carious dental tissue. Additional studies identify different tooth tissues through femtosecond laser induced breakdown spectroscopy (fsLIBS) for the development of a feedback loop that could be utilized during ablation in a clinical setting. Scanning Election Microscope (SEM) images reveal that minimal morphological damages are incurred at repetition rates below the carbonization threshold of each tooth tissue. Thermal studies measure the temperature distribution and temperature decay during laser ablation and after laser cessation, and demonstrate that repetition rates at or below 10kHz with a laser fluence of 40 J/cm 2 would inflict minimal thermal damage on the surrounding nerve tissues and provide acceptable clinical removal rates. Spectral analysis of the different tooth tissues is also conducted and differences between the visible wavelength fsLIBS spectra are evident, though more robust classification studies are needed for clinical translation. These results have initiated a set of precautionary recommendations that would enable the clinician to utilize femtosecond laser ablation for the removal of carious lesions while ensuring that the solidity and utility of the tooth remain intact. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Experimental investigation on the caries characteristic of dental tissues by photothermal radiometry scanning imaging

    NASA Astrophysics Data System (ADS)

    Wang, Fei; Liu, Jun-yan; Wang, Xiao-chun; Wang, Yang

    2018-03-01

    In this paper, a one-dimensional (1D) thermal-wave model coupled diffuse-photon-density-wave for three-layer dental tissues using modulated laser stimulation was employed to illustrate the relationship between dental caries characteristic (i.e. caries layer thickness, optical absorption coefficient and optical scattering coefficient) and photothermal radiometry (PTR) signal. Experimental investigation of artificial caries was carried out using PTR scanning imaging. The PTR amplitude and phase delay were increased with dental demineralized treatment. The local caries characteristic parameters were obtained by the best-fitting method based on the 1D thermal-wave model. The PTR scanning imaging measurements illustrated that the optical absorption coefficient and scattering coefficient of caries region were much higher than those of the healthy enamel area. The demineralization thickness of caries region was measured by PTR scanning imaging and its average value shows in good agreement with the digital microscope. Experimental results show that PTR scanning imaging has the merits of high contrast for local inhomogeneity of dental caries; furthermore, this method is an allowance to provide a flexibility for non-contact quantitative evaluation of dental caries.

  17. Infrared spectroscopy of molecular submonolayers on surfaces by infrared scanning tunneling microscopy: tetramantane on Au111.

    PubMed

    Pechenezhskiy, Ivan V; Hong, Xiaoping; Nguyen, Giang D; Dahl, Jeremy E P; Carlson, Robert M K; Wang, Feng; Crommie, Michael F

    2013-09-20

    We have developed a new scanning-tunneling-microscopy-based spectroscopy technique to characterize infrared (IR) absorption of submonolayers of molecules on conducting crystals. The technique employs a scanning tunneling microscope as a precise detector to measure the expansion of a molecule-decorated crystal that is irradiated by IR light from a tunable laser source. Using this technique, we obtain the IR absorption spectra of [121]tetramantane and [123]tetramantane on Au(111). Significant differences between the IR spectra for these two isomers show the power of this new technique to differentiate chemical structures even when single-molecule-resolved scanning tunneling microscopy (STM) images look quite similar. Furthermore, the new technique was found to yield significantly better spectral resolution than STM-based inelastic electron tunneling spectroscopy, and to allow determination of optical absorption cross sections. Compared to IR spectroscopy of bulk tetramantane powders, infrared scanning tunneling microscopy (IRSTM) spectra reveal narrower and blueshifted vibrational peaks for an ordered tetramantane adlayer. Differences between bulk and surface tetramantane vibrational spectra are explained via molecule-molecule interactions.

  18. Globular domain of adiponectin: promising target molecule for detection of atherosclerotic lesions

    PubMed Central

    Almer, Gunter; Saba-Lepek, Matthias; Haj-Yahya, Samih; Rohde, Eva; Strunk, Dirk; Fröhlich, Eleonore; Prassl, Ruth; Mangge, Harald

    2011-01-01

    Background: Adiponectin, an adipocyte-specific plasma protein, has been shown to accumulate in injured endothelial cells during development of atherosclerotic lesions. In this study, we investigated the potential of different adiponectin subfractions with special emphasis on globular adiponectin (gAd) to recognize and visualize atherosclerotic lesions. Methods: Recombinant mouse gAd and subfractions of full-length adiponectin (ie, trimeric, hexameric, and oligomeric forms) were fluorescence-labeled. Aortas of wild-type and apoprotein E-deficient mice fed a high cholesterol diet were dissected and incubated with the labeled biomarkers. Imaging was performed using confocal laser scanning microscopy. Results: Confocal laser scanning microscopic images showed that gAd binds more strongly to atherosclerotic plaques than full-length adiponectin subfractions. Further, we showed that gAd accumulates preferentially in endothelial cells and the fibrous cap area of plaques. Here we demonstrate for the first time that gAd recognizes atherosclerotic plaques on aortic sections of apoprotein E-deficient mice. Conclusion: These results suggest that gAd, in addition to its physiological properties, is also suitable as a target molecule for prospective diagnostic strategies in imaging atherosclerotic lesions. PMID:22022204

  19. A laser scanning confocal imaging-surface plasmon resonance system application in real time detection of antibody-antigen interaction

    NASA Astrophysics Data System (ADS)

    Zhang, H. Y.; Yang, L. Q.; Liu, W. M.

    2011-12-01

    The laser scanning confocal microscope (LSCM) offers several advantages over conventional optical microscopy, but most LSCM work is qualitative analysis and it is very hard to achieve quantitative detection directly with the changing of the fluorescent intensity. A new real time sensor system for the antibody-antigen interaction detection was built integrating with a LSCM and a wavelength-dependent surface plasmon resonance (SPR) sensor. The system was applied to detect the bonding process of human IgG and fluorescent-labeled affinity purified antibody in real time. The fluorescence images changing is well with that of SPR wavelengths in real time, and the trend of the resonance wavelength shift with the concentrations of antibody is similar to that of the fluorescent intensity changing. The results show that SPR makes up the short of quantificational analysis with LSCM with the high spatial resolution. The sensor system shows the merits of the of the LSCM and SPR synergetic application, which are of great importance for practical application in biosensor and life science for interesting local interaction.

  20. Laser Scanning Cytometry

    PubMed Central

    Pozarowski, Piotr; Holden, Elena; Darzynkiewicz, Zbigniew

    2013-01-01

    Summary The laser scanning cytometer (LSC) is the microscope-based cytofluorometer that offers a plethora of analytical capabilities. Multilaser-excited fluorescence emitted from individual cells is measured at several wavelength ranges, rapidly (up to 5000 cells/min), with high sensitivity and accuracy. The following applications of LSC are reviewed: (1) identification of cells that differ in degree of chromatin condensation (e.g., mitotic or apoptotic cells or lymphocytes vs granulocytes vs monocytes); (2) detection of translocation between cytoplasm vs nucleus or nucleoplasm vs nucleolus of regulatory molecules such as NF- κB, p53, or Bax; (3) semiautomatic scoring of micronuclei in mutagenicity assays; (4) analysis of fluorescence in situ hybridization; (5) enumeration and morphometry of nucleoli; (6) analysis of phenotype of progeny of individual cells in clonogenicity assay; (7) cell immunophenotyping; (8) visual examination, imaging, or sequential analysis of the cells measured earlier upon their relocation, using different probes; (9) in situ enzyme kinetics and other time-resolved processes; (10) analysis of tissue section architecture; (11) application for hypocellular samples (needle aspirate, spinal fluid, etc.); (12) other clinical applications. Advantages and limitations of LSC are discussed and compared with flow cytometry. PMID:16719355

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