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Sample records for scanning laser confocal

  1. QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY

    Directory of Open Access Journals (Sweden)

    Merete Krog Raarup

    2011-05-01

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

  2. Application of Confocal Laser Scanning Microscopy in Biology and Medicine

    OpenAIRE

    I. A. Volkov; N. V. Frigo; L. F. Znamenskaya; O. R. Katunina

    2014-01-01

    Fluorescence confocal laser scanning microscopy and reflectance confocal laser scanning microscopy are up-to-date highend study methods. Confocal microscopy is used in cell biology and medicine. By using confocal microscopy, it is possible to study bioplasts and localization of protein molecules and other compounds relative to cell or tissue structures, and to monitor dynamic cell processes. Confocal microscopes enable layer-by-layer scanning of test items to create demonstrable 3D models. As...

  3. A New Multichannel Spectral Imaging Laser Scanning Confocal Microscope

    Directory of Open Access Journals (Sweden)

    Yunhai Zhang

    2013-01-01

    Full Text Available We have developed a new multichannel spectral imaging laser scanning confocal microscope for effective detection of multiple fluorescent labeling in the research of biological tissues. In this paper, the design and key technologies of the system are introduced. Representative results on confocal imaging, 3-dimensional sectioning imaging, and spectral imaging are demonstrated. The results indicated that the system is applicable to multiple fluorescent labeling in biological experiments.

  4. Optomechatronics Design and Control for Confocal Laser Scanning Microscopy

    NARCIS (Netherlands)

    Yoo, H.W.

    2015-01-01

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

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

    Science.gov (United States)

    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.

  6. Signal and noise modeling in confocal laser scanning fluorescence microscopy.

    Science.gov (United States)

    Herberich, Gerlind; Windoffer, Reinhard; Leube, Rudolf E; Aach, Til

    2012-01-01

    Fluorescence confocal laser scanning microscopy (CLSM) has revolutionized imaging of subcellular structures in biomedical research by enabling the acquisition of 3D time-series of fluorescently-tagged proteins in living cells, hence forming the basis for an automated quantification of their morphological and dynamic characteristics. Due to the inherently weak fluorescence, CLSM images exhibit a low SNR. We present a novel model for the transfer of signal and noise in CLSM that is both theoretically sound as well as corroborated by a rigorous analysis of the pixel intensity statistics via measurement of the 3D noise power spectra, signal-dependence and distribution. Our model provides a better fit to the data than previously proposed models. Further, it forms the basis for (i) the simulation of the CLSM imaging process indispensable for the quantitative evaluation of CLSM image analysis algorithms, (ii) the application of Poisson denoising algorithms and (iii) the reconstruction of the fluorescence signal.

  7. Confocal laser scanning microscopy in study of bone calcification

    Science.gov (United States)

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

    2012-12-01

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

  8. Confocal laser scanning microscopy in study of bone calcification

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-12-01

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

  9. Confocal laser scanning microscopy in study of bone calcification

    International Nuclear Information System (INIS)

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

    2012-01-01

    Highlights: ► High-magnification images with depth selection, and thin sections were observed using CLSM. ► The direction and velocity of calcification of the bone was observed by administration of 2 fluorescent dyes. ► In dog femora grafted with coral blocks, newly-formed bone was observed in the coral block space with a rough surface. ► Twelve weeks after dental implant was grafted in dog femora, the space between screws was filled with newly-formed bones. - Abstract: Bone regeneration in mandible and maxillae after extraction of teeth or tumor resection and the use of rough surface implants in bone induction must be investigated to elucidate the mechanism of calcification. The calcified tissues are subjected to chemical decalcification or physical grinding to observe their microscopic features with light microscopy and transmission electron microscopy where the microscopic tissue morphology is significantly altered. We investigated the usefulness of confocal laser scanning microscopy (CLSM) for this purpose. After staggering the time of administration of calcein and alizarin red to experimental rats and dogs, rat alveolar bone and dog femur grafted with coral as scaffold or dental implants were observed with CLSM. In rat alveolar bone, the calcification of newly-formed bone and net-like canaliculi was observed at the mesial bone from the roots progressed at the rate of 15 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.

  10. Visualization of carbon nanotubes dispersion in composite by using confocal laser scanning microscopy

    Czech Academy of Sciences Publication Activity Database

    Ilčíková, M.; Danko, M.; Doroshenko, M.; Best, A.; Mrlík, M.; Csomorová, K.; Šlouf, Miroslav; Chorvát Jr., D.; Koynov, K.; Mosnáček, J.

    2016-01-01

    Roč. 79, June (2016), s. 187-197 ISSN 0014-3057 Institutional support: RVO:61389013 Keywords : confocal laser scanning microscopy * composites * carbon nanotubes dispersion Subject RIV: CD - Macromolecular Chemistry Impact factor: 3.531, year: 2016

  11. Clinical applications of in vivo fluorescence confocal laser scanning microscopy

    Science.gov (United States)

    Oh, Chilhwan; Park, Sangyong; Kim, Junhyung; Ha, Seunghan; Park, Gyuman; Lee, Gunwoo; Lee, Onseok; Chun, Byungseon; Gweon, Daegab

    2008-02-01

    Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively. CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin. In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens, 0.9NA oil immersion, axial resolution 1.0μm, field of view 200μm x 100μm (lateral resolution , 0.3μm). In human volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. In

  12. Transient gels in colloid-polymer mixtures studied with fluorescence confocal scanning laser microscopy

    NARCIS (Netherlands)

    Verhaegh, N.A.M.; Asnaghi, D.; Lekkerkerker, H.N.W.

    1999-01-01

    We study the structure and the time evolution of transient gels formed in colloid-polymer mixtures, by means of uorescence Confocal Scanning Laser Microscopy (CSLM). This technique is used in conjunction with novel colloidal silica particles containing a uorescent core. The confocal micrographs

  13. A confocal scanning laser ophthalmoscope for retinal vessel oximetry

    Science.gov (United States)

    Lompado, Arthur

    Measurement of a person's blood oxygen saturation has long been recognized as a useful metric for the characterizing ailments ranging from chronic respiratory disorders to acute, potentially life threatening, traumas. The ubiquity of oxygen saturation monitors in the medical field, including portable pulse oximeters and laboratory based CO-oximeters, is a testament to the importance of this technique. The work presented here documents the design, fabrication and development of a unique type of oxygen saturation monitor, a confocal scanning retinal vessel oximeter, with the potential to expand the usefulness of the present devices. A large part of the knowledge base required to construct the instrument comes from the consideration of light scattering by red blood cells in a blood vessel. Therefore, a substantial portion of this work is devoted to the process of light scattering by whole human blood and its effects on the development of a more accurate oximeter. This light scattering effect has been both measured and modeled stochastically to determine its contribution to the measured oximeter signal. It is shown that, although well accepted in the published literature, the model only correlates marginally to the measurements due to inherent limitations imposed by the model assumptions. Nonetheless, enough material has been learned about the scattering to allow development of a mathematical model for the interaction of light with blood in a vessel, and this knowledge has been applied to the data reduction of the present oximeter. This data reduction technique has been tested in a controlled experiment employing a model eye with a blood filled mock retinal vessel. It will be shown that the presently developed technique exhibited strong correlation between the known blood oxygen saturation and that calculated by the new system.

  14. Musculature of Notholca acuminata (Rotifera : Ploima : Brachionidae) revealed by confocal scanning laser microscopy

    DEFF Research Database (Denmark)

    Sørensen, M.V.; Funch, P.; Hooge, M.

    2003-01-01

    The body-wall and visceral musculature of Notholca acuminata was visualized using phalloidin-linked fluorescent dye under confocal laser scanning microscopy. The body-wall musculature includes dorsal, lateral, and ventral pairs of longitudinally oriented body retractor muscles, two pairs of head...

  15. Application of the laser scanning confocal microscope in fluorescent film sensor research

    Science.gov (United States)

    Zhang, Hongyan; Liu, Wei-Min; Zhao, Wen-Wen; Dai, Qing; Wang, Peng-Fei

    2010-10-01

    Confocal microscopy offers several advantages over conventional optical microscopy; we show an experimental investigation laser scanning confocal microscope as a tool to be used in cubic boron nitride (cBN) film-based fluorescent sensor research. Cubic boron nitride cBN film sensors are modified with dansyl chloride and rhodamine B isothiocyanate respectively. Fluorescent modification quality on the cubic boron nitride film is clearly express and the sensor ability to Hg2+ cations and pH are investigated in detail. We evidence the rhodamine B isothiocyanate modified quality on cBN surface is much better than that of dansyl chloride. And laser scanning confocal microscope has potential application lighttight fundus film fluorescent sensor research.

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

    Science.gov (United States)

    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.

  17. Mechanisms of biliary stent clogging: confocal laser scanning and scanning electron microscopy.

    Science.gov (United States)

    van Berkel, A M; van Marle, J; Groen, A K; Bruno, M J

    2005-08-01

    Endoscopic insertion of plastic biliary endoprostheses is a well-established treatment for obstructive jaundice. The major limitation of this technique is late stent occlusion. In order to compare events involved in biliary stent clogging and identify the distribution of bacteria in unblocked stents, confocal laser scanning (CLS) and scanning electron microscopy (SEM) were carried out on two different stent materials - polyethylene (PE) and hydrophilic polymer-coated polyurethane (HCPC). Ten consecutive patients with postoperative benign biliary strictures were included in the study. Two 10-Fr stents 9 cm in length, one made of PE and the other of HCPC, were inserted. The stents were electively exchanged after 3 months and examined using CLS and SEM. No differences were seen between the two types of stent. The inner stent surface was covered with a uniform amorphous layer. On top of this layer, a biofilm of living and dead bacteria was found, which in most cases was unstructured. The lumen was filled with free-floating colonies of bacteria and crystals, surrounded by mobile laminar structures of mucus. An open network of large dietary fibers was seen in all of the stents. The same clogging events occurred in both PE and HCPC stents. The most remarkable observation was the identification of networks of large dietary fibers, resulting from duodenal reflux, acting as a filter. The build-up of this intraluminal framework of dietary fibers appears to be a major factor contributing to the multifactorial process of stent clogging.

  18. Methods for studying biofilm formation: flow cells and confocal laser scanning microscopy

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2014-01-01

    In this chapter methods for growing and analyzing biofilms under hydrodynamic conditions in flow cells are described. Use of flow cells allows for direct microscopic investigation of biofilm formation. The flow in these chambers is essentially laminar, which means that the biofilms can be grown u......, inoculation of the flow cells, running of the system, confocal laser scanning microscopy and image analysis, and disassembly and cleaning of the system....

  19. Atomic force microscopy and confocal laser scanning microscopy on the cytoskeleton of permeabilised and embedded cells

    International Nuclear Information System (INIS)

    Meller, Karl; Theiss, Carsten

    2006-01-01

    We describe a technical method of cell permeabilisation and embedding to study the organisation and distribution of intracellular proteins with aid of atomic force microscopy and confocal laser scanning microscopy in identical areas. While confocal laser scanning microscopy is useful for the identification of certain proteins subsequent labelling with markers or antibodies, atomic force microscopy allows the observation of macromolecular structures in fixed and living cells. To demonstrate the field of application of this preparatory technique, cells were permeabilised, fixed, and the actin cytoskeleton was stained with phalloidin-rhodamine. Confocal laser scanning microscopy was used to show the organisation of these microfilaments, e.g. geodesic dome structures. Thereafter, cells were embedded in Durcupan water-soluble resin, followed by UV-polymerisation of resin at 4 o C. This procedure allowed intracellular visualisation of the cell nucleus or cytoskeletal elements by atomic force microscopy, for instance to analyse the globular organisation of actin filaments. Therefore, this method offers a great potential to combine both microscopy techniques in order to understand and interpret intracellular protein relations, for example, the biochemical and morphological interaction of the cytoskeleton

  20. 3-D reconstruction of neurons from multichannel confocal laser scanning image series.

    Science.gov (United States)

    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.

  1. Optical coherence tomography, scanning laser polarimetry and confocal scanning laser ophthalmoscopy in retinal nerve fiber layer measurements of glaucoma patients.

    Science.gov (United States)

    Fanihagh, Farsad; Kremmer, Stephan; Anastassiou, Gerasimos; Schallenberg, Maurice

    2015-01-01

    To determine the correlations and strength of association between different imaging systems in analyzing the retinal nerve fiber layer (RNFL) of glaucoma patients: optical coherence tomography (OCT), scanning laser polarimetry (SLP) and confocal scanning laser ophthalmoscopy (CSLO). 114 eyes of patients with moderate open angle glaucoma underwent spectral domain OCT (Topcon SD-OCT 2000 and Zeiss Cirrus HD-OCT), SLP (GDx VCC and GDx Pro) and CSLO (Heidelberg Retina Tomograph, HRT 3). Correlation coefficients were calculated between the structural parameters yielded by these examinations. The quantitative relationship between the measured RNFL thickness globally and for the four regions (superior, inferior, nasal, temporal) were evaluated with different regression models for all used imaging systems. The strongest correlation of RNFL measurements was found between devices using the same technology like GDx VCC and GDx Pro as well as Topcon OCT and Cirrus OCT. In glaucoma patients, the strongest associations (R²) were found between RNFL measurements of the two optical coherence tomography devices Topcon OCT and Cirrus OCT (R² = 0.513) and between GDx VCC and GDx Pro (R² = 0.451). The results of the OCTs and GDX Pro also had a strong quantitative relationship (Topcon OCT R² = 0.339 and Cirrus OCT R² = 0.347). GDx VCC and the OCTs showed a mild to moderate association (Topcon OCT R² = 0.207 and Cirrus OCT R² = 0.258). The confocal scanning laser ophthalmoscopy (HRT 3) had the lowest association to all other devices (Topcon OCT R² = 0.254, Cirrus OCT R² = 0.158, GDx Pro R² = 0.086 and GDx VCC R² = 0.1). The measurements of the RNFL in glaucoma patients reveal a high correlation of OCT and GDx devices because OCTs can measure all major retinal layers and SLP can detect nerve fibers allowing a comparison between the results of this devices. However, CSLO by means of HRT topography can only measure height values of the retinal surface but it cannot distinguish

  2. Investigation of phosphatidylcholine enhancing FITC-insulin across buccal mucosa by confocal laser scanning microscopy

    Science.gov (United States)

    Tian, Weiqun; Su, Li; Zeng, Shaoqun; Luo, Qingming; Gao, Qiuhua; Xu, Huibi

    2002-04-01

    The aim was to characterize the transport of fluorescein isothiocyanate (FITC)-labeled dextran and insulin with different resoluble compounds for peptides and proteins through buccal mucosa. The penetration rate of insulin molecules through porcine buccal mucosa (a nonkeratinized epithelium, comparable to human buccal mucosa) was investigated by measuring transbuccal fluxes and by analyzing the distribution of the fluorescent probe in the rabbit buccal mucosa epithelium, using confocal laser scanning microscopy for visualizing permeation pathways. The confocal images of the distribution pattern of FITC-dextran and FITC-insulin showed that the paracellular route is the major pathway of FITC-dextran through buccal mucosa epithelium, the intra-cellular route is the major pathway of FITC-insulin through buccal mucosa epithelium. The permeation rate can be increased by co-administration of soybean phosphatidylcholine (SPC).

  3. Confocal laser scanning microscopy to estimate nanoparticles’ human skin penetration in vitro

    Directory of Open Access Journals (Sweden)

    Zou Y

    2017-10-01

    Full Text Available Ying Zou,1,2,* Anna Celli,2,3,* Hanjiang Zhu,2,* Akram Elmahdy,2 Yachao Cao,2 Xiaoying Hui,2 Howard Maibach2 1Skin & Cosmetic Research Department, Shanghai Skin Disease Hospital, Shanghai, People’s Republic of China; 2Department of Dermatology, School of Medicine, University of California San Francisco, San Francisco, CA, USA; 3San Francisco Veterans Medical Center, San Francisco, CA, USA *These authors contributed equally to this work Objective: With rapid development of nanotechnology, there is increasing interest in nanoparticle (NP application and its safety and efficacy on human skin. In this study, we utilized confocal laser scanning microscopy to estimate NP skin penetration.Methods: Three different-sized polystyrene NPs marked with red fluorescence were applied to human skin, and Calcium Green 5N was used as a counterstain. Dimethyl sulfoxide (DMSO and ethanol were used as alternative vehicles for NPs. Tape stripping was utilized as a barrier-damaged skin model. Skin biopsies dosed with NPs were incubated at 4°C or 37°C for 24 hours and imaged using confocal laser scanning microscopy.Results: NPs were localized in the stratum corneum (SC and hair follicles without penetrating the epidermis/dermis. Barrier alteration with tape stripping and change in incubation temperature did not induce deeper penetration. DMSO enhanced NP SC penetration but ethanol did not.Conclusion: Except with DMSO vehicle, these hydrolyzed polystyrene NPs did not penetrate intact or barrier-damaged human “viable” epidermis. For further clinical relevance, in vivo human skin studies and more sensitive analytic chemical methodology are suggested. Keywords: nanoparticles, skin penetration, stratum corneum, confocal laser scanning microscopy, tape stripping

  4. Three-dimensional imaging of porous media using confocal laser scanning microscopy.

    Science.gov (United States)

    Shah, S M; Crawshaw, J P; Boek, E S

    2017-02-01

    In the last decade, imaging techniques capable of reconstructing three-dimensional (3-D) pore-scale model have played a pivotal role in the study of fluid flow through complex porous media. In this study, we present advances in the application of confocal laser scanning microscopy (CLSM) to image, reconstruct and characterize complex porous geological materials with hydrocarbon reservoir and CO 2 storage potential. CLSM has a unique capability of producing 3-D thin optical sections of a material, with a wide field of view and submicron resolution in the lateral and axial planes. However, CLSM is limited in the depth (z-dimension) that can be imaged in porous materials. In this study, we introduce a 'grind and slice' technique to overcome this limitation. We discuss the practical and technical aspects of the confocal imaging technique with application to complex rock samples including Mt. Gambier and Ketton carbonates. We then describe the complete workflow of image processing to filtering and segmenting the raw 3-D confocal volumetric data into pores and grains. Finally, we use the resulting 3-D pore-scale binarized confocal data obtained to quantitatively determine petrophysical pore-scale properties such as total porosity, macro- and microporosity and single-phase permeability using lattice Boltzmann (LB) simulations, validated by experiments. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

  5. Confocal laser scanning microscopy in vivo for diagnosing melanocytic skin neoplasms

    Directory of Open Access Journals (Sweden)

    A. A. Kubanova

    2014-01-01

    Full Text Available The authors discuss the use of confocal laser scanning microscopy in vivo (CLSM for diagnosing melanocytic skin neoplasms and its value for early diagnostics of melanoma. CLSM is an innovation noninvasive visual examination method for real-time multiple and painless examinations of the patient’s skin without injuring the skin integument. The method ensures early diagnostics of skin melanomas with high sensitivity and specificity, which makes it possible to use CLSM for screening melanocytic skin neoplasms for the sake of the early onset of treatment to save patient life and health.

  6. Ti-6Al-4V electron beam weld qualification using laser scanning confocal microscopy

    International Nuclear Information System (INIS)

    Wanjara, P.; Brochu, M.; Jahazi, M.

    2005-01-01

    Processing conditions for manufacturing Ti-6Al-4V components by welding using an electron beam source are known to influence the transformation microstructure in the narrow fusion and heat-affected zones of the weld region. This work examined the effect of multiple-sequence welding on the characteristics of the transformed beta microstructure, using laser scanning confocal microscopy to resolve the Widmanstaetten alpha-beta structure in the fusion zone. The evolution in the alpha interlamellar spacing and plate thickness with processing was then related to microhardness measurements in the weld region

  7. Embryological study of Herminium monorchis (Orchidaceae) using confocal scanning laser microscopy

    International Nuclear Information System (INIS)

    Fredrikson, M.

    1990-01-01

    The embryology of Herminium monorchis (Orchidaceae) was studied using confocal scanning laser microscopy (CSLM), a new technique for embryological studies. This technique may contribute new information to plant embryology. Herminium monorchis has a monosporic embryo sac development. The mature embryo sac is 8-nucleate. Two integuments, both 2-layered, are formed, but only the inner takes part in formation of the micropyle. Double fertilization takes place. The primary endosperm nucleus does not divide, but remains alive at least at the 3-celled stage of embryo development. The three antipodals do not show any sign of degeneration at this stage. (author)

  8. Characterization of particle deformation during compression measured by confocal laser scanning microscopy.

    Science.gov (United States)

    Guo, H X; Heinämäki, J; Yliruusi, J

    1999-09-20

    Direct compression of riboflavin sodium phosphate tablets was studied by confocal laser scanning microscopy (CLSM). The technique is non-invasive and generates three-dimensional (3D) images. Tablets of 1% riboflavin sodium phosphate with two grades of microcrystalline cellulose (MCC) were individually compressed at compression forces of 1.0 and 26.8 kN. The behaviour and deformation of drug particles on the upper and lower surfaces of the tablets were studied under compression forces. Even at the lower compression force, distinct recrystallized areas in the riboflavin sodium phosphate particles were observed in both Avicel PH-101 and Avicel PH-102 tablets. At the higher compression force, the recrystallization of riboflavin sodium phosphate was more extensive on the upper surface of the Avicel PH-102 tablet than the Avicel PH-101 tablet. The plastic deformation properties of both MCC grades reduced the fragmentation of riboflavin sodium phosphate particles. When compressed with MCC, riboflavin sodium phosphate behaved as a plastic material. The riboflavin sodium phosphate particles were more tightly bound on the upper surface of the tablet than on the lower surface, and this could also be clearly distinguished by CLSM. Drug deformation could not be visualized by other techniques. Confocal laser scanning microscopy provides valuable information on the internal mechanisms of direct compression of tablets.

  9. Automatic segmentation of cell nuclei from confocal laser scanning microscopy images

    International Nuclear Information System (INIS)

    Kelemen, A.; Reist, H.W.

    1997-01-01

    A newly developed experimental method combines the possibility of irradiating more than a thousand cells simultaneous with an efficient colony-forming ability and with the capability of localizing a particle track through a cell nucleus together with the assessment of the energy transfer by digital superposition of the image containing the track with that of the cells. To assess the amount of energy deposition by particles traversing the cell nucleus the intersection lengths of the particle tracks have to be known. Intersection lengths can be obtained by determining the 3D surface contours of the irradiated cell nuclei. Confocal laser scanning microscopy using specific DNA fluorescent dye offers a possible way for the determination of the 3D shape of individual nuclei. Unfortunately, such experiments cannot be performed on living cells. One solution to this problem can be provided by building a statistical model of the shape of the nuclei of the exposed cells. In order to build such a statistical model, a large number of cell nuclei have to be identified and segmented from confocal laser scanning microscopy images. The present paper describes a method to perform this 3D segmentation in an automatic manner in order to create a solid basis for the statistical model. (author) 2 figs., 4 refs

  10. Confocal laser scanning microscopy to estimate nanoparticles' human skin penetration in vitro.

    Science.gov (United States)

    Zou, Ying; Celli, Anna; Zhu, Hanjiang; Elmahdy, Akram; Cao, Yachao; Hui, Xiaoying; Maibach, Howard

    2017-01-01

    With rapid development of nanotechnology, there is increasing interest in nanoparticle (NP) application and its safety and efficacy on human skin. In this study, we utilized confocal laser scanning microscopy to estimate NP skin penetration. Three different-sized polystyrene NPs marked with red fluorescence were applied to human skin, and Calcium Green 5N was used as a counterstain. Dimethyl sulfoxide (DMSO) and ethanol were used as alternative vehicles for NPs. Tape stripping was utilized as a barrier-damaged skin model. Skin biopsies dosed with NPs were incubated at 4°C or 37°C for 24 hours and imaged using confocal laser scanning microscopy. NPs were localized in the stratum corneum (SC) and hair follicles without penetrating the epidermis/dermis. Barrier alteration with tape stripping and change in incubation temperature did not induce deeper penetration. DMSO enhanced NP SC penetration but ethanol did not. Except with DMSO vehicle, these hydrolyzed polystyrene NPs did not penetrate intact or barrier-damaged human "viable" epidermis. For further clinical relevance, in vivo human skin studies and more sensitive analytic chemical methodology are suggested.

  11. Confocal laser scanning microscopy to estimate nanoparticles’ human skin penetration in vitro

    Science.gov (United States)

    Elmahdy, Akram; Cao, Yachao; Hui, Xiaoying; Maibach, Howard

    2017-01-01

    Objective With rapid development of nanotechnology, there is increasing interest in nanoparticle (NP) application and its safety and efficacy on human skin. In this study, we utilized confocal laser scanning microscopy to estimate NP skin penetration. Methods Three different-sized polystyrene NPs marked with red fluorescence were applied to human skin, and Calcium Green 5N was used as a counterstain. Dimethyl sulfoxide (DMSO) and ethanol were used as alternative vehicles for NPs. Tape stripping was utilized as a barrier-damaged skin model. Skin biopsies dosed with NPs were incubated at 4°C or 37°C for 24 hours and imaged using confocal laser scanning microscopy. Results NPs were localized in the stratum corneum (SC) and hair follicles without penetrating the epidermis/dermis. Barrier alteration with tape stripping and change in incubation temperature did not induce deeper penetration. DMSO enhanced NP SC penetration but ethanol did not. Conclusion Except with DMSO vehicle, these hydrolyzed polystyrene NPs did not penetrate intact or barrier-damaged human “viable” epidermis. For further clinical relevance, in vivo human skin studies and more sensitive analytic chemical methodology are suggested. PMID:29184403

  12. QUANTIFICATION OF BIOFILMS IN MULTI-SPECTRAL DIGITAL1 VOLUMES FROM CONFOCAL LASER-SCANNING MICROSCOPES

    Directory of Open Access Journals (Sweden)

    Karsten Rodenacker

    2011-05-01

    Full Text Available Populations of bacteria in sludge flocs and biofilm marked by fluorescence marked with fluorescent probes are digitised with a confocal laser scanning microscope. These data are used to analyse the microbial community structure, to obtain information on the localisation of specific bacterial groups and to examine gene expression. This information is urgently required for an in-depth understanding of the function and, more generally, the microbial ecology of biofilms. Methods derived from quantitative image analysis are applied to digitised data from confocal laser scanning microscopes to obtain quantitative descriptions of volumetric, topological (and topographical properties of different compartments of the components under research. In addition to free-moving flocs, also biofilms attached to a substratum in an experimental environment are analysed. Growth form as well as interaction of components are quantitatively described. Classical measurements of volume and intensity (shape, distribution and distance dependent interaction measurements using methods from mathematical morphology are performed. Mainly image (volume processing methods are outlined. Segmented volumes are globally and individually (in terms of 3Dconnected components measured and used for distance mapping transform as well as for estimation of geodesic distances from the substrate. All transformations are applied on the 3D data set. Resulting distance distributions are quantified and related to information on the identity and activity of the probe-identified bacteria.

  13. Iris ultrastructure in patients with synechiae as revealed by in vivo laser scanning confocal microscopy : In vivo iris ultrastructure in patients with Synechiae by Laser Scanning Confocal Microscopy.

    Science.gov (United States)

    Li, Ming; Cheng, Hongbo; Guo, Ping; Zhang, Chun; Tang, Song; Wang, Shusheng

    2016-04-26

    Iris plays important roles in ocular physiology and disease pathogenesis. Currently it is technically challenging to noninvasively examine the human iris ultrastructure in vivo. The purpose of the current study is to reveal human iris ultrastructure in patients with synechiae by using noninvasive in vivo laser scanning confocal microscopy (LSCM). The ultrastructure of iris in thirty one patients, each with synechiae but transparent cornea, was examined by in vivo LSCM. Five characteristic iris ultrastructures was revealed in patients with synechiae by in vivo LSCM, which include: 1. tree trunk-like structure; 2. tree branch/bush-like structure; 3. Fruit-like structure; 4. Epithelioid-like structure; 5. deep structure. Pigment granules can be observed as a loose structure on the top of the arborization structure. In iris-associated diseases with Tyndall's Phenomenon and keratic precipitates, the pigment particles are more likely to fall off from the arborization structure. The ultrastructure of iris in patients with synechiae has been visualized using in vivo LSCM. Five iris ultrastructures can be clearly observed, with some of the structures maybe disease-associated. The fall-off of the pigment particles may cause the Tyndall's Phenomenon positive. In vivo LSCM provides a non-invasive approach to observe the human iris ultrastructure under certain eye disease conditions, which sets up a foundation to visualize certain iris-associated diseases in the future.

  14. Investigation of the petrophysical properties of a porous sandstone sample using confocal scanning laser microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Petford, N. [Kingston Univ., Centre for Earth and Environmental Science Research, Kingston (United Kingdom); Davidson, G. [University Coll., Dept. of Electronic and Electrical Engineering, London (United Kingdom); Miller, J.A. [Cambridge Univ., Dept. of Earth Sciences, Cambridge (United Kingdom)

    2001-05-01

    Confocal scanning laser microscopy (CSLM) is used to produce images of the two- and three-dimensional distribution and geometry of pore space in a reservoir sandstone and measure the 2D distribution of pore throat radii. Non-destructive serial sectioning of the rock using laser light at 100% illumination, combined with image thresholding and histogram equalization techniques allow the pore volume structure of the uppermost 100 {mu}m of the sample to be reconstructed. Negative imaging of the pore volume gave superior depth and feature resolution compared to positive (reflection) imaging. Artefacts encountered in applying classical Medial Axial Transforms to CSLM images include branch networks dominated by coordination numbers of 3. Skeletonization using Euclidean distance maps gives increased accuracy in the description of the pore network. Measured pore throat size distribution in the rock is strongly exponential and described by the expression y 219e{sup -0.25x} where y is the number of pore throats. (Author)

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    The microstructure of protein networks in yogurts defines important physical properties of the yogurt and hereby partly its quality. Imaging this protein network using confocal scanning laser microscopy (CSLM) has shown good results, and CSLM has become a standard measuring technique for fermented...... to image texture description. Here, CSLM images from a yogurt fermentation study are investigated, where production factors including fat content, protein content, heat treatment, and incubation temperature are varied. The descriptors are evaluated through nearest neighbor classification, variance analysis...... scanning microscopy images can be used to provide information on the protein microstructure in yogurt products. For large numbers of microscopy images, subjective evaluation becomes a difficult or even impossible approach, if the images should be incorporated in any form of statistical analysis alongside...

  16. Analysis of polymer grafted inside the porous hydrogel using confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    2007-04-01

    Full Text Available Graft polymerization of glycidyl methacrylate onto the pore surface of polyacrylamide macroporous gel was implemented in DMSO-aqueous solution using diperiodatocuprate(III complexes as an initiator. The grafting densities up to 410% were achieved. The graft polymerization was confirmed by gravimetrical methods and FTIR. The graft polymerization of polymer inside the pores of the macroporous gel resulted in increased flow resistance through the gel matrix. The distribution of grafted polymer on the gel pore surface material was studied by scanning electron microscopy (SEM and confocal laser scanning microscopy (CLSM. CLSM is an alternative method for studying morphology of gel surface with grafted polymer having the advantages over the SEM allowing to investigate the distribution of grafted polymer inside the hydrogel in a native hydrated state. The microscopic techniques demonstrated uneven distribution of the grafted polymer inside the gel pores as a result of initiating the graft polymerization by insoluble initiator deposited on the pore surface.

  17. Confocal laser scanning microscopy detection of chlorophylls and carotenoids in chloroplasts and chromoplasts of tomato fruit.

    Science.gov (United States)

    D'Andrea, Lucio; Amenós, Montse; Rodríguez-Concepción, Manuel

    2014-01-01

    Plant cells are unique among eukaryotic cells because of the presence of plastids, including chloroplasts and chromoplasts. Chloroplasts are found in green tissues and harbor the photosynthetic machinery (including chlorophyll molecules), while chromoplasts are present in non-photosynthetic tissues and accumulate large amounts of carotenoids. During tomato fruit development, chloroplasts are converted into chromoplasts that accumulate high levels of lycopene, a linear carotenoid responsible for the characteristic red color of ripe fruit. Here, we describe a simple and fast method to detect both types of fully differentiated plastids (chloroplasts and chromoplasts), as well as intermediate stages, in fresh tomato fruits. The method is based on the differential autofluorescence of chlorophylls and carotenoids (lycopene) detected by Confocal Laser Scanning Microscopy.

  18. Spectral imaging technique for retinal perfusion detection using confocal scanning laser ophthalmoscopy

    Science.gov (United States)

    Rasta, Seyed Hossein; Manivannan, Ayyakkannu; Sharp, Peter F.

    2012-11-01

    To evaluate retinal perfusion in the human eye, a dual-wavelength confocal scanning laser ophthalmoscope (cSLO) was developed that provides spectral imaging of the fundus using a combination of red (670 nm) and near-infrared (810 nm) wavelengths. The image of the ocular fundus was analyzed to find out if quantitative measurements of the reflectivity of tissue permit assessment of the oxygen perfusion of tissue. We explored problems that affect the reproducibility of patient measurements such as non-uniformity errors on the image. For the first time, an image processing technique was designed and used to minimize the errors of oxygen saturation measurements by illumination correction in retina wide field by increasing SNR. Retinal images were taken from healthy and diabetic retinopathy eyes using the cSLO with a confocal aperture of 100 μm. The ratio image (RI) of red/IR, as oxygen saturation (SO2) index, was calculated for normal eyes. The image correction technique improved the reproducibility of the measurements. Average RI intensity variation of healthy retina tissue was determined within a range of about 5.5%. The capability of the new technique to discriminate oxygenation levels of retinal artery and vein was successfully demonstrated and showed good promise in the diagnosis of the perfused retina.

  19. The binding of cellulase variants to dislocations: a semi-quantitative analysis based on CLSM (confocal laser scanning microscopy) images

    DEFF Research Database (Denmark)

    Hidayat, Budi J.; Weisskopf, Carmen; Felby, Claus

    2015-01-01

    or slip planes. Here we study whether cellulases bind to dislocations to a higher extent than to the surrounding cell wall. The binding of fluorescently labelled cellobiohydrolases and endoglucanases to filter paper fibers was investigated using confocal laser scanning microscopy and a ratiometric method...

  20. The simplicity of males: Dwarf males of four species of Osedax (Siboglinidae; Annelida) investigated by confocal laser scanning microscopy

    DEFF Research Database (Denmark)

    Worsaae, Katrine; Rouse, Greg W

    2010-01-01

    . Here, we present the first investigation of the entire muscle and nervous system in dwarf males of Osedax frankpressi, O. roseus, O. rubiplumus, and O. spiral analyzed by multistaining and confocal laser scanning microscopy. Sperm shape and spermiogenesis, the sperm duct and internal and external...

  1. Elastomeric photo-actuators and their investigation by confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Czaniková, Klaudia; Ilčíková, Markéta; Mičušík, Matej; Kasák, Peter; Mosnáček, Jaroslav; Omastová, Mária; Krupa, Igor; Pavlova, Ewa; Chorvát Jr, Dušan

    2013-01-01

    The photo-actuation behavior of nanocomposites based on ethylene–vinylacetate copolymer (EVA) and styrene–isoprene–styrene (SIS) block copolymer filled with well-dispersed and modified multiwalled carbon nanotubes (MWCNTs) is discussed in this paper. The nanocomposites were prepared by casting from solution. To improve the dispersion of the MWCNTs in EVA, the MWCNT surface was modified with a non-covalent surfactant, cholesteryl 1-pyrenecarboxylate (PyChol). To prepare SIS nanocomposites, the MWCNT surface was covalently modified with polystyrene chains. The good dispersion of the filler was confirmed by transmission electron microscopy (TEM). Special, custom-made punch/die molds were used to create a Braille element (BE)-like shape, which under shear forces induces a uniaxial orientation of the MWCNTs within the matrix. The uniaxial orientation of MWCNTs is an essential precondition to ensure the photo-actuating behavior of MWCNTs in polymeric matrices. The orientation of the MWCNTs within the matrices was examined by scanning electron microscopy (SEM). Nanocomposite BEs were illuminated from the bottom by a red light-emitting diode (LED), and the photo-actuation was investigated by confocal laser scanning microscopy (CLSM). When the BEs were exposed to light, a temporary increase in the height of the element was detected. This process was observed to be reversible: after switching off the light, the BEs returned to their original shape and height. (paper)

  2. Using confocal laser scanning microscopy to probe the milk fat globule membrane and associated proteins.

    Science.gov (United States)

    Gallier, Sophie; Gragson, Derek; Jiménez-Flores, Rafael; Everett, David

    2010-04-14

    The bovine milk fat globule membrane (MFGM) is an important, biologically relevant membrane due to its functional and health properties. Its composition has been thoroughly studied, but its structure, especially the lateral organization of its components, still remains unclear. We have used confocal laser scanning microscopy (CLSM) to investigate the surface structure of the MFGM in globules with different degrees of processing using two types of fluorescently labeled phospholipid probes and a protein dye. Using this technique, we have observed heterogeneities in the distribution of MFGM lipids and proteins relating to the processing and size of the globules. The effect of pretreating the milk (centrifugation, pasteurization-homogenization and churning) was studied by double-staining the surface of the milk fat globules, followed by observation using CLSM, and by determining the phospholipid profile of raw milk, raw cream, processed milk and buttermilk powder. Our findings agree with other techniques by showing that the composition of the MFGM changes with processing through the loss of phospholipids and the adsorption of caseins and whey proteins onto the surface.

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

    Science.gov (United States)

    Skytte, Jacob L; Ghita, Ovidiu; Whelan, Paul F; Andersen, Ulf; Møller, Flemming; Dahl, Anders B; Larsen, Rasmus

    2015-06-01

    The microstructure of protein networks in yogurts defines important physical properties of the yogurt and hereby partly its quality. Imaging this protein network using confocal scanning laser microscopy (CSLM) has shown good results, and CSLM has become a standard measuring technique for fermented dairy products. When studying such networks, hundreds of images can be obtained, and here image analysis methods are essential for using the images in statistical analysis. Previously, methods including gray level co-occurrence matrix analysis and fractal analysis have been used with success. However, a range of other image texture characterization methods exists. These methods describe an image by a frequency distribution of predefined image features (denoted textons). Our contribution is an investigation of the choice of image analysis methods by performing a comparative study of 7 major approaches to image texture description. Here, CSLM images from a yogurt fermentation study are investigated, where production factors including fat content, protein content, heat treatment, and incubation temperature are varied. The descriptors are evaluated through nearest neighbor classification, variance analysis, and cluster analysis. Our investigation suggests that the texton-based descriptors provide a fuller description of the images compared to gray-level co-occurrence matrix descriptors and fractal analysis, while still being as applicable and in some cases as easy to tune. © 2015 Institute of Food Technologists®

  4. Apoplastic pH in corn root gravitropism: a laser scanning confocal microscopy measurement

    International Nuclear Information System (INIS)

    Taylor, D.P.; Slattery, J.; Leopold, A.C.

    1996-01-01

    The ability to measure the pH of the apoplast in situ is of special interest as a test of the cell wall acidification theory. Optical sectioning of living seedlings of corn roots using the laser scanning confocal microscope (LSCM) permits us to make pH measurements in living tissue. The pH of the apoplast of corn roots was measured by this method after infiltration with CI-NERF, a pH-sensitive dye, along with Texas Red Dextran 3000, a pH-insensitive dye, as an internal standard. In the elongation zone of corn roots, the mean apoplastic pH was 4.9. Upon gravitropic stimulation, the pH on the convex side of actively bending roots was 4.5. The lowering of the apoplastic pH by 0.4 units appears to be sufficient to account for the increased growth on that side. This technique provides site-specific evidence for the acid growth theory of cell elongation. The LSCM permits measurements of the pH of living tissues, and has a sensitivity of approximately 0.2 pH units. (author)

  5. A statistical pixel intensity model for segmentation of confocal laser scanning microscopy images.

    Science.gov (United States)

    Calapez, Alexandre; Rosa, Agostinho

    2010-09-01

    Confocal laser scanning microscopy (CLSM) has been widely used in the life sciences for the characterization of cell processes because it allows the recording of the distribution of fluorescence-tagged macromolecules on a section of the living cell. It is in fact the cornerstone of many molecular transport and interaction quantification techniques where the identification of regions of interest through image segmentation is usually a required step. In many situations, because of the complexity of the recorded cellular structures or because of the amounts of data involved, image segmentation either is too difficult or inefficient to be done by hand and automated segmentation procedures have to be considered. Given the nature of CLSM images, statistical segmentation methodologies appear as natural candidates. In this work we propose a model to be used for statistical unsupervised CLSM image segmentation. The model is derived from the CLSM image formation mechanics and its performance is compared to the existing alternatives. Results show that it provides a much better description of the data on classes characterized by their mean intensity, making it suitable not only for segmentation methodologies with known number of classes but also for use with schemes aiming at the estimation of the number of classes through the application of cluster selection criteria.

  6. Application of gold nanoparticles as contrast agents in confocal laser scanning microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lemelle, A; Veksler, B; Piletsky, S A; Meglinski, I [Cranfield Health, Cranfield University, Cranfield, MK43 0AL (United Kingdom); Kozhevnikov, I S; Akchurin, G G, E-mail: a.lemelle.s06@cranfield.ac.uk [Physics Faculty, Saratov State University, Saratov 410012 (Russian Federation)

    2009-01-15

    Confocal laser scanning microscopy (CLSM) is a modern high-resolution optical technique providing detailed image of tissue structure with high (down to microns) spatial resolution. Aiming at a concurrent improvement of imaging depth and image quality the CLSM requires the use of contrast agents. Commonly employed fluorescent contrast agents, such as fluorescent dyes and proteins, suffer from toxicity, photo-bleaching and overlapping with the tissues autofluorescence. Gold nanoparticles are potentially highly attractive to be applied as a contrast agent since they are not subject to photo-bleaching and can target biochemical cells markers associated with the specific diseases. In current report we consider the applicability of gold nano-spheres as a contrast agent to enhance quality of CLSM images of skin tissues in vitro versus the application of optical clearing agent, such as glycerol. The enhancement of CLSM image contrast was observed with an application of gold nano-spheres diffused within the skin tissues. We show that optical clearing agents such as a glycerol provide better CLSM image contrast than gold nano-spheres.

  7. Application of gold nanoparticles as contrast agents in confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Lemelle, A; Veksler, B; Piletsky, S A; Meglinski, I; Kozhevnikov, I S; Akchurin, G G

    2009-01-01

    Confocal laser scanning microscopy (CLSM) is a modern high-resolution optical technique providing detailed image of tissue structure with high (down to microns) spatial resolution. Aiming at a concurrent improvement of imaging depth and image quality the CLSM requires the use of contrast agents. Commonly employed fluorescent contrast agents, such as fluorescent dyes and proteins, suffer from toxicity, photo-bleaching and overlapping with the tissues autofluorescence. Gold nanoparticles are potentially highly attractive to be applied as a contrast agent since they are not subject to photo-bleaching and can target biochemical cells markers associated with the specific diseases. In current report we consider the applicability of gold nano-spheres as a contrast agent to enhance quality of CLSM images of skin tissues in vitro versus the application of optical clearing agent, such as glycerol. The enhancement of CLSM image contrast was observed with an application of gold nano-spheres diffused within the skin tissues. We show that optical clearing agents such as a glycerol provide better CLSM image contrast than gold nano-spheres

  8. Application of gold nanoparticles as contrast agents in confocal laser scanning microscopy

    Science.gov (United States)

    Lemelle, A.; Veksler, B.; Kozhevnikov, I. S.; Akchurin, G. G.; Piletsky, S. A.; Meglinski, I.

    2009-01-01

    Confocal laser scanning microscopy (CLSM) is a modern high-resolution optical technique providing detailed image of tissue structure with high (down to microns) spatial resolution. Aiming at a concurrent improvement of imaging depth and image quality the CLSM requires the use of contrast agents. Commonly employed fluorescent contrast agents, such as fluorescent dyes and proteins, suffer from toxicity, photo-bleaching and overlapping with the tissues autofluorescence. Gold nanoparticles are potentially highly attractive to be applied as a contrast agent since they are not subject to photo-bleaching and can target biochemical cells markers associated with the specific diseases. In current report we consider the applicability of gold nano-spheres as a contrast agent to enhance quality of CLSM images of skin tissues in vitro versus the application of optical clearing agent, such as glycerol. The enhancement of CLSM image contrast was observed with an application of gold nano-spheres diffused within the skin tissues. We show that optical clearing agents such as a glycerol provide better CLSM image contrast than gold nano-spheres.

  9. Confocal laser-scanning microscopy of capillaries in normal and psoriatic skin

    Science.gov (United States)

    Archid, Rami; Patzelt, Alexa; Lange-Asschenfeldt, Bernhard; Ahmad, Sufian S.; Ulrich, Martina; Stockfleth, Eggert; Philipp, Sandra; Sterry, Wolfram; Lademann, Juergen

    2012-10-01

    An important and most likely active role in the pathogenesis of psoriasis has been attributed to changes in cutaneous blood vessels. The purpose of this study was to use confocal laser-scanning microscopy (CLSM) to investigate dermal capillaries in psoriatic and normal skin. The structures of the capillary loops in 5 healthy participants were compared with those in affected skin of 13 psoriasis patients. The diameters of the capillaries and papillae were measured for each group with CLSM. All investigated psoriasis patients showed elongated, widened, and tortuous microvessels in the papillary dermis, whereas all healthy controls showed a single capillary loop in each dermal papilla. The capillaries of the papillary loop and the dermal papilla were significantly enlarged in the psoriatic skin lesions (diameters 24.39±2.34 and 146.46±28.52 μm, respectively) in comparison to healthy skin (diameters 9.53±1.8 and 69.48±17.16 μm, respectively) (P<0.001). CLSM appears to represent a promising noninvasive technique for evaluating dermal capillaries in patients with psoriasis. The diameter of the vessels could be seen as a well-quantifiable indicator for the state of psoriatic skin. CLSM could be useful for therapeutic monitoring to delay possible recurrences.

  10. In vivo assessment of the structure of skin microcirculation by reflectance confocal-laser-scanning microscopy

    Science.gov (United States)

    Sugata, Keiichi; Osanai, Osamu; Kawada, Hiromitsu

    2012-02-01

    One of the major roles of the skin microcirculation is to supply oxygen and nutrition to the surrounding tissue. Regardless of the close relationship between the microcirculation and the surrounding tissue, there are few non-invasive methods that can evaluate both the microcirculation and its surrounding tissue at the same site. We visualized microcapillary plexus structures in human skin using in vivo reflectance confocal-laser-scanning microscopy (CLSM), Vivascope 3000® (Lucid Inc., USA) and Image J software (National Institutes of Health, USA) for video image processing. CLSM is a non-invasive technique that can visualize the internal structure of the skin at the cellular level. In addition to internal morphological information such as the extracellular matrix, our method reveals capillary structures up to the depth of the subpapillary plexus at the same site without the need for additional optical systems. Video images at specific depths of the inner forearm skin were recorded. By creating frame-to-frame difference images from the video images using off-line video image processing, we obtained images that emphasize the brightness depending on changes of intensity coming from the movement of blood cells. Merging images from different depths of the skin elucidates the 3-dimensional fine line-structure of the microcirculation. Overall our results show the feasibility of a non-invasive, high-resolution imaging technique to characterize the skin microcirculation and the surrounding tissue.

  11. Combining confocal laser scanning microscopy with serial section reconstruction in the study of adult neurogenesis.

    Directory of Open Access Journals (Sweden)

    Federico eLuzzati

    2011-05-01

    Full Text Available Current advances in imaging techniques have extended the possibility of visualizing small structures within large volumes of both fixed and live specimens without sectioning. These techniques have contributed valuable information to study neuronal plasticity in the adult brain. However, technical limits still hamper the use of these approaches to investigate neurogenic regions located far from the ventricular surface such as parenchymal neurogenic niches, or the scattered neuroblasts induced by brain lesions. Here, we present a method to combine confocal laser scanning microscopy (CLSM and serial section reconstruction in order to reconstruct large volumes of brain tissue at cellular resolution. In this method a series of thick sections are imaged with CLSM and the resulting stacks of images are registered and 3D reconstructed. This approach is based on existing freeware software and can be performed on ordinary laboratory personal computers (PC. By using this technique we have investigated the morphology and spatial organization of a group of doublecortin (DCX+ neuroblasts located in the lateral striatum of the late post-natal guinea pig. The 3D study unravelled a complex network of long and poorly ramified cell processes, often fascicled and mostly oriented along the internal capsule fibre bundles. These data support CLSM serial section reconstruction as a reliable alternative to the whole mount approaches to analyze cyto-architectural features of adult germinative niches.

  12. Correlative Analysis of Immunoreactivity in Confocal Laser-Scanning Microscopy and Scanning Electron Microscopy with Focused Ion Beam Milling

    Directory of Open Access Journals (Sweden)

    Takahiro eSonomura

    2013-02-01

    Full Text Available Three-dimensional reconstruction of ultrastructure of rat brain with minimal effort has recently been realized by scanning electron microscopy combined with focused ion beam milling (FIB-SEM. Because application of immunohistochemical staining to electron microscopy has a great advantage in that molecules of interest are specifically localized in ultrastructures, we here tried to apply immunocytochemistry to FIB-SEM and correlate immunoreactivity in confocal laser-scanning microcopy (CF-LSM with that in FIB-SEM. The dendrites of medium-sized spiny neurons in rat neostriatum were visualized with a recombinant viral vector, which labeled the infected neurons with membrane-targeted GFP in a Golgi stain-like fashion, and thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2. After detecting the sites of terminals apposed to the dendrites in CF-LSM, GFP and VGluT2 immunoreactivities were further developed for electron microscopy by the immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB methods, respectively. In the contrast-inverted FIB-SEM images, silver precipitation and DAB deposits were observed as fine dark grains and diffuse dense profiles, respectively, indicating that these immunoreactivities were easily recognizable as in the images of transmission electron microscopy. In the sites of interest, some appositions were revealed to display synaptic specialization of asymmetric type. The present method is thus useful in the three-dimensional analysis of immunocytochemically differentiated synaptic connection in the central neural circuit.

  13. Real time detection of antibody-antigen interaction using a laser scanning confocal imaging-surface plasmon resonance system

    International Nuclear Information System (INIS)

    Zhang Hong-Yan; Yang Li-Quan; Ning Ting-Yin; Liu Wei-Min; Sun Jia-Yu; Wang Peng-Fei; Meng Lan; Nie Jia-Cai

    2012-01-01

    A laser scanning confocal imaging-surface plasmon resonance (LSCI-SPR) instrument integrated with a wavelength-dependent surface plasmon resonance (SPR) sensor and a laser scanning confocal microscopy (LSCM) is built to detect the bonding process of human IgG and fluorescent-labeled affinity purified antibodies in real time. The shifts of resonant wavelength at different reaction time stages are obtained by SPR, corresponding well with the changes of the fluorescence intensity collected by using LSCM. The instrument shows the merits of the combination and complementation of the SPR and LSCM, with such advantages as quantificational analysis, high spatial resolution and real time monitor, which are of great importance for practical applications in biosensor and life science. (general)

  14. Central serous chorioretinopathy fundus autofluorescence comparison with two different confocal scanning laser ophthalmoscopes.

    Science.gov (United States)

    Nam, Ki Tae; Yun, Cheol Min; Kim, Jee Taek; Yang, Kyung-Sook; Kim, Hyun Joo; Kim, Seong-Woo; Oh, Jaeryung; Huh, Kuhl

    2015-12-01

    To compare the lesion characteristics of two different types of confocal scanning laser ophthalmoscopy (cSLO) autofluorescence (AF) images in central serous chorioretinopathy (CSC). The study included 63 eyes of 61 patients; 63 pairs of fundus autofluorescence (FAF) images were compared before CSC resolution in 63 eyes, FAF images of 31 eyes were also compared after CSC resolution. The lesion characteristics (brightness and composite pattern) were compared between Heidelberg Retina Angiograph 2 (HRA2; Heidelberg Engineering, Germany) and Optomap Tx (Optomap; Optos, Scotland) FAF images. The lesion composite pattern was categorized as diffuse or granular. Diffuse AF was defined as homogenously increased or decreased AF, and granular AF was defined as dot-like, coarse changes in AF. The mean disease duration and subretinal fluid (SRF) height in the spectral domain optical coherence tomography were compared according to the FAF image characteristics. Lesion brightness before CSC resolution was hypo-AF in 48 eyes (76.2 %), hyper-AF in three (4.8 %), and mixed-AF in 12 (19.0 %) in HRA2 FAF images. In comparison, nine (14.3 %) images were hypo-AF, 44 (69.8 %) were hyper-AF, and 10 (15.9 %) were mixed-AF in Optomap FAF images (P < 0.0001). There was no significant difference in lesion composite pattern between the two FAF image wavelengths. Patients with lesions that were hyper-AF in Optomap FAF and hypo-AF in HRA2 FAF had a shorter disease duration and greater SRF height (1 month, 281 um) than those who were hyper-AF in both Optomap and HRA2 images (26 months, 153 um; P = 0.004, 0.001). The two types of FAF images of CSC showed different lesion brightness before and after CSC resolution but demonstrated similar lesion composite patterns.

  15. Performance of confocal scanning laser tomograph Topographic Change Analysis (TCA) for assessing glaucomatous progression.

    Science.gov (United States)

    Bowd, Christopher; Balasubramanian, Madhusudhanan; Weinreb, Robert N; Vizzeri, Gianmarco; Alencar, Luciana M; O'Leary, Neil; Sample, Pamela A; Zangwill, Linda M

    2009-02-01

    To determine the sensitivity and specificity of confocal scanning laser ophthalmoscope's Topographic Change Analysis (TCA; Heidelberg Retina Tomograph [HRT]; Heidelberg Engineering, Heidelberg, Germany) parameters for discriminating between progressing glaucomatous and stable healthy eyes. The 0.90, 0.95, and 0.99 specificity cutoffs for various (n=70) TCA parameters were developed by using 1000 permuted topographic series derived from HRT images of 18 healthy eyes from Moorfields Eye Hospital, imaged at least four times. The cutoffs were then applied to topographic series from 36 eyes with known glaucomatous progression (by optic disc stereophotograph assessment and/or standard automated perimetry guided progression analysis, [GPA]) and 21 healthy eyes from the University of California, San Diego (UCSD) Diagnostic Innovations in Glaucoma Study (DIGS), all imaged at least four times, to determine TCA sensitivity and specificity. Cutoffs also were applied to 210 DIGS patients' eyes imaged at least four times with no evidence of progression (nonprogressed) by stereophotography or GPA. The TCA parameter providing the best sensitivity/specificity tradeoff using the 0.90, 0.95, and 0.99 cutoffs was the largest clustered superpixel area within the optic disc margin (CAREA(disc) mm(2)). Sensitivities/specificities for classifying progressing (by stereophotography and/or GPA) and healthy eyes were 0.778/0.809, 0.639/0.857, and 0.611/1.00, respectively. In nonprogressing eyes, specificities were 0.464, 0.570, and 0.647 (i.e., lower than in the healthy eyes). In addition, TCA parameter measurements of nonprogressing eyes were similar to those of progressing eyes. TCA parameters can discriminate between progressing and longitudinally observed healthy eyes. Low specificity in apparently nonprogressing patients' eyes suggests early progression detection using TCA.

  16. Thermal maturity of Tasmanites microfossils from confocal laser scanning fluorescence microscopy

    Science.gov (United States)

    Hackley, Paul C.; Kus, Jolanta

    2015-01-01

    We report here, for the first time, spectral properties of Tasmanites microfossils determined by confocal laser scanning fluorescence microscopy (CLSM, using Ar 458 nm excitation). The Tasmanites occur in a well-characterized natural maturation sequence (Ro 0.48–0.74%) of Devonian shale (n = 3 samples) from the Appalachian Basin. Spectral property λmax shows excellent agreement (r2 = 0.99) with extant spectra from interlaboratory studies which used conventional fluorescence microscopy techniques. This result suggests spectral measurements from CLSM can be used to infer thermal maturity of fluorescent organic materials in geologic samples. Spectra of regions with high fluorescence intensity at fold apices and flanks in individual Tasmanites are blue-shifted relative to less-deformed areas in the same body that have lower fluorescence intensity. This is interpreted to result from decreased quenching moiety concentration at these locations, and indicates caution is needed in the selection of measurement regions in conventional fluorescence microscopy, where it is common practice to select high intensity regions for improved signal intensity and better signal to noise ratios. This study also documents application of CLSM to microstructural characterization of Tasmanites microfossils. Finally, based on an extant empirical relation between conventional λmax values and bitumen reflectance, λmax values from CLSM of Tasmanites microfossils can be used to calculate a bitumen reflectance equivalent value. The results presented herein can be used as a basis to broaden the future application of CLSM in the geological sciences into hydrocarbon prospecting and basin analysis.

  17. Degeneration process of fungiform taste buds after severing the human chorda tympani nerve--observation by confocal laser scanning microscopy.

    Science.gov (United States)

    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.

  18. Simultaneous Confocal Scanning Laser Ophthalmoscopy Combined with High-Resolution Spectral-Domain Optical Coherence Tomography: A Review

    Directory of Open Access Journals (Sweden)

    Verônica Castro Lima

    2011-01-01

    Full Text Available We aimed to evaluate technical aspects and the clinical relevance of a simultaneous confocal scanning laser ophthalmoscope and a high-speed, high-resolution, spectral-domain optical coherence tomography (SDOCT device for retinal imaging. The principle of confocal scanning laser imaging provides a high resolution of retinal and choroidal vasculature with low light exposure. Enhanced contrast, details, and image sharpness are generated using confocality. The real-time SDOCT provides a new level of accuracy for assessment of the angiographic and morphological correlation. The combined system allows for simultaneous recordings of topographic and tomographic images with accurate correlation between them. Also it can provide simultaneous multimodal imaging of retinal pathologies, such as fluorescein and indocyanine green angiographies, infrared and blue reflectance (red-free images, fundus autofluorescence images, and OCT scans (Spectralis HRA + OCT; Heidelberg Engineering, Heidelberg, Germany. The combination of various macular diagnostic tools can lead to a better understanding and improved knowledge of macular diseases.

  19. Confocal scanning microscopy

    DEFF Research Database (Denmark)

    Bariani, Paolo

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

  20. The neuromuscular system of Pycnophyes kielensis (Kinorhyncha: Allomalorhagida) investigated by confocal laser scanning microscopy.

    Science.gov (United States)

    Altenburger, Andreas

    2016-01-01

    Kinorhynchs are ecdysozoan animals with a phylogenetic position close to priapulids and loriciferans. To understand the nature of segmentation within Kinorhyncha and to infer a probable ancestry of segmentation within the last common ancestor of Ecdysozoa, the musculature and the nervous system of the allomalorhagid kinorhynch Pycnophyes kielensis were investigated by use of immunohistochemistry, confocal laser scanning microscopy, and 3D reconstruction software. The kinorhynch body plan comprises 11 trunk segments. Trunk musculature consists of paired ventral and dorsal longitudinal muscles in segments 1-10 as well as dorsoventral muscles in segments 1-11. Dorsal and ventral longitudinal muscles insert on apodemes of the cuticle inside the animal within each segment. Strands of longitudinal musculature extend over segment borders in segments 1-6. In segments 7-10, the trunk musculature is confined to the segments. Musculature of the digestive system comprises a strong pharyngeal bulb with attached mouth cone muscles as well as pharyngeal bulb protractors and retractors. The musculature of the digestive system shows no sign of segmentation. Judged by the size of the pharyngeal bulb protractors and retractors, the pharyngeal bulb, as well as the introvert, is moved passively by internal pressure caused by concerted action of the dorsoventral muscles. The nervous system comprises a neuropil ring anterior to the pharyngeal bulb. Associated with the neuropil ring are flask-shaped serotonergic somata extending anteriorly and posteriorly. A ventral nerve cord is connected to the neuropil ring and runs toward the anterior until an attachment point in segment 1, and from there toward the posterior with one ganglion in segment 6. Segmentation within Kinorhyncha likely evolved from an unsegmented ancestor. This conclusion is supported by continuous trunk musculature in the anterior segments 1-6, continuous pharyngeal bulb protractors and retractors throughout the anterior

  1. The neuromuscular system of Pycnophyes kielensis (Kinorhyncha: Allomalorhagida investigated by confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Andreas Altenburger

    2016-11-01

    Full Text Available Abstract Background Kinorhynchs are ecdysozoan animals with a phylogenetic position close to priapulids and loriciferans. To understand the nature of segmentation within Kinorhyncha and to infer a probable ancestry of segmentation within the last common ancestor of Ecdysozoa, the musculature and the nervous system of the allomalorhagid kinorhynch Pycnophyes kielensis were investigated by use of immunohistochemistry, confocal laser scanning microscopy, and 3D reconstruction software. Results The kinorhynch body plan comprises 11 trunk segments. Trunk musculature consists of paired ventral and dorsal longitudinal muscles in segments 1–10 as well as dorsoventral muscles in segments 1–11. Dorsal and ventral longitudinal muscles insert on apodemes of the cuticle inside the animal within each segment. Strands of longitudinal musculature extend over segment borders in segments 1–6. In segments 7–10, the trunk musculature is confined to the segments. Musculature of the digestive system comprises a strong pharyngeal bulb with attached mouth cone muscles as well as pharyngeal bulb protractors and retractors. The musculature of the digestive system shows no sign of segmentation. Judged by the size of the pharyngeal bulb protractors and retractors, the pharyngeal bulb, as well as the introvert, is moved passively by internal pressure caused by concerted action of the dorsoventral muscles. The nervous system comprises a neuropil ring anterior to the pharyngeal bulb. Associated with the neuropil ring are flask-shaped serotonergic somata extending anteriorly and posteriorly. A ventral nerve cord is connected to the neuropil ring and runs toward the anterior until an attachment point in segment 1, and from there toward the posterior with one ganglion in segment 6. Conclusions Segmentation within Kinorhyncha likely evolved from an unsegmented ancestor. This conclusion is supported by continuous trunk musculature in the anterior segments 1–6, continuous

  2. Observation of regenerated fungiform taste buds after severing the chorda tympani nerve using confocal laser scanning microscopy in vivo.

    Science.gov (United States)

    Saito, Takehisa; Ito, Tetsufumi; Kato, Yuji; Yamada, Takechiyo; Manabe, Yasuhiro; Narita, Norihiko

    2014-03-01

    To evaluate whether regenerated fungiform taste buds after severing the chorda tympani nerve can be detected by confocal laser scanning microscopy in vivo. Retrospective study. University hospital. Six patients with a normal gustatory function (Group 1), 9 patients with taste function recovery after severing the CTN (Group 2), and 5 patients without taste function recovery (Group 3) were included. In Groups 2 and 3, canal wall up (closed) tympanoplasty or canal wall down with canal reconstruction tympanoplasty was performed in all patients. Diagnostic. The severed nerves were readapted or approximated on the temporalis muscle fascia used to reconstruct the eardrum during surgery. Preoperative and postoperative gustatory functions were assessed using electrogustometry. Twelve to 260 months after severing the CTN, the surface of the midlateral region of the tongue was observed with a confocal laser microscope. EGM thresholds showed no response 1 month after surgery in all patients of Groups 2 and 3. In Group 2, EGM thresholds showed recovery 1 to 2 years after surgery and before confocal microscopy (-1.3 ± 6.5 dB). There was a significant difference between Group 1 (-5.7 ± 2.0 dB; p taste buds were observed in each FP, and 55 (79.7%) of 69 FP contained at least 1 taste bud. The mean number of taste bud per papilla was 3.7 ± 3.6. In patients with a recovered taste function (Group 2), 0 to 8 taste buds were observed in each FP. In this group, 54 (56.2%) of 94 FP contained at least 1 taste bud. The mean number of taste bud per papilla was 2.0 ± 2.2 (p taste bud was observed. Regenerated fungiform taste bud could be observed in vivo using confocal laser scanning microscopy, indicating that regenerated taste bud can be detected without biopsy.

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

    Science.gov (United States)

    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)

  4. 3D imaging of cement-based materials at submicron resolution by combining laser scanning confocal microscopy with serial sectioning.

    Science.gov (United States)

    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. Penetration and binding of monoclonal antibody in human osteosarcoma multicell spheroids. Comparison of confocal laser scanning microscopy and autoadiography

    International Nuclear Information System (INIS)

    Hjelstuen, M.H.; Rasch-Halvorsen, K.; Brekken, C.; Bruland, Oe.; Davies, C. de L.

    1996-01-01

    Penetration and binding of monoclonal antibody (MAb) in multicell osteosarcoma spheroids have been studied by autoradiography and confocal laser scanning microscopy (CLSM). Optical sectioning of the 3-dimensional spheroids was performed by CLSM. Owing to attenuation of fluorescence intensity, FITC-labelled MAb could not be detected at depths greater than 60 μm within the spheroids. The antibody uptake seen in autoradiographs and CLSM images 60 μm within the spheroids were essentially identical. MAb had reached all parts of the spheroids within 6 h. Quantitative measurements of the fluorescence intensity of FITC-labelled MAb seen in confocal images and measurements of MAb bound per cell using flow cytometry, showed that maximum uptake was reached after 6 h. The possibility to perform both quantatitive and qualitative measurements makes CLSM a promising method for studying antibody uptake in thick tissue samples. (orig.)

  6. 3D Imaging of Porous Media Using Laser Scanning Confocal Microscopy with Application to Microscale Transport Processes

    Energy Technology Data Exchange (ETDEWEB)

    Fredrich, J.T.

    1999-02-10

    We present advances in the application of laser scanning confocal microscopy (LSCM) to image, reconstruct, and characterize statistically the microgeometry of porous geologic and engineering materials. We discuss technical and practical aspects of this imaging technique, including both its advantages and limitations. Confocal imaging can be used to optically section a material, with sub-micron resolution possible in the lateral and axial planes. The resultant volumetric image data, consisting of fluorescence intensities for typically {approximately}50 million voxels in XYZ space, can be used to reconstruct the three-dimensional structure of the two-phase medium. We present several examples of this application, including studying pore geometry in sandstone, characterizing brittle failure processes in low-porosity rock deformed under triaxial loading conditions in the laboratory, and analyzing the microstructure of porous ceramic insulations. We then describe approaches to extract statistical microgeometric descriptions from volumetric image data, and present results derived from confocal volumetric data sets. Finally, we develop the use of confocal image data to automatically generate a three-dimensional mesh for numerical pore-scale flow simulations.

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

    Science.gov (United States)

    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.

  8. Confocal laser endomicroscopy

    DEFF Research Database (Denmark)

    Karstensen, John Gásdal; Săftoiu, Adrian; Brynskov, Jørn

    2016-01-01

    BACKGROUND AND STUDY AIMS: Confocal laser endomicroscopy (CLE) has been shown to predict relapse in ulcerative colitis in remission, but little is currently known about its role in Crohn's disease. The aim of this study was to identify reproducible CLE features in patients with Crohn's disease...

  9. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: LASER POWER MEASUREMENTS

    Science.gov (United States)

    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 ...

  10. Spatially resolved analyses of uranium species using a coupled system made up of confocal laser-scanning microscopy (CLSM) and laser induced fluorescence spectroscopy (LIFS)

    International Nuclear Information System (INIS)

    Brockmann, S.; Grossmann, K.; Arnold, T.

    2014-01-01

    The fluorescent properties of uranium when excited by UV light are used increasingly for spectroscope analyses of uranium species within watery samples. Here, alongside the fluorescent properties of the hexavalent oxidation phases, the tetra and pentavalent oxidation phases also play an increasingly important role. The detection of fluorescent emission spectrums on solid and biological samples using (time-resolved) laser induced fluorescence spectroscopy (TRLFS or LIFS respectively) has, however, the disadvantage that no statements regarding the spatial localisation of the uranium can be made. However, particularly in complex, biological samples, such statements on the localisation of the uranium enrichment in the sample are desired, in order to e.g. be able to distinguish between intra and extra-cellular uranium bonds. The fluorescent properties of uranium (VI) compounds and minerals can also be used to detect their localisation within complex samples. So the application of fluorescent microscopic methods represents one possibility to localise and visualise uranium precipitates and enrichments in biological samples, such as biofilms or cells. The confocal laser-scanning microscopy (CLSM) is especially well suited to this purpose. Coupling confocal laser-scanning microscopy (CLSM) with laser induced fluorescence spectroscopy (LIFS) makes it possible to localise and visualise fluorescent signals spatially and three-dimensionally, while at the same time being able to detect spatially resolved, fluorescent-spectroscopic data. This technology is characterised by relatively low detection limits from up to 1.10 -6 M for uranium (VI) compounds within the confocal volume. (orig.)

  11. Cutting efficiency of apical preparation using ultrasonic tips with microprojections: confocal laser scanning microscopy study

    Directory of Open Access Journals (Sweden)

    Sang-Won Kwak

    2014-11-01

    Full Text Available Objectives The purpose of this study was to compare the cutting efficiency of a newly developed microprojection tip and a diamond-coated tip under two different engine powers. Materials and Methods The apical 3-mm of each root was resected, and root-end preparation was performed with upward and downward pressure using one of the ultrasonic tips, KIS-1D (Obtura Spartan or JT-5B (B&L Biotech Ltd.. The ultrasonic engine was set to power-1 or -4. Forty teeth were randomly divided into four groups: K1 (KIS-1D / Power-1, J1 (JT-5B / Power-1, K4 (KIS-1D / Power-4, and J4 (JT-5B / Power-4. The total time required for root-end preparation was recorded. All teeth were resected and the apical parts were evaluated for the number and length of cracks using a confocal scanning micrscope. The size of the root-end cavity and the width of the remaining dentin were recorded. The data were statistically analyzed using two-way analysis of variance and a Mann-Whitney test. Results There was no significant difference in the time required between the instrument groups, but the power-4 groups showed reduced preparation time for both instrument groups (p < 0.05. The K4 and J4 groups with a power-4 showed a significantly higher crack formation and a longer crack irrespective of the instruments. There was no significant difference in the remaining dentin thickness or any of the parameters after preparation. Conclusions Ultrasonic tips with microprojections would be an option to substitute for the conventional ultrasonic tips with a diamond coating with the same clinical efficiency.

  12. Cutting efficiency of apical preparation using ultrasonic tips with microprojections: confocal laser scanning microscopy study.

    Science.gov (United States)

    Kwak, Sang-Won; Moon, Young-Mi; Yoo, Yeon-Jee; Baek, Seung-Ho; Lee, WooCheol; Kim, Hyeon-Cheol

    2014-11-01

    The purpose of this study was to compare the cutting efficiency of a newly developed microprojection tip and a diamond-coated tip under two different engine powers. The apical 3-mm of each root was resected, and root-end preparation was performed with upward and downward pressure using one of the ultrasonic tips, KIS-1D (Obtura Spartan) or JT-5B (B&L Biotech Ltd.). The ultrasonic engine was set to power-1 or -4. Forty teeth were randomly divided into four groups: K1 (KIS-1D / Power-1), J1 (JT-5B / Power-1), K4 (KIS-1D / Power-4), and J4 (JT-5B / Power-4). The total time required for root-end preparation was recorded. All teeth were resected and the apical parts were evaluated for the number and length of cracks using a confocal scanning micrscope. The size of the root-end cavity and the width of the remaining dentin were recorded. The data were statistically analyzed using two-way analysis of variance and a Mann-Whitney test. There was no significant difference in the time required between the instrument groups, but the power-4 groups showed reduced preparation time for both instrument groups (p < 0.05). The K4 and J4 groups with a power-4 showed a significantly higher crack formation and a longer crack irrespective of the instruments. There was no significant difference in the remaining dentin thickness or any of the parameters after preparation. Ultrasonic tips with microprojections would be an option to substitute for the conventional ultrasonic tips with a diamond coating with the same clinical efficiency.

  13. Quantitative and Qualitative Aspects of Gas-Metal-Oxide Mass Transfer in High-Temperature Confocal Scanning Laser Microscopy

    Science.gov (United States)

    Piva, Stephano P. T.; Pistorius, P. Chris; Webler, Bryan A.

    2018-05-01

    During high-temperature confocal scanning laser microscopy (HT-CSLM) of liquid steel samples, thermal Marangoni flow and rapid mass transfer between the sample and its surroundings occur due to the relatively small sample size (diameter around 5 mm) and large temperature gradients. The resulting evaporation and steel-slag reactions tend to change the chemical composition in the metal. Such mass transfer effects can change observed nonmetallic inclusions. This work quantifies oxide-metal-gas mass transfer of solutes during HT-CSLM experiments using computational simulations and experimental data for (1) dissolution of MgO inclusions in the presence and absence of slag and (2) Ca, Mg-silicate inclusion changes upon exposure of a Si-Mn-killed steel to an oxidizing gas atmosphere.

  14. Noise analysis of a white-light supercontinuum light source for multiple wavelength confocal laser scanning fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    McConnell, Gail [Centre for Biophotonics, Strathclyde Institute for Biomedical Sciences, University of Strathclyde, 27 Taylor Street, Glasgow, G4 0NR (United Kingdom)

    2005-08-07

    Intensity correlations of a Ti : sapphire, Kr/Ar and a white-light supercontinuum were performed to quantify the typical signal amplitude fluctuations and hence ascertain the comparative output stability of the white-light supercontinuum source for confocal laser scanning microscopy (CLSM). Intensity correlations across a two-pixel sample (n = 1000) of up to 98%, 95% and 94% were measured for the Ti : sapphire, Kr/Ar and white-light supercontinuum source, respectively. The white-light supercontinuum noise level is therefore acceptable for CLSM, with the added advantage of wider wavelength flexibility over traditional CLSM excitation sources. The relatively low-noise white-light supercontinuum was then used to perform multiple wavelength sequential CLSM of guinea pig detrusor to confirm the reliability of the system and to demonstrate system flexibility.

  15. Superresolution size determination in fluorescence microscopy: A comparison between spatially modulated illumination and confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Spoeri, Udo; Failla, Antonio Virgilio; Cremer, Christoph

    2004-01-01

    Recently developed far field light optical methods are a powerful tool to analyze biological nanostructures and their dynamics, in particular including the interior of three-dimensionally conserved cells. In this article, the recently described method of spatially modulated illumination (SMI) microscopy has been further extended to the online determination of the extension of small, subwavelength sized, fluorescent objects (nanosizing). Using fluorescence excitation with 488 nm, the determination of fluorescent labeled object diameters down to 40 nm corresponding to about 1/12th of the wavelength used for one-photon excitation could be shown. The results of the SMI nanosizing procedure for a detailed, systematic variation of the object diameter are presented together with a fast algorithm for online size evaluation. In addition, we show a direct comparison of the diameter of 'colocalization volumes' between SMI nanosizing and conventional confocal laser scanning microscopy

  16. Molecular confocal laser endomicroscopy

    DEFF Research Database (Denmark)

    Karstensen, John Gásdal; Klausen, Pia Helene; Saftoiu, Adrian

    2014-01-01

    While flexible endoscopy is essential for macroscopic evaluation, confocal laser endomicroscopy (CLE) has recently emerged as an endoscopic method enabling visualization at a cellular level. Two systems are currently available, one based on miniprobes that can be inserted via a conventional...... during on-going endoscopy), a novel world of molecular evaluation opens up. The method of molecular CLE could potentially be used for estimating the expression of important receptors in carcinomas, subsequently resulting in immediate individualization of treatment regimens, but also for improving...

  17.   In situ identification of streptococci and other bacteria in initial dental biofilm by confocal laser scanning microscopy and fluorescence in situ hybridization

    DEFF Research Database (Denmark)

    Dige, Irene; Kilian, Mogens; Nilsson, Holger

    2007-01-01

    Confocal laser scanning microscopy (CLSM) has been employed as a method for studying intact natural biofilm. When combined with fluorescence in situ hybridization (FISH) it is possible to analyze spatial relationships and changes of specific members of microbial populations over time. The aim...

  18. Activity and three-dimensional distribution of toluene-degrading Pseudomonas putida in a multispecies biofilm assessed by quantitative in situ hybridization and scanning confocal laser microscopy

    DEFF Research Database (Denmark)

    Møller, Søren; Pedersen, Anne Rathmann; Poulsen, L.K.

    1996-01-01

    As a representative member of the toluene-degrading population in a biofilter for waste gas treatment, Pseudomonas putida was investigated with a 16S rRNA targeting probe, The three-dimensional distribution of P. putida was visualized in the biofilm matrix by scanning confocal laser microscopy...

  19. Evaluation of baseline structural factors for predicting glaucomatous visual-field progression using optical coherence tomography, scanning laser polarimetry and confocal scanning laser ophthalmoscopy.

    Science.gov (United States)

    Sehi, M; Bhardwaj, N; Chung, Y S; Greenfield, D S

    2012-12-01

    The objective of this study is to assess whether baseline optic nerve head (ONH) topography and retinal nerve fiber layer thickness (RNFLT) are predictive of glaucomatous visual-field progression in glaucoma suspect (GS) and glaucomatous eyes, and to calculate the level of risk associated with each of these parameters. Participants with ≥28 months of follow-up were recruited from the longitudinal Advanced Imaging for Glaucoma Study. All eyes underwent standard automated perimetry (SAP), confocal scanning laser ophthalmoscopy (CSLO), time-domain optical coherence tomography (TDOCT), and scanning laser polarimetry using enhanced corneal compensation (SLPECC) every 6 months. Visual-field progression was assessed using pointwise linear-regression analysis of SAP sensitivity values (progressor) and defined as significant sensitivity loss of >1 dB/year at ≥2 adjacent test locations in the same hemifield at P<0.01. Cox proportional hazard ratios (HR) were calculated to determine the predictive ability of baseline ONH and RNFL parameters for SAP progression using univariate and multivariate models. Seventy-three eyes of 73 patients (43 GS and 30 glaucoma, mean age 63.2±9.5 years) were enrolled (mean follow-up 51.5±11.3 months). Four of 43 GS (9.3%) and 6 of 30 (20%) glaucomatous eyes demonstrated progression. Mean time to progression was 50.8±11.4 months. Using multivariate models, abnormal CSLO temporal-inferior Moorfields classification (HR=3.76, 95% confidence interval (CI): 1.02-6.80, P=0.04), SLPECC inferior RNFLT (per -1 μm, HR=1.38, 95% CI: 1.02-2.2, P=0.02), and TDOCT inferior RNFLT (per -1 μm, HR=1.11, 95% CI: 1.04-1.2, P=0.001) had significant HRs for SAP progression. Abnormal baseline ONH topography and reduced inferior RNFL are predictive of SAP progression in GS and glaucomatous eyes.

  20. Demons registration for in vivo and deformable laser scanning confocal endomicroscopy

    Science.gov (United States)

    Chiew, Wei Ming; Lin, Feng; Seah, Hock Soon

    2017-09-01

    A critical effect found in noninvasive in vivo endomicroscopic imaging modalities is image distortions due to sporadic movement exhibited by living organisms. In three-dimensional confocal imaging, this effect results in a dataset that is tilted across deeper slices. Apart from that, the sequential flow of the imaging-processing pipeline restricts real-time adjustments due to the unavailability of information obtainable only from subsequent stages. To solve these problems, we propose an approach to render Demons-registered datasets as they are being captured, focusing on the coupling between registration and visualization. To improve the acquisition process, we also propose a real-time visual analytics tool, which complements the imaging pipeline and the Demons registration pipeline with useful visual indicators to provide real-time feedback for immediate adjustments. We highlight the problem of deformation within the visualization pipeline for object-ordered and image-ordered rendering. Visualizations of critical information including registration forces and partial renderings of the captured data are also presented in the analytics system. We demonstrate the advantages of the algorithmic design through experimental results with both synthetically deformed datasets and actual in vivo, time-lapse tissue datasets expressing natural deformations. Remarkably, this algorithm design is for embedded implementation in intelligent biomedical imaging instrumentation with customizable circuitry.

  1. In-situ observation of recrystallization in an AlMgScZr alloy using confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Taendl, J.; Nambu, S.; Orthacker, A.; Kothleitner, G.; Inoue, J.; Koseki, T.; Poletti, C.

    2015-01-01

    In this work we present a novel in-situ approach to study the recrystallization behavior of age hardening alloys. We use confocal laser scanning microscopy (CLSM) at 400 °C to investigate the static recrystallization of an AlMg4Sc0.4Zr0.12 alloy in-situ. The results are combined with electron backscatter diffraction (EBSD) and transmission electron microscopy (TEM) analyses. It was found that CLSM is a powerful tool to visualize both the local initiation and temporal sequence of recrystallization. After fast nucleation and initial growth, the grain growth rate decreases and the grain boundary migration stops after some minutes due to Zener pinning from Al 3 (Sc,Zr) precipitates produced during the heat treatment. EBSD and TEM analyses confirm both the boundary movements and the particle-boundary interactions. - Highlights: • First time that CLSM is used to study recrystallization in-situ. • The start and end of recrystallization can be directly observed. • The procedure is easy to apply and requires only simple data interpretation. • In-situ observations on the surface correlate to modifications inside the bulk. • In-situ observations correlate to EBSD and EFTEM analyses.

  2. Porosity of natural stone and use of confocal laser scanning microscopy on calcitic marble aged in laboratory

    Directory of Open Access Journals (Sweden)

    Ana Mladenovič

    2008-06-01

    Full Text Available Porosity is one of the key characteristics of natural stone, which influences ondurability as well as functionality of stone as building material. Further, deterioration processes themselves are also characterized by change of porosity. Different direct and indirect techniques can be used for porosity determination. In the following paper overview of these methods, as well as their advantages and disadvantages, is given. Confocal laser scanning microscopy (CLSM is indirect (microscopic technique. Despite its numerous advantages, among which 3D visualizationof pore structure is of major importance, this technique is less known in the area of building materials. An example how CLSM can be applied for qualitative and quantitative evaluation of porosity of calcitic polygonal granoblastic marble is given in this paper. Studied marble has been, despite of its poor durability, often used as building material, especially in the case of claddings. It is shown that thermal hydric factors of deterioration can influence porosity significantly,especially formation of intergranular cracks.This kind of deterioration can be successfully evaluated with use of CLSM method, if samples are suitable prepared and if suitable image analysis tools are developed.

  3. In-situ observation of recrystallization in an AlMgScZr alloy using confocal laser scanning microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Taendl, J., E-mail: johannes.taendl@tugraz.atl [Institute of Materials Science and Welding, Graz University of Technology, Graz (Austria); Nambu, S. [Department of Materials Engineering, The University of Tokyo, Tokyo 113-8656 (Japan); Orthacker, A.; Kothleitner, G. [Institute of Electron Microscopy and Nanoanalysis, Graz University of Technology, Graz (Austria); Graz Center for Electron Microscopy, Graz (Austria); Inoue, J.; Koseki, T. [Department of Materials Engineering, The University of Tokyo, Tokyo 113-8656 (Japan); Poletti, C. [Institute of Materials Science and Welding, Graz University of Technology, Graz (Austria)

    2015-10-15

    In this work we present a novel in-situ approach to study the recrystallization behavior of age hardening alloys. We use confocal laser scanning microscopy (CLSM) at 400 °C to investigate the static recrystallization of an AlMg4Sc0.4Zr0.12 alloy in-situ. The results are combined with electron backscatter diffraction (EBSD) and transmission electron microscopy (TEM) analyses. It was found that CLSM is a powerful tool to visualize both the local initiation and temporal sequence of recrystallization. After fast nucleation and initial growth, the grain growth rate decreases and the grain boundary migration stops after some minutes due to Zener pinning from Al{sub 3}(Sc,Zr) precipitates produced during the heat treatment. EBSD and TEM analyses confirm both the boundary movements and the particle-boundary interactions. - Highlights: • First time that CLSM is used to study recrystallization in-situ. • The start and end of recrystallization can be directly observed. • The procedure is easy to apply and requires only simple data interpretation. • In-situ observations on the surface correlate to modifications inside the bulk. • In-situ observations correlate to EBSD and EFTEM analyses.

  4. Quantifying migration and polarization of murine mesenchymal stem cells on different bone substitutes by confocal laser scanning microscopy.

    Science.gov (United States)

    Roldán, J C; Chang, E; Kelantan, M; Jazayeri, L; Deisinger, U; Detsch, R; Reichert, T E; Gurtner, G C

    2010-12-01

    Cell migration is preceded by cell polarization. The aim of the present study was to evaluate the impact of the geometry of different bone substitutes on cell morphology and chemical responses in vitro. Cell polarization and migration were monitored temporally by using confocal laser scanning microscopy (CLSM) to follow green fluorescent protein (GFP)±mesenchymal stem cells (MSCs) on anorganic cancellous bovine bone (Bio-Oss(®)), β-tricalcium phosphate (β-TCP) (chronOS(®)) and highly porous calcium phosphate ceramics (Friedrich-Baur-Research-Institute for Biomaterials, Germany). Differentiation GFP±MSCs was observed using pro-angiogenic and pro-osteogenic biomarkers. At the third day of culture polarized vs. non-polarized cellular sub-populations were clearly established. Biomaterials that showed more than 40% of polarized cells at the 3rd day of culture, subsequently showed an enhanced cell migration compared to biomaterials, where non-polarized cells predominated (ppolarization predominated at the 7th day of culture (p=0.001). This model opens an interesting approach to understand osteoconductivity at a cellular level. MSCs are promising in bone tissue engineering considering the strong angiogenic effect before differentiation occurs. Copyright © 2010 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

  5. High-Temperature Confocal Laser Scanning Microscopy Studies of Ferrite Formation in Inclusion-Engineered Steels: A Review

    Science.gov (United States)

    Mu, Wangzhong; Hedström, Peter; Shibata, Hiroyuki; Jönsson, Pär G.; Nakajima, Keiji

    2018-05-01

    The concepts of oxide metallurgy and inclusion engineering can be utilized to improve the properties of low-alloy steels. These concepts aim at controlling the formation of intragranular ferrite (IGF), often a desirable microstructure providing good mechanical properties without the need for expensive alloying elements. IGF formation is stimulated to occur at non-metallic inclusions and form an arrangement of fine, interlocking ferrite grains. A method that has contributed significantly to investigations in this field lately is high-temperature confocal laser scanning microscopy (HT-CLSM). HT-CLSM is suited for in situ studies of inclusion behavior in liquid steel and phase transformations in solid-state steel, where in particular, displacive phase transformations can be studied, since they provide sufficient topographic contrast. The purpose of the present report is to provide a brief review of the state of the art of HT-CLSM and its application for in situ observations of ferrite formation in inclusion-engineered steels. The scientific literature in this field is surveyed and supplemented by new work to reveal the capability of HT-CLSM as well as to discuss the effect of factors such as cooling rate and parent grain size on IGF formation and growth kinetics. The report concludes with an outlook on the opportunities and challenges of HT-CLSM for applications in oxide metallurgy.

  6. Precision automation of cell type classification and sub-cellular fluorescence quantification from laser scanning confocal images

    Directory of Open Access Journals (Sweden)

    Hardy Craig Hall

    2016-02-01

    Full Text Available While novel whole-plant phenotyping technologies have been successfully implemented into functional genomics and breeding programs, the potential of automated phenotyping with cellular resolution is largely unexploited. Laser scanning confocal microscopy has the potential to close this gap by providing spatially highly resolved images containing anatomic as well as chemical information on a subcellular basis. However, in the absence of automated methods, the assessment of the spatial patterns and abundance of fluorescent markers with subcellular resolution is still largely qualitative and time-consuming. Recent advances in image acquisition and analysis, coupled with improvements in microprocessor performance, have brought such automated methods within reach, so that information from thousands of cells per image for hundreds of images may be derived in an experimentally convenient time-frame. Here, we present a MATLAB-based analytical pipeline to 1 segment radial plant organs into individual cells, 2 classify cells into cell type categories based upon random forest classification, 3 divide each cell into sub-regions, and 4 quantify fluorescence intensity to a subcellular degree of precision for a separate fluorescence channel. In this research advance, we demonstrate the precision of this analytical process for the relatively complex tissues of Arabidopsis hypocotyls at various stages of development. High speed and robustness make our approach suitable for phenotyping of large collections of stem-like material and other tissue types.

  7. Mycelial pellet intrastructure visualization and viability prediction in a culture of Streptomyces fradiae using confocal scanning laser microscopy

    International Nuclear Information System (INIS)

    Park, Y.; Tamura, S.; Koike, Y.; Toriya, M.; Okabe, M.

    1997-01-01

    The intrastructure of mycelial pellets of Streptomyces fradiae, which produces tylosin, was visualized following labeling with fluorescein isothiocyanate (FITC) and propidium iodide (PI) using confocal scanning laser microscopy. A PI-labeled inactive core was present inside the mycelial pellet in both fermentor and air-lift reactor cultures. The thickness of the active mycelial layer on the pellet surface was calculated from a density sliced image to be 60 and 68 μm respectively, in the fermentor and in air-lift reactor cultures. Using image analysis, the active mycelial concentration of pellets in the fermentor culture was predicted to be 2.1 times higher than that in the air-lift reactor culture. The tylosin production rate in the fermentor reached 0.78 g/l/d, which was 2.5-fold that in the air-lift reactor culture. These results indicate that the higher tylosin production rate in the fermentor culture was due to the higher active mycelial concentration in the fermentor compared to that in the air-lift reactor. (author)

  8. Nano-zymography Using Laser-Scanning Confocal Microscopy Unmasks Proteolytic Activity of Cell-Derived Microparticles.

    Science.gov (United States)

    Briens, Aurélien; Gauberti, Maxime; Parcq, Jérôme; Montaner, Joan; Vivien, Denis; Martinez de Lizarrondo, Sara

    2016-01-01

    Cell-derived microparticles (MPs) are nano-sized vesicles released by activated cells in the extracellular milieu. They act as vectors of biological activity by carrying membrane-anchored and cytoplasmic constituents of the parental cells. Although detection and characterization of cell-derived MPs may be of high diagnostic and prognostic values in a number of human diseases, reliable measurement of their size, number and biological activity still remains challenging using currently available methods. In the present study, we developed a protocol to directly image and functionally characterize MPs using high-resolution laser-scanning confocal microscopy. Once trapped on annexin-V coated micro-wells, we developed several assays using fluorescent reporters to measure their size, detect membrane antigens and evaluate proteolytic activity (nano-zymography). In particular, we demonstrated the applicability and specificity of this method to detect antigens and proteolytic activities of tissue-type plasminogen activator (tPA), urokinase and plasmin at the surface of engineered MPs from transfected cell-lines. Furthermore, we were able to identify a subset of tPA-bearing fibrinolytic MPs using plasma samples from a cohort of ischemic stroke patients who received thrombolytic therapy and in an experimental model of thrombin-induced ischemic stroke in mice. Overall, this method is promising for functional characterization of cell-derived MPs.

  9. Electron microscopy of intermediate filaments: teaming up with atomic force and confocal laser scanning microscopy.

    Science.gov (United States)

    Kreplak, Laurent; Richter, Karsten; Aebi, Ueli; Herrmann, Harald

    2008-01-01

    Intermediate filaments (IFs) were originally discovered and defined by electron microscopy in myoblasts. In the following it was demonstrated and confirmed that they constitute, in addition to microtubules and microfilaments, a third independent, general filament system in the cytoplasm of most metazoan cells. In contrast to the other two systems, IFs are present in cells in two principally distinct cytoskeletal forms: (i) extended and free-running filament arrays in the cytoplasm that are integrated into the cytoskeleton by associated proteins of the plakin type; and (ii) a membrane- and chromatin-bound thin 'lamina' of a more or less regular network of interconnected filaments made from nuclear IF proteins, the lamins, which differ in several important structural aspects from cytoplasmic IF proteins. In man, more than 65 genes code for distinct IF proteins that are expressed during embryogenesis in various routes of differentiation in a tightly controlled manner. IF proteins exhibit rather limited sequence identity implying that the different types of IFs have distinct biochemical properties. Hence, to characterize the structural properties of the various IFs, in vitro assembly regimes have been developed in combination with different visualization methods such as transmission electron microscopy of fixed and negatively stained samples as well as methods that do not use staining such as scanning transmission electron microscopy (STEM) and cryoelectron microscopy as well as atomic force microscopy. Moreover, with the generation of both IF-type specific antibodies and chimeras of fluorescent proteins and IF proteins, it has become possible to investigate the subcellular organization of IFs by correlative fluorescence and electron microscopic methods. The combination of these powerful methods should help to further develop our understanding of nuclear architecture, in particular how nuclear subcompartments are organized and in which way lamins are involved.

  10. Potential of confocal laser scanning microscopy for non-invasive diagnostics of malignant epithelial skin tumors in the course of dermatoheliosis progression

    Directory of Open Access Journals (Sweden)

    E. S. Snarskaya

    2016-01-01

    Full Text Available Most cases of malignant epithelial skin neoplasms including actinic keratosis and basal cell carcinoma, which are characterized by the most complicated course and numerous clinical and morphological options, involve dermatoheliosis progression. The risk of actinic keratosis transformation into basal cell carcinoma varies from 0.1% to 20% and up to 80% in cases of multiple AK lesion foci. A non-invasive method known as reflectance confocal laser scanning microscopy is the most promising one for the purposes of early diagnostics of signs pointing at epithelial skin neoplasm development and makes it possible to monitor the tumor in progress in vivo to diagnose the presence of a pool of squamous cells on a timely basis. The confocal laser scanning microscopy method provides high-contrast images of for any horizontal-oriented morphologic structures in the epidermis and upper dermis with a resolution comparable to those characteristic of traditional optical microscopy of skin tissue samples. According to our data obtained as a result of studying dynamic changes and morphologic structures in actinic keratosis foci (50 cases using the confocal laser scanning microscopy method, we discovered a number of morphologic features, and their further analysis will distinguish the signs of progressing carcinogenesis in case of dermatoheliosis.

  11. In vitro studies of Rickettsia-host cell interactions: Confocal laser scanning microscopy of Rickettsia helvetica-infected eukaryotic cell lines.

    Science.gov (United States)

    Speck, Stephanie; Kern, Tanja; Aistleitner, Karin; Dilcher, Meik; Dobler, Gerhard; Essbauer, Sandra

    2018-02-01

    Rickettsia (R.) helvetica is the most prevalent rickettsia found in Ixodes ricinus ticks in Germany. Several studies reported antibodies against R. helvetica up to 12.5% in humans investigated, however, fulminant clinical cases are rare indicating a rather low pathogenicity compared to other rickettsiae. We investigated growth characteristics of R. helvetica isolate AS819 in two different eukaryotic cell lines with focus on ultra-structural changes of host cells during infection determined by confocal laser scanning microscopy. Further investigations included partially sequencing of rickA, sca4 and sca2 genes, which have been reported to encode proteins involved in cell-to-cell spread and virulence in some rickettsiae. R. helvetica grew constantly but slowly in both cell lines used. Confocal laser scanning microscopy revealed that the dissemination of R. helvetica AS819 in both cell lines was rather mediated by cell break-down and bacterial release than cell-to-cell spread. The cytoskeleton of both investigated eukaryotic cell lines was not altered. R. helvetica possesses rickA, but its expression is not sufficient to promote actin-based motility as demonstrated by confocal laser scanning microscopy. Hypothetical Sca2 and Sca4 proteins were deduced from nucleotide gene sequences but the predicted amino acid sequences were disrupted or truncated compared to other rickettsiae most likely resulting in non-functional proteins. Taken together, these results might give a first hint to the underlying causes of the reduced virulence and pathogenicity of R. helvetica.

  12. 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.

    Science.gov (United States)

    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.

  13. Effect of the menstrual cycle on the optic nerve head in diabetes: analysis by confocal scanning laser ophthalmoscopy.

    Science.gov (United States)

    Akar, Munire Erman; Yucel, Iclal; Erdem, Uzeyir; Taskin, Omur; Ozel, Alper; Akar, Yusuf

    2005-04-01

    The purpose of this study was to examine and compare menstrual-cycle-dependent topographic changes in the optic nerve head of normally menstruating women with different grades of type 2 diabetes mellitus. We studied the right eyes of 123 normally menstruating women (36 with severe nonproliferative diabetic retinopathy [NPDR], 42 with mild NPDR and 45 healthy subjects). All subjects underwent a complete ocular examination at baseline. At 4 hormonally distinct phases of the menstrual cycle (early follicular, late follicular, mid-luteal and late luteal), we analysed the topography of the optic nerve head, using a confocal scanning laser ophthalmoscope, and measured the serum levels of estradiol, progesterone and luteinizing hormone. We excluded from analysis the data for 8 patients with severe NPDR, 10 patients with mild NPDR and 15 control subjects who were lost to follow-up examinations during the menstrual cycle. The mean age and optic disc area did not differ significantly among the 3 groups. The duration of diabetes was significantly longer in the patients with severe NPDR than in those with mild NPDR (p cup-shape measure, linear cup/disc ratio, cup/disc area ratio and cup area in the late luteal phase compared with the other phases of the menstrual cycle (p menstrual cycle. Severe NPDR is associated with significant topographic changes in the rim and cup of the optic nerve head during the menstrual cycle. This must be considered in the evaluation of women with both diabetes and glaucoma. The normal fluctuations in serum sex hormone levels during the menstrual cycle of diabetic women seem to affect the optic nerve head more when the disease is advanced.

  14. Image-guided intraocular injection using multimodality optical coherence tomography and fluorescence confocal scanning laser ophthalmoscopy in rodent ophthalmological models

    Science.gov (United States)

    Terrones, Benjamin D.; Benavides, Oscar R.; Leeburg, Kelsey C.; Mehanathan, Sankarathi B.; Levine, Edward M.; Tao, Yuankai K.

    2018-02-01

    Intraocular injections are routinely performed for delivery of anti-VEGF and anti-inflammatory therapies in humans. While these injections are also performed in mice to develop novel models of ophthalmic diseases and screen novel therapeutics, the injection location and volume are not well-controlled and reproducible. We overcome limitations of conventional injections methods by developing a multimodality, long working distance, non-contact optical coherence tomography (OCT) and fluorescence confocal scanning laser ophthalmoscopy (cSLO) system for retinal imaging before and after injections. Our OCT+cSLO system combines a custom-built spectraldomain OCT engine (875+/-85 nm) with 125 kHz line-rate with a modified commercial cSLO with a maximum frame-rate of 30 fps (512 x 512 pix.). The system was designed for an overlapping OCT+cSLO field-of-view of 1.1 mm with a 7.76 mm working distance to the pupil. cSLO excitation light sources and filters were optimized for simultaneous GFP and tdTomato imaging. Lateral resolution was 3.02 µm for OCT and 2.74 μm for cSLO. Intravitreal injections of 5%, 10%, and 20% intralipid with Alex Fluor 488 were manually injected intraocularly in C57BL/6 mice. Post-injection imaging showed structural changes associated with retinal puncture, including the injection track, a retinal elevation, and detachment of the posterior hyaloid. OCT enables quantitative analysis of injection location and volumes whereas complementary cSLO improves specificity for identifying fluorescently labeled injected compounds and transgenic cells. The long working distance of our non-contact OCT+cSLO system is uniquely-suited for concurrent imaging with intraocular injections and may be applied for imaging of ophthalmic surgical dynamics and real-time image-guided injections.

  15. Effects of two desensitizing dentifrices on dentinal tubule occlusion with citric acid challenge: Confocal laser scanning microscopy study

    Directory of Open Access Journals (Sweden)

    Sneha Anil Rajguru

    2017-01-01

    Full Text Available Background: Dentin hypersensitivity results when patent tubules are exposed to pain-inducing external stimuli. Aim: This study aims to compare the effects of two desensitizing dentifrices containing NovaMin and arginine on dentinal tubule occlusion with and without citric acid challenge in vitro using confocal laser scanning microscopy (CLSM. Materials and Methods: Forty dentin discs were randomly divided into Groups I and II containing twenty specimens each, treated with NovaMin and arginine-containing dentifrices, respectively. Groups I and II were divided into subgroups A and B where IA and IIA underwent CLSM analysis to determine the percentage of tubule occlusion while IB and IIB underwent 0.3% citric acid challenge and CLSM analysis. A novel grading system was devised to categorize tubule occlusion. Results: In Group II, the percentage of occluded tubules was highest for IIA (72.25% ± 10.57% and least for IIB (42.55% ± 8.65% having statistical significance (P < 0.0005. In Group I, the difference between IA (49.9% ± 12.96% and IB (43.15% ± 12.43% was statistically insignificant (P = 0.249. On the comparison between IB and IIB statistically indifferent result was obtained (P = 0.901, whereas the difference between IA and IIA was statistically significant (P < 0.001. The results of grading system were for IA 50% of samples belonged to Grade 2, for IIA 60% - Grade 3, and for IB 70% and for IIB 90% - Grade 2. Conclusion: Dentinal tubule occlusion with arginine-containing dentifrice was significantly higher than NovaMin. However, it could not resist citric acid challenge as effectively as NovaMin. The effects of NovaMin were more sustainable as compared to arginine-containing dentifrice, thus proving to be a better desensitizing agent.

  16. Multimodality optical coherence tomography and fluorescence confocal scanning laser ophthalmoscopy in a zebrafish model of retinal vascular occlusion and remodeling

    Science.gov (United States)

    Li, Xiaoyue; Spitz, Kathleen; Bozic, Ivan; Tao, Yuankai K.

    2018-02-01

    Neovascularization in diabetic retinopathy (DR) and age-related macular degeneration (AMD) result in severe vision-loss and are two of the leading causes of blindness. The structural, metabolic, and vascular changes underlying retinal neovascularization are unknown and, thus, there is an unmet need to identify mechanisms of pathogenesis and novel anti-angiogenic therapies. Zebrafish is a robust ophthalmological model because its retina has comparable structure to the human retina and its fecundity and life-cycle enable development of mutant phenotypes of human pathologies. Here, we perform multimodal imaging with OCT and fluorescence confocal scanning laser ophthalmoscopy (cSLO) to identify changes in retinal structure and function in a zebrafish model of vascular leakage. Transgenic zebrafish with EGFP tagged plasma protein were imaged longitudinally at six time points over two weeks to visualize vascular perfusion changes from diethylaminobenzaldehyde (DEAB) treatment. Complementary contrast from OCT-A perfusion maps and cSLO imaging of plasma protein EGFP shows vascular occlusions posttreatment. cSLO images confirm presence of vessels despite loss of OCT-A signal. Plasma protein EGFP contrast also shows significant changes in vessel structure as compared to baseline images. OCT structural volumes show empty vessel cross-sections confirming non-perfusion. In addition, we present algorithms for automated biometric identification of OCT datasets using OCT-A vascular patterns in the presence of significant vascular perfusion changes. These results establish a framework for large-scale in vivo assays to identify novel anti-angiogenic compounds and understand the mechanisms ofneovascularization associated with retinal ocular pathologies.

  17. Tumor-specific antivascular effect of TZT-1027 (Soblidotin) elucidated by magnetic resonance imaging and confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Natsume, Tsugitaka; Watanabe, Junichi; Kobayashi, Motohiro; Ogawa, Kenji; Yasumura, Kazuhiko

    2007-01-01

    TZT-1027 (soblidotin), an antimicrotubule agent, has previously been evaluated in terms of its antivascular effects. In this study, Evans blue perfusion, magnetic resonance imaging (MRI), and confocal laser scanning microscopy (CLSM) were utilized to further elucidate the antivascular effect of TZT-1027 in female nude mice and rats bearing human breast tumor MX-1, as well as in female Sprague-Dawley rats that developed breast tumors induced by dimethylbenz(a)anthracene (DMBA). Therapeutic doses of TZT-1027 caused nearly complete regression of implanted MX-1 tumors in nude mice and rats as well as DMBA-induced tumors in rats. The perfusion in MX-1 tumor implanted in nude mice was drastically reduced within 30 min after TZT-1027 administration and was completely inhibited after 6 h or more, although not reduced in normal tissue of kidney. The study using MRI demonstrated that rich blood flow within tumors was remarkably reduced 1-3 h after TZT-1027 administration both in nude rats bearing MX-1 tumors and in rats with DMBA-induced tumors. Furthermore, the study with CLSM in nude mice bearing MX-1 tumors revealed a disruption of tumor microvessels at 1 h and a destruction of tumor microvessel network at 3 h after TZT-1027 administration. In contrast, these types of vascular disorders were not observed in heart and kidney. These results suggest that TZT-1027 specifically damages tumor vasculatures, leading to extensive tumor necrosis within tolerable dose range, and confirms earlier observations that TZT-1027 exerts a considerable antivascular effect in addition to an excellent cytotoxic effect. (author)

  18. Confocal scanning microscope for nuclear photoemulsion

    International Nuclear Information System (INIS)

    Batusov, Yu.A.; Kovalev, Yu.S.; Soroko, L.M.

    2005-01-01

    The application of the confocal scanning microscope to the objects in the nuclear photoemulsion is described. An array of 27 microtomograms of single silver grain is shown. The cross sections of the same particle track of diameter 1 μm, detected by means of the confocal scanning microscope with open and annular apertures, are presented. It was shown that the confocal scanning microscope opens indeed new opportunities for the nuclear photoemulsion technique to get previously inaccessible information for physics of the short-living particles

  19. Development of useful recombinant promoter and its expression analysis in different plant cells using confocal laser scanning microscopy.

    Directory of Open Access Journals (Sweden)

    Deepak Kumar

    Full Text Available BACKGROUND: Designing functionally efficient recombinant promoters having reduced sequence homology and enhanced promoter activity will be an important step toward successful stacking or pyramiding of genes in a plant cell for developing transgenic plants expressing desired traits(s. Also basic knowledge regarding plant cell specific expression of a transgene under control of a promoter is crucial to assess the promoter's efficacy. METHODOLOGY/PRINCIPAL FINDINGS: We have constructed a set of 10 recombinant promoters incorporating different up-stream activation sequences (UAS of Mirabilis mosaic virus sub-genomic transcript (MS8, -306 to +27 and TATA containing core domains of Figwort mosaic virus sub-genomic transcript promoter (FS3, -271 to +31. Efficacies of recombinant promoters coupled to GUS and GFP reporter genes were tested in tobacco protoplasts. Among these, a 369-bp long hybrid sub-genomic transcript promoter (MSgt-FSgt showed the highest activity in both transient and transgenic systems. In a transient system, MSgt-FSgt was 10.31, 2.86 and 2.18 times more active compared to the CaMV35S, MS8 and FS3 promoters, respectively. In transgenic tobacco (Nicotiana tabaccum, var. Samsun NN and Arabidopsis plants, the MSgt-FSgt hybrid promoter showed 14.22 and 7.16 times stronger activity compared to CaMV35S promoter respectively. The correlation between GUS activity and uidA-mRNA levels in transgenic tobacco plants were identified by qRT-PCR. Both CaMV35S and MSgt-FSgt promoters caused gene silencing but the degree of silencing are less in the case of the MSgt-FSgt promoter compared to CaMV35S. Quantification of GUS activity in individual plant cells driven by the MSgt-FSgt and the CaMV35S promoter were estimated using confocal laser scanning microscopy and compared. CONCLUSION AND SIGNIFICANCE: We propose strong recombinant promoter MSgt-FSgt, developed in this study, could be very useful for high-level constitutive expression of transgenes in

  20. Membrane Separated Flow Cell for Parallelized Electrochemical Impedance Spectroscopy and Confocal Laser Scanning Microscopy to Characterize Electro-Active Microorganisms

    International Nuclear Information System (INIS)

    Stöckl, Markus; Schlegel, Christin; Sydow, Anne; Holtmann, Dirk; Ulber, Roland; Mangold, Klaus-Michael

    2016-01-01

    Highlights: • Development of a membrane separated electrochemical flow cell. • Simultaneous combination of EIS and CLSM. • Monitoring of bacterial cell attachment to anode of MFC. • Cell attachment of Shewanella oneidensis is shown. - Abstract: Understanding the attachment of electro-active bacteria to electrode surfaces and their subsequent biofilm formation is one of the major challenges for the establishment of bacterial bioelectrochemial systems (BES). For a constant observation of biofilm growth, providing information on different stages of biofilm formation, continuous monitoring methods are required. In this paper a combination of two powerful analytical methods, Electrochemical Impedance Spectroscopy (EIS) and Confocal Laser Scanning Microscopy (CLSM), for biofilm monitoring is presented. A custom-built flow cell with a transparent indium tin oxide working electrode (WE) was constructed allowing monitoring of cell attachment to a working electrode simultaneously by EIS and CLSM. Cyclic Voltammetry (CV) and EIS of an iron (II)/iron (III) redox couple indicate that the flow cell is suitable for electrochemical experiments. An engineered Shewanella oneidensis MR-1 (ATCC700550) producing eGFP was used as electro-active model organism to demonstrate the practical application of the flow cell as BES to monitor cell attachment simultaneously with EIS and CLSM. Applying the flow cell as MFC (transparent working electrode poised as anode) produced a typical current curve for such a system. From the equivalent circuit used to interpret EIS data the charge transfer resistance R CT is sensitive to attachment of microorganisms. Fitted R CT was increased initially after cell inoculation and then lowered constantly with progressing experimental time. In parallel taken CLSM images show that bacteria already adhered to the WE 5 min after inoculation. A mono- respectively bilayer of electro-active cells was observed after 17 h on the WE surface. With the presented

  1. Distribution of biomolecules in porous nitrocellulose membrane pads using confocal laser scanning microscopy and high-speed cameras.

    Science.gov (United States)

    Mujawar, Liyakat Hamid; Maan, Abid Aslam; Khan, Muhammad Kashif Iqbal; Norde, Willem; van Amerongen, Aart

    2013-04-02

    The main focus of our research was to study the distribution of inkjet printed biomolecules in porous nitrocellulose membrane pads of different brands. We produced microarrays of fluorophore-labeled IgG and bovine serum albumin (BSA) on FAST, Unisart, and Oncyte-Avid slides and compared the spot morphology of the inkjet printed biomolecules. The distribution of these biomolecules within the spot embedded in the nitrocellulose membrane was analyzed by confocal laser scanning microscopy in the "Z" stack mode. By applying a "concentric ring" format, the distribution profile of the fluorescence intensity in each horizontal slice was measured and represented in a graphical color-coded way. Furthermore, a one-step diagnostic antibody assay was performed with a primary antibody, double-labeled amplicons, and fluorophore-labeled streptavidin in order to study the functionality and distribution of the immune complex in the nitrocellulose membrane slides. Under the conditions applied, the spot morphology and distribution of the primary labeled biomolecules was nonhomogenous and doughnut-like on the FAST and Unisart nitrocellulose slides, whereas a better spot morphology with more homogeneously distributed biomolecules was observed on the Oncyte-Avid slide. Similar morphologies and distribution patterns were observed when the diagnostic one-step nucleic acid microarray immunoassay was performed on these nitrocellulose slides. We also investigated possible reasons for the differences in the observed spot morphology by monitoring the dynamic behavior of a liquid droplet on and in these nitrocellulose slides. Using high speed cameras, we analyzed the wettability and fluid flow dynamics of a droplet on the various nitrocellulose substrates. The spreading of the liquid droplet was comparable for the FAST and Unisart slides but different, i.e., slower, for the Oncyte-Avid slide. The results of the spreading of the droplet and the penetration behavior of the liquid in the

  2. 3D digital image processing for biofilm quantification from confocal laser scanning microscopy: Multidimensional statistical analysis of biofilm modeling

    Science.gov (United States)

    Zielinski, Jerzy S.

    The dramatic increase in number and volume of digital images produced in medical diagnostics, and the escalating demand for rapid access to these relevant medical data, along with the need for interpretation and retrieval has become of paramount importance to a modern healthcare system. Therefore, there is an ever growing need for processed, interpreted and saved images of various types. Due to the high cost and unreliability of human-dependent image analysis, it is necessary to develop an automated method for feature extraction, using sophisticated mathematical algorithms and reasoning. This work is focused on digital image signal processing of biological and biomedical data in one- two- and three-dimensional space. Methods and algorithms presented in this work were used to acquire data from genomic sequences, breast cancer, and biofilm images. One-dimensional analysis was applied to DNA sequences which were presented as a non-stationary sequence and modeled by a time-dependent autoregressive moving average (TD-ARMA) model. Two-dimensional analyses used 2D-ARMA model and applied it to detect breast cancer from x-ray mammograms or ultrasound images. Three-dimensional detection and classification techniques were applied to biofilm images acquired using confocal laser scanning microscopy. Modern medical images are geometrically arranged arrays of data. The broadening scope of imaging as a way to organize our observations of the biophysical world has led to a dramatic increase in our ability to apply new processing techniques and to combine multiple channels of data into sophisticated and complex mathematical models of physiological function and dysfunction. With explosion of the amount of data produced in a field of biomedicine, it is crucial to be able to construct accurate mathematical models of the data at hand. Two main purposes of signal modeling are: data size conservation and parameter extraction. Specifically, in biomedical imaging we have four key problems

  3. In-vivo diagnosis and non-inasive monitoring of Imiquimod 5% cream for non-melanoma skin cancer using confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Dietterle, S; Lademann, J; Röwert-Huber, H-J; Stockfleth, E; Astner, S; Antoniou, C; Sterry, W

    2008-01-01

    Basal cell carcinoma (BCC) is the most common cutaneous malignancy with increasing incidence rates worldwide. A number of established treatments are available, including surgical excision. The emergence of new non-invasive treatment modalities has prompted the development of non-invasive optical devices for therapeutic monitoring and evaluating treatment efficacy. This study was aimed to evaluate the clinical applicability of a fluorescence confocal laser scanning microscope (CFLSM) for non-invasive therapeutic monitoring of basal cell carcinoma treated with Imiquimod (Aldara®) as topical immune-response modifier. Eight participants with a diagnosis of basal cell carcinoma (BCC) were enrolled in this investigation. Sequential evaluation during treatment with Imiquimod showed progressive normalization of the confocal histomorphologic parameters in correlation with normal skin. Confocal laser scanning microscopy was able to identify characteristic features of BCC and allowed the visualization of therapeutic effects over time. Thus our results indicate the clinical applicability of CFLSM imaging to evaluate treatment efficacy in vivo and non-invasively

  4. Re-scan confocal microscopy: scanning twice for better resolution.

    Science.gov (United States)

    De Luca, Giulia M R; Breedijk, Ronald M P; Brandt, Rick A J; Zeelenberg, Christiaan H C; de Jong, Babette E; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A; Stallinga, Sjoerd; Manders, Erik M M

    2013-01-01

    We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required.

  5. The fractional laser-induced coagulation zone characterized over time by laser scanning confocal microscopy-A proof of concept study.

    Science.gov (United States)

    Banzhaf, Christina A; Lin, Lynlee L; Dang, Nhung; Freeman, Michael; Haedersdal, Merete; Prow, Tarl W

    2018-01-01

    Ablative fractional laser (AFXL) is an acknowledged technique to increase uptake of topical agents in skin. Micro thermal ablation zones (MAZs) consist of ablated vertical channels surrounded by a coagulation zone (CZ). Laser scanning confocal microscopy (LSCM) images individual MAZs at 733 nm (reflectance confocal microscopy (RCM)). Further, LSCM can image sodium fluorescein (NaF) fluorescence with 488 nm excitation (fluorescence confocal microcopy (FCM)), a small hydrophilic test molecule (370 MW, log P -1.52), which may simulate uptake, bio-distribution and kinetics of small hydrophilic drugs. To explore LSCM for combined investigations of CZ thickness and uptake, bio-distribution and kinetics of NaF in AFXL-exposed skin. Excised human abdominal skin samples were exposed to AFXL (15 mJ/microbeam, 2% density) and NaF gel (1000 μg/ml, 10 μl/cm2) in six repetitions, including untreated control samples. CZ thickness and spatiotemporal fluorescence intensities (FI) were quantified up to four hours after NaF application by RCM and FCM. Test sites were scanned to a depth of 200 μm, quantifying thickness of skin compartments (stratum corneum, epidermis, upper dermis), individual CZ thicknesses and FI in CZ and surrounding skin. RCM images established skin morphology to a depth of 200 μm. The CZ thickness measurements were feasible to a depth of 50 μm, and remained unchanged over time at 50 μm (P > 0.5). FI were detected to a depth of 160 μm and remained constant in CZ up to four hours after NaF application (15 minutes: 79 AU (73-92 AU), 60 minutes: 72 AU (58-82 AU), four hours: 78 AU (71-90 AU), P > 0.1). In surrounding skin, FI increased significantly over time, but remained lower than FI in CZ (15 minutes: 21 AU (17-22 AU), 60 minutes: 21 AU (19-26 AU), four hours: 42 (31- 48 AU), P = 0.03). AFXL-processed skin generated higher FI compared to non-laser processed skin in epidermis and upper dermis at 60 minutes and four hours

  6. Chlorophyll and its degradation products in the two-spotted spider mite, Tetranychus urticae: observations using epifluorescence and confocal laser scanning microscopy.

    Science.gov (United States)

    Occhipinti, Andrea; Maffei, Massimo E

    2013-10-01

    Chlorophyll and chlorophyll degradation products were observed in the two-spotted spider mite (Tetranychus urticae) using epifluorescence microscopy (EFM) and confocal laser scanning microscopy (CLSM). A clear red fluorescence (EFM) and a fluorescence induced by a laser wavelength of 650 nm (CLSM) were observed. In the lateral caeca, in the ventriculus and in the excretory organ, a bright light blue fluorescence was observed in close association with chlorophyll by using EFM. The same material can be localized with CLSM by using a laser with a wavelength of 488 nm. By comparison with synthetic guanine, this bright fluorescence is supposed to be guanine. The presence of guanine fluorescence in the mite pellets confirms this hypothesis. A possible mechanism for guanine formation is discussed.

  7. Real-Time Demonstration of Split Skin Graft Inosculation and Integra Dermal Matrix Neovascularization Using Confocal Laser Scanning Microscopy

    Science.gov (United States)

    Greenwood, John; Amjadi, Mahyar; Dearman, Bronwyn; Mackie, Ian

    2009-01-01

    Objectives: During the first 48 hours after placement, an autograft “drinks” nutrients and dissolved oxygen from fluid exuding from the underlying recipient bed (“plasmatic imbibition”). The theory of inosculation (that skin grafts subsequently obtain nourishment via blood vessel “anastomosis” between new vessels invading from the wound bed and existing graft vessels) was hotly debated from the late 19th to mid-20th century. This study aimed to noninvasively observe blood flow in split skin grafts and Integra™ dermal regeneration matrix to provide further proof of inosculation and to contrast the structure of vascularization in both materials, reflecting mechanism. Methods: Observations were made both clinically and using confocal microscopy on normal skin, split skin graft, and Integra™. The VivaScope™ allows noninvasive, real-time, in vivo images of tissue to be obtained. Results: Observations of blood flow and tissue architecture in autologous skin graft and Integra™ suggest that 2 very different processes are occurring in the establishment of circulation in each case. Inosculation provides rapid circulatory return to skin grafts whereas slower neovascularization creates an unusual initial Integra™ circulation. Conclusions: The advent of confocal laser microscopy like the VivaScope 1500™, together with “virtual” journals such as ePlasty, enables us to provide exciting images and distribute them widely to a “reading” audience. The development of the early Integra™ vasculature by neovascularization results in a large-vessel, high-volume, rapid flow circulation contrasting markedly from the inosculatory process in skin grafts and the capillary circulation in normal skin and merits further (planned) investigation. PMID:19787028

  8. Relationship between gustatory function and average number of taste buds per fungiform papilla measured by confocal laser scanning microscopy in humans.

    Science.gov (United States)

    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.

  9. Scanning Color Laser Microscope

    Science.gov (United States)

    Awamura, D.; Ode, T.; Yonezawa, M.

    1988-01-01

    A confocal color laser microscope which utilizes a three color laser light source (Red: He-Ne, Green: Ar, Blue: Ar) has been developed and is finding useful applications in the semiconductor field. The color laser microscope, when compared to a conventional microscope, offers superior color separation, higher resolution, and sharper contrast. Recently some new functions including a Focus Scan Memory, a Surface Profile Measurement System, a Critical Dimension Measurement system (CD) and an Optical Beam Induced Current Function (OBIC) have been developed for the color laser microscope. This paper will discuss these new features.

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

    Science.gov (United States)

    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.

  11. Ultrasonically synthesized organic liquid-filled chitosan microcapsules: part 2: characterization using AFM (atomic force microscopy) and combined AFM-confocal laser scanning fluorescence microscopy.

    Science.gov (United States)

    Mettu, Srinivas; Ye, Qianyu; Zhou, Meifang; Dagastine, Raymond; Ashokkumar, Muthupandian

    2018-04-25

    Atomic Force Microscopy (AFM) is used to measure the stiffness and Young's modulus of individual microcapsules that have a chitosan cross-linked shell encapsulating tetradecane. The oil filled microcapsules were prepared using a one pot synthesis via ultrasonic emulsification of tetradecane and crosslinking of the chitosan shell in aqueous solutions of acetic acid. The concentration of acetic acid in aqueous solutions of chitosan was varied from 0.2% to 25% v/v. The effect of acetic acid concentration and size of the individual microcapsules on the strength was probed. The deformations and forces required to rupture the microcapsules were also measured. Three dimensional deformations of microcapsules under large applied loads were obtained by the combination of Laser Scanning Confocal Microscopy (LSCM) with Atomic Force Microscopy (AFM). The stiffness, and hence the modulus, of the microcapsules was found to decrease with an increase in size with the average stiffness ranging from 82 to 111 mN m-1 and average Young's modulus ranging from 0.4 to 6.5 MPa. The forces required to rupture the microcapsules varied from 150 to 250 nN with deformations of the microcapsules up to 62 to 110% relative to their radius, respectively. Three dimensional images obtained using laser scanning confocal microscopy showed that the microcapsules retained their structure and shape after being subjected to large deformations and subsequent removal of the loads. Based on the above observations, the oil filled chitosan crosslinked microcapsules are an ideal choice for use in the food and pharmaceutical industries as they would be able to withstand the process conditions encountered.

  12. The effect of compression on clinical diagnosis of glaucoma based on non-analyzed confocal scanning laser ophthalmoscopy images

    NARCIS (Netherlands)

    Abramoff, M.D.

    2006-01-01

    Knowledge of the effect of compression of ophthalmic images on diagnostic reading is essential for effective tele-ophthalmology applications. It was therefore with great anticipation that I read the article “The Effect of Compression on Clinical Diagnosis of Glaucoma Based on Non-analyzed Confocal

  13. Spatially resolved analyses of uranium species using a coupled system made up of confocal laser-scanning microscopy (CLSM) and laser induced fluorescence spectroscopy (LIFS); Ortsaufgeloeste Analyse von Uranspezies mittels einem Gekoppelten System aus Konfokaler Laser-Scanning Mikroskopie (CLSM) und Laser Induzierter Fluoreszenzspektroskopie (LIFS)

    Energy Technology Data Exchange (ETDEWEB)

    Brockmann, S. [Verein fuer Kernverfahrenstechnik und Analytik Rossendorf e.V. (VKTA), Dresden (Germany); Grossmann, K.; Arnold, T. [Helmholtz-Zentrum Dresden-Rossendorf e.V. (Germany). Inst. fuer Ressourcenoekologie

    2014-01-15

    The fluorescent properties of uranium when excited by UV light are used increasingly for spectroscope analyses of uranium species within watery samples. Here, alongside the fluorescent properties of the hexavalent oxidation phases, the tetra and pentavalent oxidation phases also play an increasingly important role. The detection of fluorescent emission spectrums on solid and biological samples using (time-resolved) laser induced fluorescence spectroscopy (TRLFS or LIFS respectively) has, however, the disadvantage that no statements regarding the spatial localisation of the uranium can be made. However, particularly in complex, biological samples, such statements on the localisation of the uranium enrichment in the sample are desired, in order to e.g. be able to distinguish between intra and extra-cellular uranium bonds. The fluorescent properties of uranium (VI) compounds and minerals can also be used to detect their localisation within complex samples. So the application of fluorescent microscopic methods represents one possibility to localise and visualise uranium precipitates and enrichments in biological samples, such as biofilms or cells. The confocal laser-scanning microscopy (CLSM) is especially well suited to this purpose. Coupling confocal laser-scanning microscopy (CLSM) with laser induced fluorescence spectroscopy (LIFS) makes it possible to localise and visualise fluorescent signals spatially and three-dimensionally, while at the same time being able to detect spatially resolved, fluorescent-spectroscopic data. This technology is characterised by relatively low detection limits from up to 1.10{sup -6} M for uranium (VI) compounds within the confocal volume. (orig.)

  14. Quantitative characterization of cleavage and hydrogen-assisted quasi-cleavage fracture surfaces with the use of confocal laser scanning microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Merson, E. [Institute of Advanced Technologies, Togliatti State University, 445667 (Russian Federation); Kudrya, A.V.; Trachenko, V.A. [Department of Physical Metallurgy and the Physics of Strength, NUST MISiS, Moscow 119490 (Russian Federation); Merson, D. [Institute of Advanced Technologies, Togliatti State University, 445667 (Russian Federation); Laboratory for Advanced Materials, Kazan Federal University, Naberezhnye Chelny 423812, Republic of Tatarstan (Russian Federation); Danilov, V. [Institute of Advanced Technologies, Togliatti State University, 445667 (Russian Federation); Vinogradov, A. [Institute of Advanced Technologies, Togliatti State University, 445667 (Russian Federation); Department of Engineering Design and Materials, Norwegian University of Science and Technology – NTNU, N-7491 Trondheim (Norway)

    2016-05-17

    “True” cleavage (TC) and quasi-cleavage (QC) fracture surfaces of low-carbon steel specimens tested in liquid nitrogen and after hydrogen charging respectively were investigated by quantitative confocal laser scanning microscopy (CLSM) and conventional scanning electron microscopy (SEM) with electron-backscattered diffraction (EBSD). Topological and crystallographic features of the TC fracture surface are found in good agreement with the generally accepted cleavage mechanism: TC facets diameters correspond to those of grains; the crack path strictly follows the crystallographic orientation of grains and the most of the cleavage cracks are parallel to {100} planes. On the 2D SEM images, the QC facets appeared resembling the TC ones in terms of river line patterns, shapes and sizes. However, the substantial differences between the topography of these two kinds of fracture surfaces were revealed by 3D CLSM: the average misorientation angle between QC facets and the roughness of the QC fracture surface were much lower than those measured for TC. It is demonstrated that all these features are attributed to the specific fracture mechanism operating during hydrogen-assisted cracking.

  15. Quantitative characterization of cleavage and hydrogen-assisted quasi-cleavage fracture surfaces with the use of confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Merson, E.; Kudrya, A.V.; Trachenko, V.A.; Merson, D.; Danilov, V.; Vinogradov, A.

    2016-01-01

    “True” cleavage (TC) and quasi-cleavage (QC) fracture surfaces of low-carbon steel specimens tested in liquid nitrogen and after hydrogen charging respectively were investigated by quantitative confocal laser scanning microscopy (CLSM) and conventional scanning electron microscopy (SEM) with electron-backscattered diffraction (EBSD). Topological and crystallographic features of the TC fracture surface are found in good agreement with the generally accepted cleavage mechanism: TC facets diameters correspond to those of grains; the crack path strictly follows the crystallographic orientation of grains and the most of the cleavage cracks are parallel to {100} planes. On the 2D SEM images, the QC facets appeared resembling the TC ones in terms of river line patterns, shapes and sizes. However, the substantial differences between the topography of these two kinds of fracture surfaces were revealed by 3D CLSM: the average misorientation angle between QC facets and the roughness of the QC fracture surface were much lower than those measured for TC. It is demonstrated that all these features are attributed to the specific fracture mechanism operating during hydrogen-assisted cracking.

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

    Science.gov (United States)

    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 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.

  17. Attempt of correlative observation of morphological synaptic connectivity by combining confocal laser-scanning microscope and FIB-SEM for immunohistochemical staining technique.

    Science.gov (United States)

    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.

  18. Multimodality optical coherence tomography and fluorescence confocal scanning laser ophthalmoscopy for image-guided feedback of intraocular injections in mouse models

    Science.gov (United States)

    Benavides, Oscar R.; Terrones, Benjamin D.; Leeburg, Kelsey C.; Mehanathan, Sankarathi B.; Levine, Edward M.; Tao, Yuankai K.

    2018-02-01

    Rodent models are robust tools for understanding human retinal disease and function because of their similarities with human physiology and anatomy and availability of genetic mutants. Optical coherence tomography (OCT) has been well-established for ophthalmic imaging in rodents and enables depth-resolved visualization of structures and image-based surrogate biomarkers of disease. Similarly, fluorescence confocal scanning laser ophthalmoscopy (cSLO) has demonstrated utility for imaging endogenous and exogenous fluorescence and scattering contrast in the mouse retina. Complementary volumetric scattering and en face fluorescence contrast from OCT and cSLO, respectively, enables cellular-resolution longitudinal imaging of changes in ophthalmic structure and function. We present a non-contact multimodal OCT+cSLO small animal imaging system with extended working distance to the pupil, which enables imaging during and after intraocular injection. While injections are routinely performed in mice to develop novel models of ophthalmic diseases and screen novel therapeutics, the location and volume delivered is not precisely controlled and difficult to reproduce. Animals were imaged using a custom-built OCT engine and scan-head combined with a modified commercial cSLO scan-head. Post-injection imaging showed structural changes associated with retinal puncture, including the injection track, a retinal elevation, and detachment of the posterior hyaloid. When combined with imagesegmentation, we believe OCT can be used to precisely identify injection locations and quantify injection volumes. Fluorescence cSLO can provide complementary contrast for either fluorescently labeled compounds or transgenic cells for improved specificity. Our non-contact OCT+cSLO system is uniquely-suited for concurrent imaging with intraocular injections, which may be used for real-time image-guided injections.

  19. The effect of milk processing on the microstructure of the milk fat globule and rennet induced gel observed using confocal laser scanning microscopy.

    Science.gov (United States)

    Ong, L; Dagastine, R R; Kentish, S E; Gras, S L

    2010-04-01

    Confocal laser scanning microscopy (CLSM) was successfully used to observe the effect of milk processing on the size and the morphology of the milk fat globule in raw milk, raw ultrafiltered milk, and standardized and pasteurized milk prepared for cheese manufacture (cheese-milk) and commercial pasteurized and homogenized milk. Fat globule size distributions for the milk preparations were analyzed using both image analysis and light scattering and both measurements produced similar data trends. Changes to the native milk fat globule membrane (MFGM) were tracked using a MFGM specific fluorescent stain that allowed MFGM proteins and adsorbed proteins to be differentiated on the fat globule surface. Sodium dodecyl sulfate polyacrylamide gel electrophoresis confirmed the identity of native MFGM proteins isolated from the surface of fat globules within raw, UF retentate, and cheese-milk preparations, whereas only casein was detected on the surface of fat globules in homogenized milk. The microstructure, porosity, and gel strength of the rennet induced gel made from raw milk and cheese-milk was also found to be comparable and significantly different to that made from homogenized milk. Our results highlight the potential use of CLSM as a tool to observe the structural details of the fat globule and associated membrane close to its native environment.

  20. Ultrasensitive and selective gold film-based detection of mercury (II) in tap water using a laser scanning confocal imaging-surface plasmon resonance system in real time.

    Science.gov (United States)

    Zhang, Hongyan; Yang, Liquan; Zhou, Bingjiang; Liu, Weimin; Ge, Jiechao; Wu, Jiasheng; Wang, Ying; Wang, Pengfei

    2013-09-15

    An ultrasensitive and selective detection of mercury (II) was investigated using a laser scanning confocal imaging-surface plasmon resonance system (LSCI-SPR). The detection limit was as low as 0.01ng/ml for Hg(2+) ions in ultrapure and tap water based on a T-rich, single-stranded DNA (ssDNA)-modified gold film, which can be individually manipulated using specific T-Hg(2+)-T complex formation. The quenching intensity of the fluorescence images for rhodamine-labeled ssDNA fitted well with the changes in SPR. The changes varied with the Hg(2+) ion concentration, which is unaffected by the presence of other metal ions. The coefficients obtained for ultrapure and tap water were 0.99902 and 0.99512, respectively, for the linear part over a range of 0.01-100ng/ml. The results show that the double-effect sensor has potential for practical applications with ultra sensitivity and selectivity, especially in online or real-time monitoring of Hg(2+) ions pollution in tap water with the further improvement of portable LSCI-SPR instrument. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Evaluation of transdermal delivery of nanoemulsions in ex vivo porcine skin using two-photon microscopy and confocal laser-scanning microscopy

    Science.gov (United States)

    Choi, Sanghoon; Kim, Jin Woong; Lee, Yong Joong; Delmas, Thomas; Kim, Changhwan; Park, Soyeun; Lee, Ho

    2014-10-01

    This study experimentally evaluates the self-targeting ability of asiaticoside-loaded nanoemulsions compared with nontargeted nanoemulsions in ex vivo experiments with porcine skin samples. Homebuilt two-photon and confocal laser-scanning microscopes were employed to noninvasively examine the transdermal delivery of two distinct nanoemulsions. Prior to the application of nanoemulsions, we noninvasively observed the morphology of porcine skin using two-photon microscopy. We have successfully visualized the distributions of the targeted and nontargeted nanoemulsions absorbed into the porcine skin samples. Asiaticoside-loaded nanoemulsions showed an improved ex vivo transdermal delivery through the stratum corneum compared with nonloaded nanoemulsions. As a secondary measure, nanoemulsions-applied samples were sliced in the depth direction with a surgical knife in order to obtain the complete depth-direction distribution profile of Nile red fluorescence. XZ images demonstrated that asiaticoside-loaded nanoemulsion penetrated deeper into the skin compared with nontargeted nanoemulsions. The basal layer boundary is clearly visible in the case of the asiaticoside-loaded skin sample. These results reaffirm the feasibility of using self-targeting ligands to improve permeation through the skin barrier for cosmetics and topical drug applications.

  2. Comparison of fungiform taste-bud distribution among age groups using confocal laser scanning microscopy in vivo in combination with gustatory function.

    Science.gov (United States)

    Saito, Takehisa; Ito, Tetsufumi; Ito, Yumi; Manabe, Yasuhiro; Sano, Kazuo

    2016-04-01

    The aim of this study was to compare the distribution of taste buds in fungiform papillae (FP) and gustatory function between young and elderly age groups. Confocal laser scanning microscopy was used because it allows many FP to be observed non-invasively in a short period of time. The age of participants (n = 211) varied from 20 to 83 yr. The tip and midlateral region of the tongue were observed. Taste buds in an average of 10 FP in each area were counted. A total of 2,350 FP at the tongue tip and 2,592 FP in the midlateral region could be observed. The average number of taste buds was similar among all age groups both at the tongue tip and in the midlateral region. The taste function, measured by electrogustometry, among participants 20-29 yr of age was significantly lower than that in the other age groups; however, there was no difference among any other age groups in taste function. These results indicate that the peripheral gustatory system is well maintained anatomically and functionally in elderly people. © 2016 Eur J Oral Sci.

  3. Reproductive system abnormalities in Schistosoma mansoni adult worms isolated from Nectomys squamipes (Muridae: Sigmodontinae: brightfield and confocal laser scanning microscopy analysis

    Directory of Open Access Journals (Sweden)

    Neves Renata Heisler

    2003-01-01

    Full Text Available Schistosoma mansoni adult worms with genital anomalies isolated from Nectomys squamipes (Muridae: Sigmodontinae were studied by confocal laser scanning microscopy under the reflected mode. One male without testicular lobes (testicular agenesia/anorchism and two females, one with an atrophied ovary and another with 17 uterine eggs, were identified. The absence of testicular lobes occurred in a worm presenting otherwise normal male adult characteristics: tegument, tubercles and a gynaecophoric canal with spines. In both female specimens the digestive tube showed a vacuolated appearance, and the specimen with supernumerary uterine eggs exhibited a developing miracidium and an egg with a formed shell. The area of the ventral sucker was similar in both specimens however the tegument thickness, ovary and vitelline glands of the specimen with the atrophied ovary were smaller than those of the one with supernumerary eggs. These reported anomalies in the reproductive system call attention to the need to improve our understanding of genetic regulation and the possible role of environmental influences upon trematode development.

  4. Morphological aspects of Schistosoma mansoni adult worms isolated from nourished and undernourished mice: a comparative analysis by confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Neves Renata Heisler

    2001-01-01

    Full Text Available Malnutrition hampers the course of schistosomiasis mansoni infection just as normal growth of adult worms. A comparative morphometric study on adult specimens (male and female recovered from undernourished (fed with a low protein diet - regional basic diet and nourished (rodent commercial laboratory food, NUVILAB white mice was performed. Tomographic images and morphometric analysis of the oral and ventral suckers, reproductive system and tegument were obtained by means of confocal laser scanning microscopy. Undernourished male specimens presented smaller morphometric values (length and width of the reproductive system (first, third and last testicular lobes and thickness of the tegument than controls. Besides that, it was demonstrated that the dorsal surface of the male worms bears large tubercles unevenly distributed, but kept grouped and flat. At the subtegumental region, vacuolated areas were detected. It was concluded that the inadequate nutritional status of the vertebrate host has a negative influence mainly in the reproductive system and topographical somatic development of male adult Schistosoma mansoni, inducing some alterations on the structure of the parasite.

  5. A new improved protocol for in vitro intratubular dentinal bacterial contamination for antimicrobial endodontic tests: standardization and validation by confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Flaviana Bombarda de ANDRADE

    2015-01-01

    Full Text Available Objectives To compare three methods of intratubular contamination that simulate endodontic infections using confocal laser scanning microscopy (CLSM. Material and Methods Two pre-existing models of dentinal contamination were used to induce intratubular infection (groups A and B. These methods were modified in an attempt to improve the model (group C. Among the modifications it may be included: specimen contamination for five days, ultrasonic bath with BHI broth after specimen sterilization, use of E. faecalis during the exponential growth phase, greater concentration of inoculum, and two cycles of centrifugation on alternate days with changes of culture media. All specimens were longitudinally sectioned and stained with of LIVE/DEAD® for 20 min. Specimens were assessed using CLSM, which provided images of the depth of viable bacterial proliferation inside the dentinal tubules. Additionally, three examiners used scores to classify the CLSM images according to the following parameters: homogeneity, density, and depth of the bacterial contamination inside the dentinal tubules. Kruskal-Wallis and Dunn’s tests were used to evaluate the live and dead cells rates, and the scores obtained. Results The contamination scores revealed higher contamination levels in group C when compared with groups A and B (p0.05. The volume of live cells in group C was higher than in groups A and B (p<0.05. Conclusion The new protocol for intratubular infection resulted in high and uniform patterns of bacterial contamination and higher cell viability in all specimens when compared with the current methods.

  6. Assessment of statistical agreement of three techniques for the study of cut marks: 3D digital microscope, laser scanning confocal microscopy and micro-photogrammetry.

    Science.gov (United States)

    Maté-González, Miguel Ángel; Aramendi, Julia; Yravedra, José; Blasco, Ruth; Rosell, Jordi; González-Aguilera, Diego; Domínguez-Rodrigo, Manuel

    2017-09-01

    In the last few years, the study of cut marks on bone surfaces has become fundamental for the interpretation of prehistoric butchery practices. Due to the difficulties in the correct identification of cut marks, many criteria for their description and classification have been suggested. Different techniques, such as three-dimensional digital microscope (3D DM), laser scanning confocal microscopy (LSCM) and micro-photogrammetry (M-PG) have been recently applied to the study of cut marks. Although the 3D DM and LSCM microscopic techniques are the most commonly used for the 3D identification of cut marks, M-PG has also proved to be very efficient and a low-cost method. M-PG is a noninvasive technique that allows the study of the cortical surface without any previous preparation of the samples, and that generates high-resolution models. Despite the current application of microscopic and micro-photogrammetric techniques to taphonomy, their reliability has never been tested. In this paper, we compare 3D DM, LSCM and M-PG in order to assess their resolution and results. In this study, we analyse 26 experimental cut marks generated with a metal knife. The quantitative and qualitative information registered is analysed by means of standard multivariate statistics and geometric morphometrics to assess the similarities and differences obtained with the different methodologies. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  7. Biofilm formation on the Provox ActiValve: Composition and ingrowth analyzed by Illumina paired-end RNA sequencing, fluorescence in situ hybridization, and confocal laser scanning microscopy.

    Science.gov (United States)

    Timmermans, Adriana J; Harmsen, Hermie J M; Bus-Spoor, Carien; Buijssen, Kevin J D A; van As-Brooks, Corina; de Goffau, Marcus C; Tonk, Rudi H; van den Brekel, Michiel W M; Hilgers, Frans J M; van der Laan, Bernard F A M

    2016-04-01

    The most frequent cause of voice prosthesis failure is microbial biofilm formation on the silicone valve, leading to destruction of the material and transprosthetic leakage. The Provox ActiValve valve is made of fluoroplastic, which should be insusceptible to destruction. The purpose of this study was to determine if fluoroplastic is insusceptible to destruction by Candida species. Thirty-three dysfunctional Provox ActiValves (collected 2011-2013). Biofilm analysis was performed with Illumina paired-end sequencing (IPES), assessment of biofilm-material interaction with fluorescence in situ hybridization (FISH), and confocal laser scanning microscopy (CLSM). IPES (n = 10) showed that Candida albicans and Candida tropicalis are dominant populations on fluoroplastic and silicone. Microbial diversity is significantly lower on fluoroplastic. Lactobacillus gasseri is the prevalent bacterial strain on most voice prostheses. FISH and CLSM (n = 23): in none of the cases was ingrowth of Candida species present in the fluoroplastic. Fluoroplastic material of Provox ActiValve seems insusceptible to destruction by Candida species, which could help improve durability of voice prostheses. © 2015 Wiley Periodicals, Inc. Head Neck 38: E432-E440, 2016. © 2015 Wiley Periodicals, Inc.

  8. Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions.

    Science.gov (United States)

    Valle, Edith R; Henderson, Gemma; Janssen, Peter H; Cox, Faith; Alexander, Trevor W; McAllister, Tim A

    2015-06-01

    In this study, methanogen-specific coenzyme F420 autofluorescence and confocal laser scanning microscopy were used to identify rumen methanogens and define their spatial distribution in free-living, biofilm-, and protozoa-associated microenvironments. Fluorescence in situ hybridization (FISH) with temperature-controlled hybridization was used in an attempt to describe methanogen diversity. A heat pretreatment (65 °C, 1 h) was found to be a noninvasive method to increase probe access to methanogen RNA targets. Despite efforts to optimize FISH, 16S rRNA methanogen-specific probes, including Arch915, bound to some cells that lacked F420, possibly identifying uncharacterized Methanomassiliicoccales or reflecting nonspecific binding to other members of the rumen bacterial community. A probe targeting RNA from the methanogenesis-specific methyl coenzyme M reductase (mcr) gene was shown to detect cultured Methanosarcina cells with signal intensities comparable to those of 16S rRNA probes. However, the probe failed to hybridize with the majority of F420-emitting rumen methanogens, possibly because of differences in cell wall permeability among methanogen species. Methanogens were shown to integrate into microbial biofilms and to exist as ecto- and endosymbionts with rumen protozoa. Characterizing rumen methanogens and defining their spatial distribution may provide insight into mitigation strategies for ruminal methanogenesis.

  9. In situ detection of the Zn(2+) release process of ZnO NPs in tumour cells by confocal laser scanning fluorescence microscopy.

    Science.gov (United States)

    Song, Wenshuang; Tang, Xiaoling; Li, Yong; Sun, Yang; Kong, Jilie; Qingguang, Ren

    2016-08-01

    The use of zinc oxide (ZnO) nanoparticles (NPs) for cancer is not yet clear for human clinical applications, which is primarily due to the lack of a better understanding of the action mechanisms and cellular consequences of the direct exposure of cells to these NPs. In this work, the authors have selected zinquin ethyl ester, a Zn(2+)-specific fluorescent molecular probe, to efficiently differentiate ZnO NPs and Zn(2+), and combined with confocal laser scanning microscopy (CLSM) to in situ study the Zn(2+) release process of ZnO NPs in cancer cell system through detecting the change of Zn(2+) level over time. During the experiments, the authors have designed the test group ZnO-2 in addition to assess the influence of a long-term storage on the characteristics of ZnO NPs in aqueous solution, and the Zn(2+) release process of ZnO NPs in cancer cell system. After three-month storage at room temperature, the release process became earlier and faster, which was consistent with previous results of transmission electron microscope, UV-Vis and PL spectra. It is a good detection method that combination of Zn(2+)-specific fluorescent molecular probe and CLSM, which will be helpful for ZnO NPs using in clinical research.

  10. The development of exo-erythrocytic schizonts of Plasmodium berghei in vitro from gamma-irradiated and non-irradiated sporozoites: a study using confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Sinden, E.; Couchman, A.; Suhrbier, A.; Marsh, F.; Winger, L.; Ranawaka, G.

    1991-01-01

    Confocal scanning laser microscopy has been used to study the distribution of antigens expressed by the liver stages of Plasmodium berghei in cultured hepatoma cells. The 3-dimensional images obtained of intact parasites clearly show complex patterns of antigen expression not apparent when using conventional IFAT or immunoelectron microscopy. A liver-stage specific antigen (Pbl 1) was shown to be confined to the parasitophorous vacuole; the vacuole has extensive diverticulae extending into the host cell. Small parasites were detected for the first time in 'mature' cultures. These did not represent a distinct population, but the 'tail' of a broad continuum of parasite sizes. Irradiated sporozoites produce a transient population of slow-growing parasites which express a very limited range of antigens de novo in the invaded hepatoma cell. A comparison of the reactivity of normal EE parasites with anti-circumsporozoite antibody and with ant-Pbl 1 suggests that the former reagent may reliably be used to identify sporozoites invading host cells, but should not be used to determine the number of parasites that successfully undergo intrahepatic development. Anti-Pbl-1 indicates on 33% of invaded sporozoites identified by anti-CSP subsequently differentiate. (author)

  11. Quantitative evaluation of experimental choroidal neovascularization by confocal scanning laser ophthalmoscopy: fluorescein angiogram parallels heparan sulfate proteoglycan expression

    Directory of Open Access Journals (Sweden)

    C.V. Regatieri

    2010-07-01

    Full Text Available The objective of the present study was to develop a quantitative method to evaluate laser-induced choroidal neovascularization (CNV in a rat model using Heidelberg Retina Angiograph 2 (HRA2 imaging. The expression of two heparan sulfate proteoglycans (HSPG related to inflammation and angiogenesis was also investigated. CNV lesions were induced with argon laser in 21 heterozygous Zucker rats and after three weeks a fluorescein angiogram and autofluorescence exams were performed using HRA2. The area and greatest linear dimension were measured by two observers not aware of the protocol. Bland-Altman plots showed agreement between the observers, suggesting that the technique was reproducible. After fluorescein angiogram, HSPG (perlecan and syndecan-4 were analyzed by real-time RT-PCR and immunohistochemistry. There was a significant increase in the expression of perlecan and syndecan-4 (P < 0.0001 in retinas bearing CNV lesions compared to control retinas. The expression of these two HSPG increased with increasing CNV area. Immunohistochemistry demonstrated that the rat retina damaged with laser shots presented increased expression of perlecan and syndecan-4. Moreover, we observed that the overexpression occurred in the outer layer of the retina, which is related to choroidal damage. It was possible to develop a standardized quantitative method to evaluate CNV in a rat model using HRA2. In addition, we presented data indicating that the expression of HSPG parallels the area of CNV lesion. The understanding of these events offers opportunities for studies of new therapeutic interventions targeting these HSPG.

  12. Intensive care unit environmental surfaces are contaminated by multidrug-resistant bacteria in biofilms: combined results of conventional culture, pyrosequencing, scanning electron microscopy, and confocal laser microscopy.

    Science.gov (United States)

    Hu, H; Johani, K; Gosbell, I B; Jacombs, A S W; Almatroudi, A; Whiteley, G S; Deva, A K; Jensen, S; Vickery, K

    2015-09-01

    Hospital-associated infections cause considerable morbidity and mortality, and are expensive to treat. Organisms causing these infections can be sourced from the inanimate environment around a patient. Could the difficulty in eradicating these organisms from the environment be because they reside in dry surface biofilms? The intensive care unit (ICU) of a tertiary referral hospital was decommissioned and the opportunity to destructively sample clinical surfaces was taken in order to investigate whether multidrug-resistant organisms (MDROs) had survived the decommissioning process and whether they were present in biofilms. The ICU had two 'terminal cleans' with 500 ppm free chlorine solution; items from bedding, surrounds, and furnishings were then sampled with cutting implements. Sections were sonicated in tryptone soya broth and inoculated on to chromogenic plates to demonstrate MDROs, which were confirmed with the Vitek2 system. Genomic DNA was extracted directly from ICU samples, and subjected to polymerase chain reaction (PCR) for femA to detect Staphylococcus aureus and the microbiome by bacterial tag-encoded FLX amplicon pyrosequencing. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were performed on environmental samples. Multidrug-resistant bacteria were cultured from 52% (23/44) of samples cultured. S. aureus PCR was positive in 50%. Biofilm was demonstrated in 93% (41/44) of samples by CLSM and/or SEM. Pyrosequencing demonstrated that the biofilms were polymicrobial and contained species that had multidrug-resistant strains. Dry surface biofilms containing MDROs are found on ICU surfaces despite terminal cleaning with chlorine solution. How these arise and how they might be removed requires further study. Copyright © 2015 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

  13. Confocal Laser Endomicroscopy in Inflammatory Bowel Disease

    DEFF Research Database (Denmark)

    Rasmussen, Ditlev Nytoft; Karstensen, John Gásdal; Riis, Lene Buhl

    2015-01-01

    included. Next, eligible studies were analysed with respect to several parameters, such as technique and clinical aim and definitions of outcomes. RESULTS: Confocal laser endomicroscopy has been used for a wide range of purposes in inflammatory bowel disease, covering assessment of inflammatory severity...... of confocal laser endomicroscopy for inflammatory bowel disease. METHODS: Available literature was searched systematically for studies applying confocal laser endomicroscopy in Crohn's disease or ulcerative colitis. Relevant literature was reviewed and only studies reporting original clinical data were...... of histological features such as colonic crypts, epithelial gaps and epithelial leakiness to fluorescein. CONCLUSIONS: Confocal laser endomicroscopy remains an experimental but emerging tool for assessment of inflammatory bowel disease. It is the only method that enables in vivo functional assessment...

  14. Assessment of tissue fibrosis in skin biopsies from patients with systemic sclerosis employing confocal laser scanning microscopy: an objective outcome measure for clinical trials?

    Science.gov (United States)

    Busquets, Joanna; Del Galdo, Francesco; Kissin, Eugene Y.

    2010-01-01

    Objectives. To obtain an objective, unbiased assessment of skin fibrosis in patients with SSc for use in clinical trials of SSc disease-modifying therapeutics. Methods. Skin biopsies from the dorsal forearm of six patients with diffuse SSc and six healthy controls, and skin biopsies from the forearm of one patient with diffuse SSc before and following 1 year treatment with mycophenolate mofetil were analysed by confocal laser scanning microscopy (CLSM) with specific antibodies against collagen types I and III or fibronectin. The integrated density of fluorescence (IDF) was calculated employing National Institutes of Health-ImageJ software in at least four different fields per biopsy spanning the full dermal thickness. Results. The intensities of collagen types I and III and fibronectin IDF were 174, 147 and 139% higher in SSc skin than in normal skin, respectively. All differences were statistically significant. The sum of the IDF values obtained for the three proteins yielded a comprehensive fibrosis score. The average fibrosis score for the six SSc samples was 28.3 × 106 compared with 18.6 × 106 for the six normal skin samples (P < 0.0001). Comparison of skin biopsies obtained from the same SSc patient before treatment and after 12 months of treatment with mycophenolate mofetil showed a reduction of 39% in total fibrosis score after treatment. Conclusions. CLSM followed by quantitative image analysis provides an objective and unbiased assessment of skin fibrosis in SSc and could be a useful end-point for clinical trials with disease-modifying agents to monitor the response or progression of the disease. PMID:20202926

  15. In situ observation of the growth of biofouling layer in osmotic membrane bioreactors by multiple fluorescence labeling and confocal laser scanning microscopy.

    Science.gov (United States)

    Yuan, Bo; Wang, Xinhua; Tang, Chuyang; Li, Xiufen; Yu, Guanghui

    2015-05-15

    Since the concept of the osmotic membrane bioreactor (OMBR) was introduced in 2008, it has attracted growing interests for its potential applications in wastewater treatment and reclamation; however, the fouling mechanisms of forward osmosis (FO) membrane especially the development of biofouling layer in the OMBR are not yet clear. Here, the fouled FO membranes were obtained from the OMBRs on days 3, 8 and 25 in sequence, and then the structure and growing rule of the biofouling layer formed on the FO membrane samples were in-situ characterized by multiple fluorescence labeling and confocal laser scanning microscopy (CLSM). CLSM images indicated that the variations in abundance and distribution of polysaccharides, proteins and microorganisms in the biofouling layer during the operation of OMBRs were significantly different. Before the 8th day, their biovolume dramatically increased. Subsequently, the biovolumes of β-d-glucopyranose polysaccharides and proteins continued increasing and leveled off after 8 days, respectively, while the biovolumes of α-d-glucopyranose polysaccharides and microorganisms decreased. Extracellular polymeric substances (EPS) played a significant role in the formation and growth of biofouling layer, while the microorganisms were seldom detected on the upper fouling layer after 3 days. Based on the results obtained in this study, the growth of biofouling layer on the FO membrane surface in the OMBR could be divided into three stages. Initially, EPS was firstly deposited on the FO membrane surface, and then microorganisms associated with EPS located in the initial depositing layer to form clusters. After that, the dramatic increase of the clusters of EPS and microorganisms resulted in the quick growth of biofouling layer during the flux decline of the OMBR. However, when the water flux became stable in the OMBR, some microorganisms and EPS would be detached from the FO membrane surface. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

    NARCIS (Netherlands)

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

    2017-01-01

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial

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

    NARCIS (Netherlands)

    De Luca, G.; Breedijk, R.; Hoebe, R.; Stallinga, S.; Manders, E.

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial

  18. Fluorescence excitation analysis by two-photon confocal laser scanning microscopy: a new method to identify fluorescent nanoparticles on histological tissue sections

    Directory of Open Access Journals (Sweden)

    Kahn E

    2012-10-01

    Full Text Available Edmond Kahn,1 Nicolas Tissot,3 Perrine Frere,3 Aurélien Dauphin,3 Mohamed Boumhras,2,4 Claude-Marie Bachelet,3 Frédérique Frouin,1 Gérard Lizard21Institut National de la Santé et de la Recherche Médicale (INSERM U678/UMR-S UPMC, CHU Pitié-Salpêtrière, Paris, France; 2Equipe Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique EA7270, Faculté des Sciences Gabriel, Université de Bourgogne-INSERM Dijon, France; 3Plateforme d'Imagerie cellulaire, UPMC, Paris, France; 4Laboratory of Biochemistry and Neuroscience, Applied Toxicology Group, Faculty of Science and Technology, Settat, MoroccoAbstract: In the present study, we make use of the ability of two-photon confocal laser scanning microscopes (CLSMs equipped with tunable lasers to produce spectral excitation image sequences. Furthermore, unmixing, which is usually performed on emission image sequences, is performed on these excitation image sequences. We use factor analysis of medical image sequences (FAMIS, which produces factor images, to unmix spectral image sequences of stained structures in tissue sections to provide images of characterized stained cellular structures. This new approach is applied to histological tissue sections of mouse aorta containing labeled iron nanoparticles stained with Texas Red and counterstained with SYTO13, to obtain visual information about the accumulation of these nanoparticles in the arterial wall. The possible presence of Texas Red is determined using a two-photon CLSM associated with FAMIS via the excitation spectra. Texas Red and SYTO13 are thus differentiated, and corresponding factor images specify their possible presence and cellular localization. In conclusion, the designed protocol shows that sequences of images obtained by excitation in a two-photon CLSM enables characterization of Texas Red-stained nanoparticles and other markers. This methodology offers an alternative and complementary solution to the conventional use of emission

  19. Confocal laser endomicroscopy in ulcerative colitis

    DEFF Research Database (Denmark)

    Karstensen, John Gásdal; Săftoiu, Adrian; Brynskov, Jørn

    2016-01-01

    BACKGROUND AND AIMS: Confocal laser endomicroscopy enables real-time in vivo microscopy during endoscopy and can predict relapse in patients with inflammatory bowel disease in remission. However, little is known about how endomicroscopic features change with time. The aim of this longitudinal study...... was to correlate colonic confocal laser endomicroscopy (CLE) in ulcerative colitis with histopathology and macroscopic appearance before and after intensification of medical treatment. METHODS: Twenty-two patients with ulcerative colitis in clinical relapse and 7 control subjects referred for colonoscopy were...

  20. Integrated Confocal and Scanning Probe Microscopy for Biomedical Research

    Directory of Open Access Journals (Sweden)

    B.J. Haupt

    2006-01-01

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

  1. A fluorescence in situ staining method for investigating spores and vegetative cells of Clostridia by confocal laser scanning microscopy and structured illuminated microscopy.

    Science.gov (United States)

    D'Incecco, P; Ong, L; Gras, S; Pellegrino, L

    2018-04-18

    Non-pathogenic spore-forming Clostridia are of increasing interest due to their application in biogas production and their capability to spoil different food products. The life cycle for Clostridium includes a spore stage that can assist in survival under environmentally stressful conditions, such as extremes of temperature or pH. Due to their size, spores can be investigated by a range of microscopic techniques, many of which involve sample pre-treatment. We have developed a quick, simple and non-destructive fluorescent staining procedure that allows a clear differentiation between spores and vegetative cells and effectively stains spores, allowing recovery and tracking in subsequent experiments. Hoechst 34580, Propidium iodide and wheat germ agglutinin WGA 488 were used in combination to stain four strains of Clostridia at different life cycle stages. Staining was conducted without drying the sample, preventing changes induced by dehydration and cells observed by confocal laser scanner microscopy or using a super-resolution microscope equipped with a 3D-structured illumination module. Dual staining with Hoechst/Propidium iodide differentiated spores from vegetative cells, provided information on the viability of cells and was successfully applied to follow spore production induced by heating. Super-resolution microscopy of spores probed by Hoechst 34580 also allowed chromatin to be visualised. Direct staining of a cheese specimen using Nile Red and Fast Green allowed in situ observation of spores within the cheese and their position within the cheese matrix. The proposed staining method has broad applicability and can potentially be applied to follow Clostridium spore behaviour in a range of different environments. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Sub-Airy Confocal Adaptive Optics Scanning Ophthalmoscopy.

    Science.gov (United States)

    Sredar, Nripun; Fagbemi, Oladipo E; Dubra, Alfredo

    2018-04-01

    To demonstrate the viability of improving transverse image resolution in reflectance scanning adaptive optics ophthalmoscopy using sub-Airy disk confocal detection. The foveal cone mosaic was imaged in five human subjects free of known eye disease using two custom adaptive optics scanning light ophthalmoscopes (AOSLOs) in reflectance with 7.75 and 4.30 mm pupil diameters. Confocal pinholes of 0.5, 0.6, 0.8, and 1.0 Airy disk diameters (ADDs) were used in a retinal conjugate plane before the light detector. Average cone photoreceptor intensity profile width and power spectrum were calculated for the resulting images. Detected energy using a model eye was recorded for each pinhole size. The cone photoreceptor mosaic is better resolved with decreasing confocal pinhole size, with the high spatial frequency content of the images enhanced in both the large- and small-pupil AOSLOs. The average cone intensity profile width was reduced by ∼15% with the use of a 0.5 ADD pinhole when compared to a 1.0 ADD, with an accompanying reduction in signal greater than a factor of four. The use of sub-Airy disk confocal pinhole detection without increasing retinal light exposure results in a substantial improvement in image resolution at the cost of larger than predicted signal reduction. Improvement in transverse resolution using sub-Airy disk confocal detection is a practical and low-cost approach that is applicable to all point- and line-scanning ophthalmoscopes, including optical coherence tomographers.

  3. Association of tongue brushing with the number of fungiform taste buds and taste perception: A preliminary study using confocal laser scanning microscopy in combination with a filter-paper disc method.

    Science.gov (United States)

    Kobayashi, Junichi; Saito, Takehisa; Ito, Tetsufumi; Yoshimura, Hitoshi; Matsuda, Shinpei; Yoshida, Hisato; Fujita, Ryousuke; Sano, Kazuo

    2017-12-01

    The aim of this study was to investigate the association of tongue brushing with the number of fungiform taste buds and taste perception using a confocal laser scanning microscopy in combination with a filter-paper disc method (FPDM). Twenty-four subjects with or without a habit of tongue brushing (11 males and 13 females, 20-46 years old) participated in this study. Nine of the 24 subjects had no habit of tongue brushing (Group 1, n=9). Fifteen subjects had a habit of tongue brushing, and the brushing regions of the tongue were as follows: central region (Group 2, n=7), or entire region (Group 3, n=8) of the tongue dorsum. Using confocal laser scanning microscopy, the average number of taste buds per fungiform papilla (FP) was counted. Taste perception was evaluated using an FPDM. These observations were performed in the midlateral region of the tongue since the distribution of fungiform papillae is large in the midlateral region compared to that in the central region. The subjects in Group 3 showed a significantly decreased number of fungiform taste buds compared to Group 1 and Group 2. Group 3 also showed significantly higher FPDM scores than the other two groups. Excessive tongue brushing of the entire tongue dorsum, including the midlateral region, may have an association with the decreased number of FP and taste buds and decreased taste sensation. To avoid these conditions, instituting proper tongue brushing methods, such as limiting it to the central region of the tongue and using a light touch, is suggested and is important for the subjects who are eager to participate in tongue brushing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Hyperchromatic laser scanning cytometry

    Science.gov (United States)

    Tárnok, Attila; Mittag, Anja

    2007-02-01

    In the emerging fields of high-content and high-throughput single cell analysis for Systems Biology and Cytomics multi- and polychromatic analysis of biological specimens has become increasingly important. Combining different technologies and staining methods polychromatic analysis (i.e. using 8 or more fluorescent colors at a time) can be pushed forward to measure anything stainable in a cell, an approach termed hyperchromatic cytometry. For cytometric cell analysis microscope based Slide Based Cytometry (SBC) technologies are ideal as, unlike flow cytometry, they are non-consumptive, i.e. the analyzed sample is fixed on the slide. Based on the feature of relocation identical cells can be subsequently reanalyzed. In this manner data on the single cell level after manipulation steps can be collected. In this overview various components for hyperchromatic cytometry are demonstrated for a SBC instrument, the Laser Scanning Cytometer (Compucyte Corp., Cambridge, MA): 1) polychromatic cytometry, 2) iterative restaining (using the same fluorochrome for restaining and subsequent reanalysis), 3) differential photobleaching (differentiating fluorochromes by their different photostability), 4) photoactivation (activating fluorescent nanoparticles or photocaged dyes), and 5) photodestruction (destruction of FRET dyes). With the intelligent combination of several of these techniques hyperchromatic cytometry allows to quantify and analyze virtually all components of relevance on the identical cell. The combination of high-throughput and high-content SBC analysis with high-resolution confocal imaging allows clear verification of phenotypically distinct subpopulations of cells with structural information. The information gained per specimen is only limited by the number of available antibodies and by sterical hindrance.

  5. Laser Scanning in Forests

    Directory of Open Access Journals (Sweden)

    Håkan Olsson

    2012-09-01

    Full Text Available The introduction of Airborne Laser Scanning (ALS to forests has been revolutionary during the last decade. This development was facilitated by combining earlier ranging lidar discoveries [1–5], with experience obtained from full-waveform ranging radar [6,7] to new airborne laser scanning systems which had components such as a GNSS receiver (Global Navigation Satellite System, IMU (Inertial Measurement Unit and a scanning mechanism. Since the first commercial ALS in 1994, new ALS-based forest inventory approaches have been reported feasible for operational activities [8–12]. ALS is currently operationally applied for stand level forest inventories, for example, in Nordic countries. In Finland alone, the adoption of ALS for forest data collection has led to an annual savings of around 20 M€/year, and the work is mainly done by companies instead of governmental organizations. In spite of the long implementation times and there being a limited tradition of making changes in the forest sector, laser scanning was commercially and operationally applied after about only one decade of research. When analyzing high-ranked journal papers from ISI Web of Science, the topic of laser scanning of forests has been the driving force for the whole laser scanning research society over the last decade. Thus, the topic “laser scanning in forests” has provided a significant industrial, societal and scientific impact. [...

  6. Evidence for highly localized damage in internal tin and powder-in-tube Nb{sub 3}Sn strands rolled before reaction obtained from coupled magneto-optical imaging and confocal laser scanning microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Polyanskii, A A; Lee, P J; Jewell, M C; Larbalestier, D C [Applied Superconductivity Center, National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310 (United States); Barzi, E; Turrioni, D; Zlobin, A V [Fermi National Accelerator Laboratory, Batavia, IL 60510 (United States)

    2009-09-15

    Nb{sub 3}Sn strands for high-current, high-field magnets must be cabled before reaction while the conductor is still composed of ductile components. Even though still in the ductile, deformable state, significant damage can occur in this step, which expresses itself by inhomogeneous A15 formation, Sn leakage or even worse effects during later reaction. In this study, we simulate cabling damage by rolling recent high performance powder-in-tube (PIT) and internal tin (IT) strands in controlled increments, applying standard Nb{sub 3}Sn reaction heat treatments, and then examining the local changes using magneto-optical imaging (MOI), scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). These combined characterizations allow any local damage to the filament architecture to be made clear. MOI directly reveals the local variation of superconductivity while CLSM is extremely sensitive in revealing Sn leakage beyond the diffusion barrier into the stabilizing Cu. These techniques reveal a markedly different response to deformation by the PIT and IT strands. The study demonstrates that these tools can provide a local, thorough, and detailed view of how strands degrade and thus complement more complex extracted strand studies.

  7. 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.

    Science.gov (United States)

    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.

  8. Scanning laser Doppler vibrometry

    DEFF Research Database (Denmark)

    Brøns, Marie; Thomsen, Jon Juel

    With a Scanning Laser Doppler Vibrometer (SLDV) a vibrating surface is automatically scanned over predefined grid points, and data processed for displaying vibration properties like mode shapes, natural frequencies, damping ratios, and operational deflection shapes. Our SLDV – a PSV-500H from...

  9. Fluorescence imaging of reactive oxygen species by confocal laser scanning microscopy for track analysis of synchrotron X-ray photoelectric nanoradiator dose: X-ray pump-optical probe.

    Science.gov (United States)

    Jeon, Jae Kun; Han, Sung Mi; Kim, Jong Ki

    2016-09-01

    Bursts of emissions of low-energy electrons, including interatomic Coulomb decay electrons and Auger electrons (0-1000 eV), as well as X-ray fluorescence produced by irradiation of large-Z element nanoparticles by either X-ray photons or high-energy ion beams, is referred to as the nanoradiator effect. In therapeutic applications, this effect can damage pathological tissues that selectively take up the nanoparticles. Herein, a new nanoradiator dosimetry method is presented that uses probes for reactive oxygen species (ROS) incorporated into three-dimensional gels, on which macrophages containing iron oxide nanoparticles (IONs) are attached. This method, together with site-specific irradiation of the intracellular nanoparticles from a microbeam of polychromatic synchrotron X-rays (5-14 keV), measures the range and distribution of OH radicals produced by X-ray emission or superoxide anions ({\\rm{O}}_2^-) produced by low-energy electrons. The measurements are based on confocal laser scanning of the fluorescence of the hydroxyl radical probe 2-[6-(4'-amino)phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF) or the superoxide probe hydroethidine-dihydroethidium (DHE) that was oxidized by each ROS, enabling tracking of the radiation dose emitted by the nanoradiator. In the range 70 µm below the irradiated cell, ^\\bullet{\\rm{OH}} radicals derived mostly from either incident X-ray or X-ray fluorescence of ION nanoradiators are distributed along the line of depth direction in ROS gel. In contrast, {\\rm{O}}_2^- derived from secondary electron or low-energy electron emission by ION nanoradiators are scattered over the ROS gel. ROS fluorescence due to the ION nanoradiators was observed continuously to a depth of 1.5 mm for both oxidized APF and oxidized DHE with relatively large intensity compared with the fluorescence caused by the ROS produced solely by incident primary X-rays, which was limited to a depth of 600 µm, suggesting dose enhancement as well as more

  10. Exploring the diversity of Listeria monocytogenes biofilm architecture by high-throughput confocal laser scanning microscopy and the predominance of the honeycomb-like morphotype.

    Science.gov (United States)

    Guilbaud, Morgan; Piveteau, Pascal; Desvaux, Mickaël; Brisse, Sylvain; Briandet, Romain

    2015-03-01

    Listeria monocytogenes is involved in food-borne illness with a high mortality rate. The persistence of the pathogen along the food chain can be associated with its ability to form biofilms on inert surfaces. While most of the phenotypes associated with biofilms are related to their spatial organization, most published data comparing biofilm formation by L. monocytogenes isolates are based on the quantitative crystal violet assay, which does not give access to structural information. Using a high-throughput confocal-imaging approach, the aim of this work was to decipher the structural diversity of biofilms formed by 96 L. monocytogenes strains isolated from various environments. Prior to large-scale analysis, an experimental design was created to improve L. monocytogenes biofilm formation in microscopic-grade microplates, with special emphasis on the growth medium composition. Microscopic analysis of biofilms formed under the selected conditions by the 96 isolates revealed only weak correlation between the genetic lineages of the isolates and the structural properties of the biofilms. However, a gradient in their geometric descriptors (biovolume, mean thickness, and roughness), ranging from flat multilayers to complex honeycomb-like structures, was shown. The dominant honeycomb-like morphotype was characterized by hollow voids hosting free-swimming cells and localized pockets containing mixtures of dead cells and extracellular DNA (eDNA). Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  11. Configurations of the Re-scan Confocal Microscope (RCM) for biomedical applications

    NARCIS (Netherlands)

    de Luca, G. M. R.; Desclos, E.; Breedijk, R. M. P.; Dolz-Edo, L.; Smits, G. J.; Bielefeld, P.; Picavet, L.; Fitzsimons, C. P.; Hoebe, R.; Manders, E. M. M.

    2017-01-01

    The new high-sensitive and high-resolution technique, Re-scan Confocal Microscopy (RCM), is based on a standard confocal microscope extended with a re-scan detection unit. The re-scan unit includes a pair of re-scanning mirrors that project the emission light onto a camera in a scanning manner. The

  12. Configurations of the Re-scan Confocal Microscope (RCM) for biomedical applications

    NARCIS (Netherlands)

    De Luca, G.M.R.; Desclos, E.; Breedijk, R.M.P.; Dolz-Edo, L.; Smits, G.J.; Nahidiazar, L.; Bielefeld, P.; Picavet, L.; Fitzsimons, C.P.; Hoebe, R.; Manders, E.M.M.

    The new high-sensitive and high-resolution technique, Re-scan Confocal Microscopy (RCM), is based on a standard confocal microscope extended with a re-scan detection unit. The re-scan unit includes a pair of re-scanning mirrors that project the emission light onto a camera in a scanning manner. The

  13. 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.

    Science.gov (United States)

    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.

  14. Pigment organization effects on energy transfer and Chl a emission imaged in the diatoms C. meneghiniana and P. tricornutum in vivo: a confocal laser scanning fluorescence (CLSF) microscopy and spectroscopy study.

    Science.gov (United States)

    Premvardhan, Lavanya; Réfrégiers, Matthieu; Büchel, Claudia

    2013-09-26

    The (auto)fluorescence from three diatom strains, Cyclotella meneghiniana (Cm), Phaeodactylum tricornutum 1a (Pt1a), and Phaeodactylum UTex (PtUTex), has been imaged in vivo to submicrometer resolution using confocal laser scanning fluorescence (CLSF) microscopy. The diatoms are excited at 473 and 532 nm, energy primarily absorbed by the carotenoid fucoxanthin (Fx) found within the fucoxanthin chlorophyll a/c proteins (FCPs). On the basis of the fluorescence spectra measured in each image voxel, we obtain information about the spatial and energetic distribution of the terminal Chl a emitters, localized in the FCPs and the reaction centers of the PSII protein complexes, and the nature and location of the primary absorbers that are linked to these emitters; 532 nm excites the highly efficient Fx(red) light harvesters, and lesser amounts of Fx(green)s, that are enriched in some FCPs and preferentially transfer energy to PSII, compared to 473 nm, which excites almost equal amounts of all three previously identified sets of Fx--Fx(red), Fx(green) and Fx(blue)--as well as Chl c. The heterogeneous Chl a emission observed from the (C)LSF images indicates that the different Fx's serve different final emitters in P. tricornutum and suggest, at least in C. meneghiniana , a localization of FCPs with relatively greater Fx(red) content at the chloroplast edges, but with overall higher FCP concentration in the interior of the plastid. To better understand our results, the concentration-dependent ensemble-averaged diatom solution spectra are compared to the (auto)fluorescence spectra of individual diatoms, which indicate that pigment packing effects at an intracellular level do affect the diatoms' spectral properties, in particular, concerning a 710 nm emission band apparent under stress conditions. A species-specific response of the spectral signature to the incident light is also discussed in terms of the presence of a silica shell in Cm but not in Pt1a nor PtUTex.

  15. Extended Field Laser Confocal Microscopy (EFLCM): Combining automated Gigapixel image capture with in silico virtual microscopy

    International Nuclear Information System (INIS)

    Flaberg, Emilie; Sabelström, Per; Strandh, Christer; Szekely, Laszlo

    2008-01-01

    Confocal laser scanning microscopy has revolutionized cell biology. However, the technique has major limitations in speed and sensitivity due to the fact that a single laser beam scans the sample, allowing only a few microseconds signal collection for each pixel. This limitation has been overcome by the introduction of parallel beam illumination techniques in combination with cold CCD camera based image capture. Using the combination of microlens enhanced Nipkow spinning disc confocal illumination together with fully automated image capture and large scale in silico image processing we have developed a system allowing the acquisition, presentation and analysis of maximum resolution confocal panorama images of several Gigapixel size. We call the method Extended Field Laser Confocal Microscopy (EFLCM). We show using the EFLCM technique that it is possible to create a continuous confocal multi-colour mosaic from thousands of individually captured images. EFLCM can digitize and analyze histological slides, sections of entire rodent organ and full size embryos. It can also record hundreds of thousands cultured cells at multiple wavelength in single event or time-lapse fashion on fixed slides, in live cell imaging chambers or microtiter plates. The observer independent image capture of EFLCM allows quantitative measurements of fluorescence intensities and morphological parameters on a large number of cells. EFLCM therefore bridges the gap between the mainly illustrative fluorescence microscopy and purely quantitative flow cytometry. EFLCM can also be used as high content analysis (HCA) instrument for automated screening processes

  16. UNC Pembroke Laser Scanning Confocal Microscopy Facility

    Science.gov (United States)

    2016-04-29

    an aggregation-dependent manner. Biochim Biophys Acta (Mol. Basis of Disease) 1812:1664-1674. Technology Transfer 1    Scientific progress and...receptors and presynaptic markers in an aggregation-dependent manner. Biochim Biophys Acta (Mol. Basis of Disease) 1812:1664-1674.

  17. Ultrasensitive and selective detection of mercury (II) in serum based on the gold film sensor using a laser scanning confocal imaging-surface plasmon resonance system in real time

    Science.gov (United States)

    Liu, Sha; Zhang, Hongyan; Liu, Weimin; Wang, Pengfei

    2015-10-01

    Hg2+ ions are one of the most toxic heavy metal ion pollutants, and are caustic and carcinogenic materials with high cellular toxicity. The Hg2+ ions can accumulate in the human body through the food chain and cause serious and permanent damage to the brain with both acute and chronic toxicity. According to the US Environment Protection Agency (EPA) guidelines, Hg2+ ions must be at concentrations below 1 ng/ml (10 nM) in drinking water. If the Hg2+ ions are higher than 2.5 ng/ml in serum, that will bring mercury poisoning. The traditional testing for Hg2+ ions includes atomic absorption, atomic fluorescence, and inductively coupled plasma mass spectrometry. These methods are usually coupled with gas chromatography, high-performance liquid chromatography, and capillary electrophoresis. However, these instrument-based techniques are rather complicated, time-consuming, costly, and unsuitable for online and portable use. An ultrasensitive and selective detection of mercury (II) in serum was investigated using a laser scanning confocal imaging-surface plasmon resonance system (LSCI-SPR). The detection limit was as low as 0.01 ng/ml for Hg2+ ions in fetal calf serum and that is lower than that was required Hg2+ ions must be at concentrations below 1 ng/ml by the US Environment Protection Agency (EPA) guidelines. This sensor was designed on a T-rich, single-stranded DNA (ssDNA)-modified gold film, which can be individually manipulated using specific T-Hg2+-T complex formation. The quenching intensity of the fluorescence images for rhodamine-labeled ssDNA fitted well with the changes in SPR. The changes varied with the Hg2+ ion concentration, which is unaffected by the presence of other metal ions. A good liner relation was got with the coefficients of 0.9116 in 30% fetal calf serums with the linear part over a range of 0.01 ng/ml to10 ng/ml.

  18. Evaluating the Toxicity/Fixation Balance for Corneal Cross-Linking With Sodium Hydroxymethylglycinate (SMG) and Riboflavin-UVA (CXL) in an Ex Vivo Rabbit Model Using Confocal Laser Scanning Fluorescence Microscopy.

    Science.gov (United States)

    Kim, Su-Young; Babar, Natasha; Munteanu, Emilia Laura; Takaoka, Anna; Zyablitskaya, Mariya; Nagasaki, Takayuki; Trokel, Stephen L; Paik, David C

    2016-04-01

    To develop methods to delineate the relationship between endothelial cell toxicity and tissue fixation (toxicity/fixation) using sodium hydroxymethylglycinate (SMG), a formaldehyde releaser, and riboflavin-UVA photochemical corneal cross-linking (CXL) for therapeutic tissue cross-linking of the cornea. Eleven fresh cadaveric rabbit heads were used for ex vivo corneal cross-linking simulation. After epithelial debridement, the tissue was exposed to 1/4 max (9.8 mM) or 1/3 max (13 mM) SMG at pH 8.5 for 30 minutes or riboflavin-UVA (CXL). The contralateral cornea served as a paired control. Postexposure, cross-linking efficacy was determined by thermal denaturation temperature (Tm) and endothelial damage was assessed using calcein AM and ethidium homodimer staining (The Live/Dead Kit). Confocal laser scanning fluorescence microscopy was used to generate live/dead cell counts using a standardized algorithm. The ΔTm after CXL, 1/3 SMG, and 1/4 SMG was 2.2 ± 0.9°C, 1.3 ± 0.5°C, and 1.1 ± 0.5°C, respectively. Endothelial cell damage was expressed as the percent of dead cells/live + dead cells counted per high-power field. The values were 3 ± 1.7% (control) and 8.9 ± 11.1% (CXL) (P = 0.390); 1 ± 0.2% (control) and 19.5 ± 32.2% (1/3 max SMG) (P = 0.426); and 2.7 ± 2.4% (control) and 2.8 ± 2.2% (1/4 max SMG) (P = 0.938). The values for endothelial toxicity were then indexed over the shift in Tm to yield a toxicity/fixation index. The values were as follows: 2.7 for CXL, 14 for 1/3 max, and 0.1 for 1/4 max. Quarter max (1/4 max = 9.8 mM) SMG effectively cross-linked tissue and was nontoxic to endothelial cells. Thus, SMG is potentially a compound that could achieve both desired effects.

  19. Confocal scanning microscopy with multiple optical probes for high speed measurements and better imaging

    Science.gov (United States)

    Chun, Wanhee; Lee, SeungWoo; Gweon, Dae-Gab

    2008-02-01

    Confocal scanning microscopy (CSM) needs a scanning mechanism because only one point information of specimen can be obtained. Therefore the speed of the confocal scanning microscopy is limited by the speed of the scanning tool. To overcome this limitation from scanning tool we propose another scanning mechanism. We make three optical probes in the specimen under confocal condition of each point. Three optical probes are moved by beam scanning mechanism with shared resonant scanning mirror (RM) and galvanometer driven mirror (GM). As each optical probe scan allocated region of the specimen, information from three points is obtained simultaneously and image acquisition time is reduced. Therefore confocal scanning microscopy with multiple optical probes is expected to have three times faster speed of the image acquisition than conventional one. And as another use, multiple optical probes to which different light wavelength is applied can scan whole same region respectively. It helps to obtain better contrast image in case of specimens having different optical characteristics for specific light wavelength. In conclusion confocal scanning microscopy with multiple optical probes is useful technique for views of image acquisition speed and image quality.

  20. Dual-detection confocal fluorescence microscopy: fluorescence axial imaging without axial scanning.

    Science.gov (United States)

    Lee, Dong-Ryoung; Kim, Young-Duk; Gweon, Dae-Gab; Yoo, Hongki

    2013-07-29

    We propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system. Axial information on fluorescent emitters can be measured by the axial response curve through the ratio of intensity signals. DDCFM can rapidly acquire a 3-D fluorescent image from a single two-dimensional scan with less phototoxicity and photobleaching than confocal fluorescence microscopy because no mechanical depth scans are needed. We demonstrated the feasibility of the proposed method by phantom studies.

  1. EUS-Guided Needle-Based Confocal Laser Endomicroscopy

    DEFF Research Database (Denmark)

    Bhutani, Manoop S; Koduru, Pramoda; Joshi, Virendra

    2015-01-01

    Endoscopic ultrasound (EUS) has emerged as an excellent tool for imaging the gastrointestinal tract, as well as surrounding structures. EUS-guided fine-needle aspiration (EUS-FNA) has become the standard of care for the tissue sampling of a variety of masses and lymph nodes within and around...... the gut, providing further diagnostic and staging information. Confocal laser endomicroscopy (CLE) is a novel endoscopic method that enables imaging at a subcellular level of resolution during endoscopy, allowing up to 1000-fold magnification of tissue and providing an optical biopsy. A new procedure...... that has been developed in the past few years is needle-based confocal laser endomicroscopy (nCLE), which involves a mini-CLE probe that can be passed through a 1 9-gauge needle during EUS-FNA. This enables the real-time visualization of tissue at a microscopic level, with the potential to further improve...

  2. Optical depth sectioning in the aberration-corrected scanning transmission and scanning confocal electron microscope

    International Nuclear Information System (INIS)

    Behan, G; Nellist, P D

    2008-01-01

    The use of spherical aberration correctors in the scanning transmission electron microscope (STEM) has the effect of reducing the depth of field of the microscope, making three-dimensional imaging of a specimen possible by optical sectioning. Depth resolution can be improved further by placing aberration correctors and lenses pre and post specimen to achieve an imaging mode known as scanning confocal electron microscopy (SCEM). We present the calculated incoherent point spread functions (PSF) and optical transfer functions (OTF) of a STEM and SCEM. The OTF for a STEM is shown to have a missing cone region which results in severe blurring along the optic axis, which can be especially severe for extended objects. We also present strategies for reconstruction of experimental data, such as three-dimensional deconvolution of the point spread function.

  3. Confocal laser feedback tomography for skin cancer detection.

    Science.gov (United States)

    Mowla, Alireza; Du, Benjamin Wensheng; Taimre, Thomas; Bertling, Karl; Wilson, Stephen; Soyer, H Peter; Rakić, Aleksandar D

    2017-09-01

    Tomographic imaging of soft tissue such as skin has a potential role in cancer detection. The penetration of infrared wavelengths makes a confocal approach based on laser feedback interferometry feasible. We present a compact system using a semiconductor laser as both transmitter and receiver. Numerical and physical models based on the known optical properties of keratinocyte cancers were developed. We validated the technique on three phantoms containing macro-structural changes in optical properties. Experimental results were in agreement with numerical simulations and structural changes were evident which would permit discrimination of healthy tissue and tumour. Furthermore, cancer type discrimination was also able to be visualized using this imaging technique.

  4. The Enhancement of 3D Scans Depth Resolution Obtained by Confocal Scanning of Porous Materials

    Science.gov (United States)

    Martisek, Dalibor; Prochazkova, Jana

    2017-12-01

    The 3D reconstruction of simple structured materials using a confocal microscope is widely used in many different areas including civil engineering. Nonetheless, scans of porous materials such as concrete or cement paste are highly problematic. The well-known problem of these scans is low depth resolution in comparison to the horizontal and vertical resolution. The degradation of the image depth resolution is caused by systematic errors and especially by different random events. Our method is focused on the elimination of such random events, mainly the additive noise. We use an averaging method based on the Lindeberg-Lévy theorem that improves the final depth resolution to a level comparable with horizontal and vertical resolution. Moreover, using the least square method, we also precisely determine the limit value of a depth resolution. Therefore, we can continuously evaluate the difference between current resolution and the optimal one. This substantially simplifies the scanning process because the operator can easily determine the required number of scans.

  5. The Enhancement of 3D Scans Depth Resolution Obtained by Confocal Scanning of Porous Materials

    Directory of Open Access Journals (Sweden)

    Martisek Dalibor

    2017-12-01

    Full Text Available The 3D reconstruction of simple structured materials using a confocal microscope is widely used in many different areas including civil engineering. Nonetheless, scans of porous materials such as concrete or cement paste are highly problematic. The well-known problem of these scans is low depth resolution in comparison to the horizontal and vertical resolution. The degradation of the image depth resolution is caused by systematic errors and especially by different random events. Our method is focused on the elimination of such random events, mainly the additive noise. We use an averaging method based on the Lindeberg-Lévy theorem that improves the final depth resolution to a level comparable with horizontal and vertical resolution. Moreover, using the least square method, we also precisely determine the limit value of a depth resolution. Therefore, we can continuously evaluate the difference between current resolution and the optimal one. This substantially simplifies the scanning process because the operator can easily determine the required number of scans.

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

    Science.gov (United States)

    Guo, Kaikai; Zheng, Guoan

    2017-03-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-06-15

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

  8. Probe-based confocal laser endomicroscopy (pCLE) - a new imaging technique for in situ localization of spermatozoa.

    Science.gov (United States)

    Trottmann, Matthias; Stepp, Herbert; Sroka, Ronald; Heide, Michael; Liedl, Bernhard; Reese, Sven; Becker, Armin J; Stief, Christian G; Kölle, Sabine

    2015-05-01

    In azoospermic patients, spermatozoa are routinely obtained by testicular sperm extraction (TESE). However, success rates of this technique are moderate, because the site of excision of testicular tissue is determined arbitrarily. Therefore the aim of this study was to establish probe-based laser endomicroscopy (pCLE) a noval biomedical imaging technique, which provides the opportunity of non-invasive, real-time visualisation of tissue at histological resolution. Using pCLE we clearly visualized longitudinal and horizontal views of the tubuli seminiferi contorti and localized vital spermatozoa. Obtained images and real-time videos were subsequently compared with confocal laser scanning microscopy (CLSM) of spermatozoa and tissues, respectively. Comparative visualization of single native Confocal laser scanning microscopy (CLSM, left) and probe-based laser endomicroscopy (pCLE, right) using Pro Flex(TM) UltraMini O after staining with acriflavine. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Enhancement of fluorescence confocal scanning microscopy lateral resolution by use of structured illumination

    International Nuclear Information System (INIS)

    Kim, Taejoong; Gweon, DaeGab; Lee, Jun-Hee

    2009-01-01

    Confocal microscopy is an optical imaging technique used to reconstruct three-dimensional images without physical sectioning. As with other optical microscopes, the lateral resolution of the confocal microscope cannot surpass the diffraction limit. This paper presents a novel imaging system, structured illumination confocal scanning microscopy (SICSM), that uses structured illumination to improve the lateral resolution of the confocal microscope. The SICSM can easily be implemented by introducing a structured illumination generating optics to conventional line-scanning fluorescence confocal microscopy. In this paper, we report our analysis of the lateral and axial resolutions of the SICSM by use of mathematical imaging theory. Numerical simulation results show that the lateral resolution of the SICSM is 1.43-fold better than that of the confocal microscope. In the axial direction, however, the resolution of the SICSM is ∼15% poorer than that of the confocal microscope. This deterioration arises because of a decrease in the axial cut-off frequency caused by the process of generating structured illumination. We propose the use of imaging conditions under which a compromise between the axial and lateral resolutions is chosen. Finally, we show simulated images of diversely shaped test objects to demonstrate the lateral and axial resolution performance of the SICSM

  10. Volumetry of human taste buds using laser scanning microscopy.

    Science.gov (United States)

    Just, T; Srur, E; Stachs, O; Pau, H W

    2009-10-01

    In vivo laser scanning confocal microscopy is a relatively new, non-invasive method for assessment of oral cavity epithelia. The penetration depth of approximately 200-400 microm allows visualisation of fungiform papillae and their taste buds. This paper describes the technique of in vivo volumetry of human taste buds. Confocal laser scanning microscopy used a diode laser at 670 nm for illumination. Digital laser scanning confocal microscopy equipment consisted of the Heidelberg Retina Tomograph HRTII and the Rostock Cornea Module. Volume scans of fungiform papillae were used for three-dimensional reconstruction of the taste bud. This technique supplied information on taste bud structure and enabled measurement and calculation of taste bud volume. Volumetric data from a 23-year-old man over a nine-day period showed only a small deviation in values. After three to four weeks, phenomenological changes in taste bud structures were found (i.e. a significant increase in volume, followed by disappearance of the taste bud and appearance of a new taste bud). The data obtained indicate the potential application of this non-invasive imaging modality: to evaluate variation of taste bud volume in human fungiform papillae with ageing; to study the effects of chorda tympani nerve transection on taste bud volume; and to demonstrate recovery of taste buds in patients with a severed chorda tympani nerve who show recovery of gustatory sensibility after surgery.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    KA. Al-Salihi

    2009-12-01

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

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

    Science.gov (United States)

    Yang, Yong-fa; Li, Qi

    2014-12-01

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

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

    Science.gov (United States)

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

    2017-02-01

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

  15. CCDiode: an optimal detector for laser confocal microscopes

    Science.gov (United States)

    Pawley, James B.; Blouke, Morley M.; Janesick, James R.

    1996-04-01

    The laser confocal microscope (LCM) is now an established research tool in biology and materials science. In biological applications, it is usually employed to detect the location of fluorescent market molecules and, under these conditions, signal levels from bright areas are often digitizer. To maintain the desired +/- 3 e noise level at the relatively high data rate of 1 MHz, our new device utilizes 64 separate readout amplifier/digitizer systems, operating in sequence. The resulting detector is more compact, efficient and reliable than the PMT it replaces but as its sensitive area is smaller than that of a PMT, it will require auxiliary optics when used with any LCM having a large (mm) pinhole. As the signal light is parallel, a simple lens mounted axially and with the CCDiode at its focus would suffice. Future versions may use 3 X 3 or 5 X 5 arrays of sensors to `track' the confocal spot as it is deflected by inhomogeneities of the specimen, change its effective size or shape or detect system misalignment.

  16. Post-PRK corneal scatter measurements with a scanning confocal slit photon counter

    Science.gov (United States)

    Taboada, John; Gaines, David; Perez, Mary A.; Waller, Steve G.; Ivan, Douglas J.; Baldwin, J. Bruce; LoRusso, Frank; Tutt, Ronald C.; Perez, Jose; Tredici, Thomas; Johnson, Dan A.

    2000-06-01

    Increased corneal light scatter or 'haze' has been associated with excimer laser photorefractive surgery of the cornea. The increased scatter can affect visual performance; however, topical steroid treatment post surgery substantially reduces the post PRK scatter. For the treatment and monitoring of the scattering characteristics of the cornea, various methods have been developed to objectively measure the magnitude of the scatter. These methods generally can measure scatter associated with clinically observable levels of haze. For patients with moderate to low PRK corrections receiving steroid treatment, measurement becomes fairly difficult as the haze clinical rating is non observable. The goal of this development was to realize an objective, non-invasive physical measurement that could produce a significant reading for any level including the background present in a normal cornea. As back-scatter is the only readily accessible observable, the instrument is based on this measurement. To achieve this end required the use of a confocal method to bias out the background light that would normally confound conventional methods. A number of subjects with nominal refractive errors in an Air Force study have undergone PRK surgery. A measurable increase in corneal scatter has been observed in these subjects whereas clinical ratings of the haze were noted as level zero. Other favorable aspects of this back-scatter based instrument include an optical capability to perform what is equivalent to an optical A-scan of the anterior chamber. Lens scatter can also be measured.

  17. Femtosecond laser subsurface scleral treatment in cadaver human sclera and evaluation using two-photon and confocal microscopy

    Science.gov (United States)

    Sun, Hui; Fan, Zhongwei; Yan, Ying; Lian, Fuqiang; Kurtz, Ron; Juhasz, Tibor

    2016-03-01

    Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial-thickness drainage channels can be created with femtosecond laser in the translucent sclera for the potential treatment of glaucoma. We demonstrate the creation of partial-thickness subsurface drainage channels with the femtosecond laser in the cadaver human eyeballs and describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. A femtosecond laser operating at a wavelength of 1700 nm was scanned along a rectangular raster pattern to create the partial thickness subsurface drainage channels in the sclera of cadaver human eyes. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such partial thickness subsurface scleral channels. Our studies suggest that the confocal and two-photon microscopy can be used to investigate femtosecond-laser created partial-thickness drainage channels in the sclera of cadaver human eyes.

  18. Confocal soft X-ray scanning transmission microscopy: setup, alignment procedure and limitations

    International Nuclear Information System (INIS)

    Späth, Andreas; Raabe, Jörg; Fink, Rainer H.

    2015-01-01

    A conventional STXM setup has been upgraded with a second micro zone plate and aligned to confocal geometry. Two confocal geometries (in-line and off-axis) have been evaluated and a discussion on prospects and limitations is presented. Zone-plate-based scanning transmission soft X-ray microspectroscopy (STXM) is a well established technique for high-contrast imaging of sufficiently transparent specimens (e.g. ultrathin biological tissues, polymer materials, archaeometric specimens or magnetic thin films) with spatial resolutions in the regime of 20 nm and high spectroscopic or chemical sensitivity. However, due to the relatively large depth of focus of zone plates, the resolution of STXM along the optical axis so far stays unambiguously behind for thicker X-ray transparent specimens. This challenge can be addressed by the implementation of a second zone plate in the detection pathway of the beam, resulting in a confocal arrangement. Within this paper a first proof-of-principle study for a confocal STXM (cSTXM) and an elaborate alignment procedure in transmission and fluorescence geometry are presented. Based on first confocal soft X-ray micrographs of well known specimens, the advantage and limitation of cSTXM as well as further development potentials for future applications are discussed

  19. Confocal soft X-ray scanning transmission microscopy: setup, alignment procedure and limitations

    Energy Technology Data Exchange (ETDEWEB)

    Späth, Andreas [Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Egerlandstraße 3, 91058 Erlangen (Germany); Raabe, Jörg [Paul Scherrer Institut, 5232 Villigen (Switzerland); Fink, Rainer H., E-mail: rainer.fink@fau.de [Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Egerlandstraße 3, 91058 Erlangen (Germany); Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Egerlandstraße 3, 91058 Erlangen (Germany)

    2015-01-01

    A conventional STXM setup has been upgraded with a second micro zone plate and aligned to confocal geometry. Two confocal geometries (in-line and off-axis) have been evaluated and a discussion on prospects and limitations is presented. Zone-plate-based scanning transmission soft X-ray microspectroscopy (STXM) is a well established technique for high-contrast imaging of sufficiently transparent specimens (e.g. ultrathin biological tissues, polymer materials, archaeometric specimens or magnetic thin films) with spatial resolutions in the regime of 20 nm and high spectroscopic or chemical sensitivity. However, due to the relatively large depth of focus of zone plates, the resolution of STXM along the optical axis so far stays unambiguously behind for thicker X-ray transparent specimens. This challenge can be addressed by the implementation of a second zone plate in the detection pathway of the beam, resulting in a confocal arrangement. Within this paper a first proof-of-principle study for a confocal STXM (cSTXM) and an elaborate alignment procedure in transmission and fluorescence geometry are presented. Based on first confocal soft X-ray micrographs of well known specimens, the advantage and limitation of cSTXM as well as further development potentials for future applications are discussed.

  20. Cement paste surface roughness analysis using coherence scanning interferometry and confocal microscopy

    International Nuclear Information System (INIS)

    Apedo, K.L.; Munzer, C.; He, H.; Montgomery, P.; Serres, N.; Fond, C.; Feugeas, F.

    2015-01-01

    Scanning electron microscopy and scanning probe microscopy have been used for several decades to better understand the microstructure of cementitious materials. Very limited work has been performed to date to study the roughness of cementitious materials by optical microscopy such as coherence scanning interferometry (CSI) and chromatic confocal sensing (CCS). The objective of this paper is to better understand how CSI can be used as a tool to analyze surface roughness and topography of cement pastes. Observations from a series of images acquired using this technique on both polished and unpolished samples are described. The results from CSI are compared with those from a STIL confocal microscopy technique (SCM). Comparison between both optical techniques demonstrates the ability of CSI to measure both polished and unpolished cement pastes. - Highlights: • Coherence scanning interferometry (CSI) was used to analyze cement paste surfaces. • The results from the CSI were compared with those from a confocal microscopy. • 3D roughness parameters were obtained using the window resizing method. • Polished and unpolished cement pastes were studied

  1. Cement paste surface roughness analysis using coherence scanning interferometry and confocal microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Apedo, K.L., E-mail: apedo@unistra.fr [ICube, Université de Strasbourg, CNRS, 2 rue Boussingault, 67000 Strasbourg (France); Munzer, C.; He, H. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France); Montgomery, P. [ICube, Université de Strasbourg, CNRS, 23 rue du Loess, 67037 Strasbourg (France); Serres, N. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France); Fond, C. [ICube, Université de Strasbourg, CNRS, 2 rue Boussingault, 67000 Strasbourg (France); Feugeas, F. [ICube, INSA de Strasbourg, CNRS, 24 Bld de la Victoire, 67084 Strasbourg (France)

    2015-02-15

    Scanning electron microscopy and scanning probe microscopy have been used for several decades to better understand the microstructure of cementitious materials. Very limited work has been performed to date to study the roughness of cementitious materials by optical microscopy such as coherence scanning interferometry (CSI) and chromatic confocal sensing (CCS). The objective of this paper is to better understand how CSI can be used as a tool to analyze surface roughness and topography of cement pastes. Observations from a series of images acquired using this technique on both polished and unpolished samples are described. The results from CSI are compared with those from a STIL confocal microscopy technique (SCM). Comparison between both optical techniques demonstrates the ability of CSI to measure both polished and unpolished cement pastes. - Highlights: • Coherence scanning interferometry (CSI) was used to analyze cement paste surfaces. • The results from the CSI were compared with those from a confocal microscopy. • 3D roughness parameters were obtained using the window resizing method. • Polished and unpolished cement pastes were studied.

  2. Ribbon scanning confocal for high-speed high-resolution volume imaging of brain.

    Directory of Open Access Journals (Sweden)

    Alan M Watson

    Full Text Available Whole-brain imaging is becoming a fundamental means of experimental insight; however, achieving subcellular resolution imagery in a reasonable time window has not been possible. We describe the first application of multicolor ribbon scanning confocal methods to collect high-resolution volume images of chemically cleared brains. We demonstrate that ribbon scanning collects images over ten times faster than conventional high speed confocal systems but with equivalent spectral and spatial resolution. Further, using this technology, we reconstruct large volumes of mouse brain infected with encephalitic alphaviruses and demonstrate that regions of the brain with abundant viral replication were inaccessible to vascular perfusion. This reveals that the destruction or collapse of large regions of brain micro vasculature may contribute to the severe disease caused by Venezuelan equine encephalitis virus. Visualization of this fundamental impact of infection would not be possible without sampling at subcellular resolution within large brain volumes.

  3. Handbook of optical and laser scanning

    CERN Document Server

    Marshall, Gerald F

    2011-01-01

    From its initial publication titled Laser Beam Scanning in 1985 to Handbook of Optical and Laser Scanning, now in its second edition, this reference has kept professionals and students at the forefront of optical scanning technology. Carefully and meticulously updated in each iteration, the book continues to be the most comprehensive scanning resource on the market. It examines the breadth and depth of subtopics in the field from a variety of perspectives. The Second Edition covers: Technologies such as piezoelectric devices Applications of laser scanning such as Ladar (laser radar) Underwater

  4. Toward real-time virtual biopsy of oral lesions using confocal laser endomicroscopy interfaced with embedded computing.

    Science.gov (United States)

    Thong, Patricia S P; Tandjung, Stephanus S; Movania, Muhammad Mobeen; Chiew, Wei-Ming; Olivo, Malini; Bhuvaneswari, Ramaswamy; Seah, Hock-Soon; Lin, Feng; Qian, Kemao; Soo, Khee-Chee

    2012-05-01

    Oral lesions are conventionally diagnosed using white light endoscopy and histopathology. This can pose a challenge because the lesions may be difficult to visualise under white light illumination. Confocal laser endomicroscopy can be used for confocal fluorescence imaging of surface and subsurface cellular and tissue structures. To move toward real-time "virtual" biopsy of oral lesions, we interfaced an embedded computing system to a confocal laser endomicroscope to achieve a prototype three-dimensional (3-D) fluorescence imaging system. A field-programmable gated array computing platform was programmed to enable synchronization of cross-sectional image grabbing and Z-depth scanning, automate the acquisition of confocal image stacks and perform volume rendering. Fluorescence imaging of the human and murine oral cavities was carried out using the fluorescent dyes fluorescein sodium and hypericin. Volume rendering of cellular and tissue structures from the oral cavity demonstrate the potential of the system for 3-D fluorescence visualization of the oral cavity in real-time. We aim toward achieving a real-time virtual biopsy technique that can complement current diagnostic techniques and aid in targeted biopsy for better clinical outcomes.

  5. Scanning laser polarimetry in glaucoma.

    Science.gov (United States)

    Dada, Tanuj; Sharma, Reetika; Angmo, Dewang; Sinha, Gautam; Bhartiya, Shibal; Mishra, Sanjay K; Panda, Anita; Sihota, Ramanjit

    2014-11-01

    Glaucoma is an acquired progressive optic neuropathy which is characterized by changes in the optic nerve head and retinal nerve fiber layer (RNFL). White-on-white perimetry is the gold standard for the diagnosis of glaucoma. However, it can detect defects in the visual field only after the loss of as many as 40% of the ganglion cells. Hence, the measurement of RNFL thickness has come up. Optical coherence tomography and scanning laser polarimetry (SLP) are the techniques that utilize the evaluation of RNFL for the evaluation of glaucoma. SLP provides RNFL thickness measurements based upon the birefringence of the retinal ganglion cell axons. We have reviewed the published literature on the use of SLP in glaucoma. This review elucidates the technological principles, recent developments and the role of SLP in the diagnosis and monitoring of glaucomatous optic neuropathy, in the light of scientific evidence so far.

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

    Science.gov (United States)

    de Jonge, Niels [Oak Ridge, TN

    2010-08-17

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

  7. Spatiotemporal closure of fractional laser-ablated channels imaged by optical coherence tomography and reflectance confocal microscopy

    DEFF Research Database (Denmark)

    Banzhaf, Christina A.; Wind, Bas S.; Mogensen, Mette

    2016-01-01

    Background and Objective Optical coherence tomography (OCT) and reflectance confocal microscopy (RCM) offer high-resolution optical imaging of the skin, which may provide benefit in the context of laser-assisted drug delivery. We aimed to characterize postoperative healing of ablative fractional...... laser (AFXL)-induced channels and dynamics in their spatiotemporal closure using in vivo OCT and RCM techniques. Study design/Materials and Methods The inner forearm of healthy subjects (n = 6) was exposed to 10,600 nm fractional CO2 laser using 5 and 25% densities, 120 μm beam diameter, 5, 15, and 25 m......J/microbeam. Treatment sites were scanned with OCT to evaluate closure of AFXL-channels and RCM to evaluate subsequent re-epithelialization. Results OCT and RCM identified laser channels in epidermis and upper dermis as black, ablated tissue defects surrounded by characteristic hyper-and hyporeflective zones. OCT imaged...

  8. Dark-field scanning confocal microscope for vertical particle tracks in nuclear emulsion

    International Nuclear Information System (INIS)

    Astakhov, A.Ya.; Batusov, Yu.A.; Soroko, L.M.; Tereshchenko, S.V.; Tereshchenko, V.V.

    1999-01-01

    The principle of the DArk-FIeld Scanning CONfocal (DAFISCON) microscope for selective observation of the vertical particle tracks in nuclear emulsion is described. The construction of the DAFISCON microscope, built on the basis of the 2D measurement microscope, is described. The results of the experimental testing of the DAFISCON microscope, accomplished at high density of the vertical particle tracks, are presented. The 2D plot and the 1D plot of the CCD dark-field image are given. The spatial resolution of our microscope can be increased by using the objective with higher aperture

  9. A confocal laser scanning microscopic study on thermoresponsive

    Indian Academy of Sciences (India)

    Monodisperse poly(N -isopropylacrylamide) (PNIPAM) particles loaded with cadmium telluride (CdTe) quantum dots (QDs) of two different sizes (4.7 nm and 5.6 nm) were synthesized in aqueous medium by bonding the capping agent on the quantum dots to the amide groups of PNIPAM and incubating the samples at 45° ...

  10. Laser scanning of experimental solar cells

    Science.gov (United States)

    Plunkett, B. C.; Lasswell, P. G.

    1980-01-01

    A description is presented of a laser scanning instrument which makes it possible to display and measure the spatial response of a solar cell. Examples are presented to illustrate the use of generated micrographs in the isolation of flaws and features of the cell. The laser scanner system uses a 4 mW, CW helium-neon laser, operating a wavelength of 0.633 micrometers. The beam is deflected by two mirror galvanometers arranged to scan in orthogonal directions. After being focused on the solar cell by the beam focusing lens, the moving light spot raster scans the specimen. The current output of the photovoltaic device under test, as a function of the scan dot position, can be displayed in several modes. The laser scanner has proved to be a very useful diagnostic tool in optimizing the process design of transparent metal film photovoltaic devices on Zn3P2, a relatively new photovoltaic material.

  11. Multicolor Scanning Laser Imaging in Diabetic Retinopathy.

    Science.gov (United States)

    Ahmad, Mohammad S Z; Carrim, Zia Iqbal

    2017-11-01

    Diabetic retinopathy is a common cause of blindness in individuals younger than 60 years. Screening for retinopathy is undertaken using conventional color fundus photography and relies on the identification of hemorrhages, vascular abnormalities, exudates, and cotton-wool spots. These can sometimes be difficult to identify. Multicolor scanning laser imaging, a new imaging modality, may have a role in improving screening outcomes, as well as facilitating treatment decisions. Observational case series comprising two patients with known diabetes who were referred for further examination after color fundus photography revealed abnormal findings. Multicolor scanning laser imaging was undertaken. Features of retinal disease from each modality were compared. Multicolor scanning laser imaging provides superior visualization of retinal anatomy and pathology, thereby facilitating risk stratification and treatment decisions. Multicolor scanning laser imaging is a novel imaging technique offering the potential for improving the reliability of screening for diabetic retinopathy. Validation studies are warranted.

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

    Science.gov (United States)

    Wang, Peng; Behan, Gavin; Kirkland, Angus I; Nellist, Peter D; Cosgriff, Eireann C; D'Alfonso, Adrian J; Morgan, Andrew J; Allen, Leslie J; Hashimoto, Ayako; Takeguchi, Masaki; Mitsuishi, Kazutaka; Shimojo, Masayuki

    2011-06-01

    Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored. Copyright © 2010 Elsevier B.V. All rights reserved.

  13. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy.

    Science.gov (United States)

    Schulz, Olaf; Pieper, Christoph; Clever, Michaela; Pfaff, Janine; Ruhlandt, Aike; Kehlenbach, Ralph H; Wouters, Fred S; Großhans, Jörg; Bunt, Gertrude; Enderlein, Jörg

    2013-12-24

    We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.

  14. Laser scanning camera inspects hazardous area

    International Nuclear Information System (INIS)

    Fryatt, A.; Miprode, C.

    1985-01-01

    Main operational characteristics of a new laser scanning camera are presented. The camera is intended primarily for low level high resolution viewing inside nuclear reactors. It uses a He-Ne laser beam raster; by detecting the reflected light by means of a phomultiplier, the subject under observation can be reconstructed in an electronic video store and reviewed on a conventional monitor screen

  15. In vivo laser confocal microscopy findings of a cornea with osteogenesis imperfecta

    Directory of Open Access Journals (Sweden)

    Kobayashi A

    2014-02-01

    Full Text Available Akira Kobayashi, Tomomi Higashide, Hideaki Yokogawa, Natsuko Yamazaki, Toshinori Masaki, Kazuhisa Sugiyama Department of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan Objective: To report the in vivo laser confocal microscopy findings of a cornea with osteogenesis imperfecta (OI with special attention to the abnormality of Bowman's layer and sub-Bowman's fibrous structures (K-structures. Patients and methods: Two patients (67-year-old male and his 26-year-old son with OI type I were included in this study. Slit lamp biomicroscopic and in vivo laser confocal microscopic examinations were performed for both patients. Central corneal thickness and central endothelial cell density were also measured. Results: Although the corneas looked clear with normal endothelial density for both eyes in both patients, they were quite thin (386 µm oculus dexter (OD (the right eye and 384 µm oculus sinister (OS (the left eye in the father and 430 µm OD and 425 µm OS in the son. In both patients, slit lamp biomicroscopic and in vivo laser confocal microscopic examination showed similar results. Anterior corneal mosaics produced by rubbing the eyelid under fluorescein were completely absent in both eyes. In vivo laser confocal microscopy revealed an absent or atrophic Bowman's layer; a trace of a presumed Bowman's layer and/or basement membrane was barely visible with high intensity. Additionally, K-structures were completely absent in both eyes. Conclusion: The absence of K-structures and fluorescein anterior corneal mosaics strongly suggested an abnormality of Bowman's layer in these OI patients. Keywords: osteogenesis imperfecta, K-structure, confocal microscopy, Bowman's layer

  16. Scanning laser microscope for imaging nanostructured superconductors

    International Nuclear Information System (INIS)

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

    2010-01-01

    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.

  17. Scanning laser microscope for imaging nanostructured superconductors

    Science.gov (United States)

    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.

  18. RADIOMETRIC CALIBRATION OF AIRBORNE LASER SCANNING DATA

    OpenAIRE

    Pilarska Magdalena

    2016-01-01

    Airborne laser scanning (ALS) is widely used passive remote sensing technique. The radiometric calibration of ALS data is presented in this article. This process is a necessary element in data processing since it eliminates the influence of the external factors on the obtained values of radiometric features such as range and incidence angle. The datasets were captured with three different laser scanners; since each of these operates at a different wavelength (532, 106 4 and 1550 nm) th...

  19. Noninvasive imaging of the human rod photoreceptor mosaic using a confocal adaptive optics scanning ophthalmoscope

    Science.gov (United States)

    Dubra, Alfredo; Sulai, Yusufu; Norris, Jennifer L.; Cooper, Robert F.; Dubis, Adam M.; Williams, David R.; Carroll, Joseph

    2011-01-01

    The rod photoreceptors are implicated in a number of devastating retinal diseases. However, routine imaging of these cells has remained elusive, even with the advent of adaptive optics imaging. Here, we present the first in vivo images of the contiguous rod photoreceptor mosaic in nine healthy human subjects. The images were collected with three different confocal adaptive optics scanning ophthalmoscopes at two different institutions, using 680 and 775 nm superluminescent diodes for illumination. Estimates of photoreceptor density and rod:cone ratios in the 5°–15° retinal eccentricity range are consistent with histological findings, confirming our ability to resolve the rod mosaic by averaging multiple registered images, without the need for additional image processing. In one subject, we were able to identify the emergence of the first rods at approximately 190 μm from the foveal center, in agreement with previous histological studies. The rod and cone photoreceptor mosaics appear in focus at different retinal depths, with the rod mosaic best focus (i.e., brightest and sharpest) being at least 10 μm shallower than the cones at retinal eccentricities larger than 8°. This study represents an important step in bringing high-resolution imaging to bear on the study of rod disorders. PMID:21750765

  20. Scanning confocal slit photon counter measurements of post-PRK haze in two-year study

    Science.gov (United States)

    Taboada, John; Gaines, David; Perez, Mary A.; Waller, Steve G.; Ivan, Douglas J.; Baldwin, J. Bruce; LoRusso, Frank; Tutt, Ronald C.; Thompson, B.; Perez, Jose; Tredici, Thomas; Johnson, Dan A.

    2001-06-01

    In our study, a group of 80 United States Air Force, non- flying personnel will undergo photorefractive corneal surgery for moderate levels of myopia (< 6 diopters) and 20 will serve as controls. As of this report, approximately 56 have had the treatment. Of these, only about 59% of the treated eyes showed even a trace (.5) level of clinically assessed haze at any time. We report on the use of a recently developed instrument designed for the objective measurement of these low levels of haze in treated corneas. The sensitivity of the instrument is derived from the use of a scanning confocal slit photon counter. The use of a physical standard for calibration secures accuracy and reproducibility over an extensive period of time. Our haze measurements in this study revealed a very low level increase from baseline values for these patients. The typical increase over baseline was of the same magnitude as the variability in the observations, although the inherent variability in the measurements was approximately 0.25 times the value of the patient's haze variability.

  1. High-speed adaptive optics line scan confocal retinal imaging for human eye.

    Science.gov (United States)

    Lu, Jing; Gu, Boyu; Wang, Xiaolin; Zhang, Yuhua

    2017-01-01

    Continuous and rapid eye movement causes significant intraframe distortion in adaptive optics high resolution retinal imaging. To minimize this artifact, we developed a high speed adaptive optics line scan confocal retinal imaging system. A high speed line camera was employed to acquire retinal image and custom adaptive optics was developed to compensate the wave aberration of the human eye's optics. The spatial resolution and signal to noise ratio were assessed in model eye and in living human eye. The improvement of imaging fidelity was estimated by reduction of intra-frame distortion of retinal images acquired in the living human eyes with frame rates at 30 frames/second (FPS), 100 FPS, and 200 FPS. The device produced retinal image with cellular level resolution at 200 FPS with a digitization of 512×512 pixels/frame in the living human eye. Cone photoreceptors in the central fovea and rod photoreceptors near the fovea were resolved in three human subjects in normal chorioretinal health. Compared with retinal images acquired at 30 FPS, the intra-frame distortion in images taken at 200 FPS was reduced by 50.9% to 79.7%. We demonstrated the feasibility of acquiring high resolution retinal images in the living human eye at a speed that minimizes retinal motion artifact. This device may facilitate research involving subjects with nystagmus or unsteady fixation due to central vision loss.

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

    Science.gov (United States)

    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.

  3. Line-scanning confocal microscopy for high-resolution imaging of upconverting rare-earth-based contrast agents

    Science.gov (United States)

    Higgins, Laura M.; Zevon, Margot; Ganapathy, Vidya; Sheng, Yang; Tan, Mei Chee; Riman, Richard E.; Roth, Charles M.; Moghe, Prabhas V.; Pierce, Mark C.

    2015-01-01

    Abstract. Rare-earth (RE) doped nanocomposites emit visible luminescence when illuminated with continuous wave near-infrared light, making them appealing candidates for use as contrast agents in biomedical imaging. However, the emission lifetime of these materials is much longer than the pixel dwell times used in scanning intravital microscopy. To overcome this limitation, we have developed a line-scanning confocal microscope for high-resolution, optically sectioned imaging of samples labeled with RE-based nanomaterials. Instrument performance is quantified using calibrated test objects. NaYF4:Er,Yb nanocomposites are imaged in vitro, and in ex vivo tissue specimens, with direct comparison to point-scanning confocal microscopy. We demonstrate that the extended pixel dwell time of line-scanning confocal microscopy enables subcellular-level imaging of these nanomaterials while maintaining optical sectioning. The line-scanning approach thus enables microscopic imaging of this emerging class of contrast agents for preclinical studies, with the potential to be adapted for real-time in vivo imaging in the clinic. PMID:26603495

  4. Multiplatform Mobile Laser Scanning: Usability and Performance

    Directory of Open Access Journals (Sweden)

    Yuwei Chen

    2012-08-01

    Full Text Available Mobile laser scanning is an emerging technology capable of capturing three-dimensional data from surrounding objects. With state-of-the-art sensors, the achieved point clouds capture object details with good accuracy and precision. Many of the applications involve civil engineering in urban areas, as well as traffic and other urban planning, all of which serve to make 3D city modeling probably the fastest growing market segment in this field. This article outlines multiplatform mobile laser scanning solutions such as vehicle- and trolley-operated urban area data acquisition, and boat-mounted equipment for fluvial environments. Moreover, we introduce a novel backpack version of mobile laser scanning equipment for surveying applications in the field of natural sciences where the requirements include precision and mobility in variable terrain conditions. In addition to presenting a technical description of the systems, we discuss the performance of the solutions in the light of various applications in the fields of urban mapping and modeling, fluvial geomorphology, snow-cover characterization, precision agriculture, and in monitoring the effects of climate change on permafrost landforms. The data performance of the mobile laser scanning approach is described by the results of an evaluation of the ROAMER on a permanent MLS test field. Furthermore, an in situ accuracy assessment using a field of spherical 3D targets for the newly-introduced Akhka backpack system is conducted and reported on.

  5. Polarized differential-phase laser scanning microscope

    International Nuclear Information System (INIS)

    Chou Chien; Lyu, C.-W.; Peng, L.-C.

    2001-01-01

    A polarized differential-phase laser scanning microscope, which combines a polarized optical heterodyne Mach-Zehnder interferometer and a differential amplifier to scan the topographic image of a surface, is proposed. In the experiment the differential amplifier, which acts as a PM-AM converter, in the experiment, converting phase modulation (PM) into amplitude modulation (AM). Then a novel, to our knowledge, phase demodulator was proposed and implemented for the differential-phase laser scanning microscope. An optical grating (1800 lp/mm) was imaged. The lateral and the depth resolutions of the imaging system were 0.5 μm and 1 nm, respectively. The detection accuracy, which was limited by the reflectivity variation of the test surface, is discussed

  6. Correlative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxide

    KAUST Repository

    Rodighiero, Simona

    2015-03-22

    Confocal microscopy imaging of cells allows to visualize the presence of specific antigens by using fluorescent tags or fluorescent proteins, with resolution of few hundreds of nanometers, providing their localization in a large field-of-view and the understanding of their cellular function. Conversely, in scanning electron microscopy (SEM), the surface morphology of cells is imaged down to nanometer scale using secondary electrons. Combining both imaging techniques have brought to the correlative light and electron microscopy, contributing to investigate the existing relationships between biological surface structures and functions. Furthermore, in SEM, backscattered electrons (BSE) can image local compositional differences, like those due to nanosized gold particles labeling cellular surface antigens. To perform SEM imaging of cells, they could be grown on conducting substrates, but obtaining images of limited quality. Alternatively, they could be rendered electrically conductive, coating them with a thin metal layer. However, when BSE are collected to detect gold-labeled surface antigens, heavy metals cannot be used as coating material, as they would mask the BSE signal produced by the markers. Cell surface could be then coated with a thin layer of chromium, but this results in a loss of conductivity due to the fast chromium oxidation, if the samples come in contact with air. In order to overcome these major limitations, a thin layer of indium-tin-oxide was deposited by ion-sputtering on gold-decorated HeLa cells and neurons. Indium-tin-oxide was able to provide stable electrical conductivity and preservation of the BSE signal coming from the gold-conjugated markers. © 2015 Wiley Periodicals, Inc.

  7. Laser-induced cartilage damage: an ex-vivo model using confocal microscopy

    Science.gov (United States)

    Frenz, Martin; Zueger, Benno J.; Monin, D.; Weiler, C.; Mainil-Varlet, P. M.; Weber, Heinz P.; Schaffner, Thomas

    1999-06-01

    Although there is an increasing popularity of lasers in orthopedic surgery, there is a growing concern about negative side effects of this therapy e.g. prolonged restitution time, radiation damage to adjacent cartilage or depth effects like bone necrosis. Despite case reports and experimental investigations over the last few years little is known about the extent of acute cartilage damage induced by different lasers types and energies. Histological examination offers only limited insights in cell viability and metabolism. Ho:YAG and Er:YAG lasers emitting at 2.1 micrometer and 2.94 micrometer, respectively, are ideally suited for tissue treatment because these wavelengths are strongly absorbed in water. The Purpose of the present study is to evaluate the effect of laser type and energy on chondrocyte viability in an ex vivo model. Free running Er:YAG (E equals 100 and 150 mJ) and Ho:YAG (E equals 500 and 800 mJ) lasers were used at different energy levels using a fixed pulse length of 400 microseconds. The energy was delivered at 8 Hz through optical fibers. Fresh bovine hyaline cartilage samples were mounted in a water bath at room temperature and the fiber was positioned at 30 degree and 180 degree angles relative to the tissue surface. After laser irradiation the samples were assessed by a life-dead cell viability test using a confocal microscope and by standard histology. Thermal damage was much deeper with Ho:YAG (up to 1800 micrometer) than with the Er:YAG laser (up to 70 micrometer). The cell viability test revealed a damage zone about twice the one determined by standard histology. Confocal microscopy is a powerful tool for assessing changes in tissue structure after laser treatment. In addition this technique allows to quantify these alterations without necessitating time consuming and expensive animal experiments.

  8. Nondestructive 3D confocal laser imaging with deconvolution of seven whole stardust tracks with complementary XRF and quantitative analysis

    International Nuclear Information System (INIS)

    Greenberg, M.; Ebel, D.S.

    2009-01-01

    We present a nondestructive 3D system for analysis of whole Stardust tracks, using a combination of Laser Confocal Scanning Microscopy and synchrotron XRF. 3D deconvolution is used for optical corrections, and results of quantitative analyses of several tracks are presented. The Stardust mission to comet Wild 2 trapped many cometary and ISM particles in aerogel, leaving behind 'tracks' of melted silica aerogel on both sides of the collector. Collected particles and their tracks range in size from submicron to millimeter scale. Interstellar dust collected on the obverse of the aerogel collector is thought to have an average track length of ∼15 (micro)m. It has been our goal to perform a total non-destructive 3D textural and XRF chemical analysis on both types of tracks. To that end, we use a combination of Laser Confocal Scanning Microscopy (LCSM) and X Ray Florescence (XRF) spectrometry. Utilized properly, the combination of 3D optical data and chemical data provides total nondestructive characterization of full tracks, prior to flattening or other destructive analysis methods. Our LCSM techniques allow imaging at 0.075 (micro)m/pixel, without the use of oil-based lenses. A full textural analysis on track No.82 is presented here as well as analysis of 6 additional tracks contained within 3 keystones (No.128, No.129 and No.140). We present a method of removing the axial distortion inherent in LCSM images, by means of a computational 3D Deconvolution algorithm, and present some preliminary experiments with computed point spread functions. The combination of 3D LCSM data and XRF data provides invaluable information, while preserving the integrity of the samples for further analysis. It is imperative that these samples, the first extraterrestrial solids returned since the Apollo era, be fully mapped nondestructively in 3D, to preserve the maximum amount of information prior to other, destructive analysis.

  9. Scanning laser ophthalmoscope design with adaptive optics

    OpenAIRE

    Laut, SP; Jones, SM; Olivier, SS; Werner, JS

    2005-01-01

    A design for a high-resolution scanning instrument is presented for in vivo imaging of the human eye at the cellular scale. This system combines adaptive optics technology with a scanning laser ophthalmoscope (SLO) to image structures with high lateral (∼2 μm) resolution. In this system, the ocular wavefront aberrations that reduce the resolution of conventional SLOs are detected by a Hartmann-Shack wavefront sensor, and compensated with two deformable mirrors in a closed-loop for dynamic cor...

  10. Confocal Laser Endomicroscopy in the Study of Colonic Mucosa in IBD Patients: A Review

    Directory of Open Access Journals (Sweden)

    Francesca Salvatori

    2012-01-01

    Full Text Available Confocal laser endomicroscopy (CLE is one of several novel methods that provide real-time, high-resolution imaging at a micronscale via endoscopes. CLE and related technologies are often termed “virtual biopsy” as they simulate the images seen in traditional histology. Recently, the use of CLE was reported in the study of colonic mucosa in patients with inflammatory bowel diseases and in particular in patients affected by ulcerative colitis. CLE has the potential to have an important role in management of IBD patients as it can be used to assess the grading of colitis and in detection of microscopic colitis in endoscopically silent segments. Moreover, CLE can be used in surveillance programs especially in high-risk patients. This report aims to evaluate the current data on the application of confocal endomicroscopy in clinical gastroenterology and particularly in the study of colonic mucosa in UC patients.

  11. Efficiency of the confocal method of laser endomicroscopy in complex diagnoses of diseases of common bile duct

    International Nuclear Information System (INIS)

    Anaskin, S G; Korniletsky, I D; Panchenkov, D N; Chertyuk, V B; Sazonov, D V; Zabozlayev, F G; Danilevskaya, O V; Mokshina, N V

    2017-01-01

    One of the more frequent manifestations of diseases of the bile ducts are its’ strictures or stenoses that could be of either malignant or benign nature. Current methods of diagnosing this pathology include computer tomography (CT) scan, magnetic resonance cholangiopancreatography (MRCP), endoscopic ultrasound (EUS) and endoscopic retrograde cholangiopancreatography (ERCP). However, these methods are not always informative, which makes this a current and topical problem. A fundamentally new method that broadens the capabilities of ERCP when diagnosing diseases of the bile duct accompanied by the development of strictures or stenoses is probe-based confocal laser endomicroscopy (pCLE). The method is based on the principle of confocal fluorescence microscopy. The most elaborate complications arise with the presence of the pre-existing pancreatobiliary pathology: pseudotumoral chronic pancreatitis, acute cholangitis, etc. Early stage cholangiocarcinoma diagnosis can be difficult (and not always possible) even with the help of modern research methods. For the timely diagnostic it is advantageous to conduct pCLE and targeted biopsy of the zone with most manifested changes. In all instances, the first use of the pCLE method for diagnostic purposes allowed us to clarify and correctly verify the diagnosis. When concerning the diseases of the bile duct, the modern stage of pCLE development can be of critical importance when other methods are not effective. (paper)

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

    Science.gov (United States)

    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.

  13. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: SPECTROSCOPY

    Science.gov (United States)

    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...

  14. Observation of ice sheet formation on methane and ethane gas hydrates using a scanning confocal microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Nagao, J.; Shimomura, N.; Ebinuma, T.; Narita, H. [National Inst. of Advanced Industrial Science and Technology, Toyohira, Sapporo (Japan). Methane Hydrate Research Lab.

    2008-07-01

    Interest in gas hydrates has increased in recent years due to the discovery of large deposits under the ocean floor and in permafrost regions. Natural gas hydrates, including methane, is expected to become a new energy source and a medium for energy storage and transportation. Gas hydrates consist of an open network of water molecules that are hydrogen-bonded in a similar manner to ice. Gas molecules are interstitially engaged under high pressures and low temperatures. Although the dissociation temperature of methane hydrate under atmospheric pressure is about 193 K, studies have shown that methane hydrate can be stored at atmospheric pressure and 267 K for 2 years. Because of this phenomenon, known as self-preservation, transportation and storage of methane hydrate can occur at temperature conditions milder than those for liquefied methane gas at atmospheric pressure. This study examined the surface changes of methane and ethane hydrates during dissociation using an optical microscope and confocal scanning microscope (CSM). This paper reported on the results when the atmospheric gas pressure was decreased. Ice sheets formed on the surfaces of methane and ethane gas hydrates due to depressurizing dissociation of methane and ethane hydrates when the methane and ethane gas pressures were decreased at designated temperatures. The dissociation of methane gas hydrate below below 237 K resulted in the generation of small ice particles on the hydrate surface. A transparent ice sheet formed on the hydrate surface above 242 K. The thickness of the ice sheet on the methane hydrate surface showed the maximum of ca. 30 {mu}m at 253 K. In the case of ethane hydrates, ice particles and ice sheets formed below 262 and 267 respectively. Since the ice particles and ice sheets were formed by water molecules generated during the gas hydrate dissociation, the mechanism of ice sheet formation depends on the dissociation rate of hydrate, ice particle sintering rate, and water molecule

  15. Morphological characteristics of the optic nerve evaluated by confocal laser tomography (HRT3) and laser polarimetry (GDx-VCC) in a normal population from the city of Barcelona.

    Science.gov (United States)

    Fallon, M; Pazos, M; Morilla, A; Sebastián, M A; Xancó, R; Mora, C; Calderón, B; Vega, Z; Antón, A

    2015-11-01

    To evaluate morphological parameters of optic disc and retinal nerve fiber layer (RNFL) examined with confocal laser tomography (HRT3) and laser polarimetry (GDx-VCC) in a normal population, and analyze correlations of these parameters with demographic variables. Cross-sectional study in the context of a glaucoma screening campaign in the primary care center of Barcelona. The individuals selected were non-hypertensive Mediterranean Caucasians with risk for glaucoma development (individuals≥60 years old or≥40 years old with family history of glaucoma or intraocular pressure or myopia>3diopter). All subjects underwent a complete ophthalmic examination, confocal laser tomography (HRT3) and scanning laser polarimetry (GDX-VCC), subjects with results within normal limits only being included. Structural parameters were analyzed along with age, refraction, and pachymetry based on the Spearman rank correlation test. A total of 224 subjects included, with a mean age of 63.4±11.1 years. Disc areas, excavation and ring area were 2.14±0.52mm(2), 0.44±0.34mm (2) and 1.69±0.38mm(2), respectively. The mean RNFL (GDX) was 55.9±6.9μm. Age was correlated with lower ring volume, highest rate of cup shape measure, largest mean and maximum cup depth, lower nerve fiber index (NFI) and RNFL (all p-values below .05). The mean values and distribution of several parameters of the papilla and the RNFL in normal Mediterranean Caucasians population are presented. A loss of thickness of the RNFL, ring thinning, and enlarged cup was observed with increased age. Copyright © 2014 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.

  16. Role of Confocal Laser Endomicroscopy in Detection of Residual Barrett's Esophagus after Radiofrequency Ablation

    Directory of Open Access Journals (Sweden)

    Giorgio Diamantis

    2011-01-01

    Full Text Available Endoscopic endoluminal radiofrequency ablation (RFA is a novel and promising modality for Barrett's esophagus (BE treatment. Actually the only surveillance method after the ablation treatment is random biopsies throughout the whole treated area. Confocal laser endomicroscopy (CLE is a new endoscopic imaging tool that permits high-resolution microscopic examination of the gastrointestinal tract. The technology has garnered increasing attention because of its ability to provide real-time “optical” biopsy specimens, with a very high sensitivity and specificity. This paper summarize the potential application of CLE in the surveillance of the reepithelialization of BE, after endoscopic RFA.

  17. Confocal Laser Endomicroscopy in Neurosurgery: A New Technique with Much Potential

    Directory of Open Access Journals (Sweden)

    David Breuskin

    2013-01-01

    Full Text Available Technical innovations in brain tumour diagnostic and therapy have led to significant improvements of patient outcome and recurrence free interval. The use of technical devices such as surgical microscopes as well as neuronavigational systems have helped localising tumours as much as fluorescent agents, such as 5-aminolaevulinic acid, have helped visualizing pathologically altered tissue. Nonetheless, intraoperative instantaneous frozen sections and histological diagnosis remain the only method of gaining certainty of the nature of the resected tissue. This technique is time consuming and does not provide close-to-real-time information. In gastroenterology, confocal endoscopy closed the gap between tissue resection and histological examination, providing an almost real-time histological diagnosis. The potential of this technique using a confocal laser endoscope EndoMAG1 by Karl Storz Company was evaluated by our group on pig brains, tumour tissue cell cultures, and fresh human tumour specimen. Here, the authors report for the first time on the results of applying this new technique and provide first confocal endoscopic images of various brain and tumour structures. In all, the technique harbours a very promising potential to provide almost real-time intraoperative diagnosis, but further studies are needed to provide evidence for the technique’s potential.

  18. Laser confocal measurement system for curvature radius of lenses based on grating ruler

    Science.gov (United States)

    Tian, Jiwei; Wang, Yun; Zhou, Nan; Zhao, Weirui; Zhao, Weiqian

    2015-02-01

    In the modern optical measurement field, the radius of curvature (ROC) is one of the fundamental parameters of optical lens. Its measurement accuracy directly affects the other optical parameters, such as focal length, aberration and so on, which significantly affect the overall performance of the optical system. To meet the demand of measurement instruments for radius of curvature (ROC) with high accuracy in the market, we develop a laser confocal radius measurement system with grating ruler. The system uses the peak point of the confocal intensity curve to precisely identify the cat-eye and confocal positions and then measure the distance between these two positions by using the grating ruler, thereby achieving the high-precision measurement for the ROC. The system has advantages of high focusing sensitivity and anti-environment disturbance ability. And the preliminary theoretical analysis and experiments show that the measuring repeatability can be up to 0.8 um, which can provide an effective way for the accurate measurement of ROC.

  19. Development of confocal laser microscope system for examination of microscopic characteristics of radiophotoluminescence glass dosemeters

    International Nuclear Information System (INIS)

    Maki, D.; Ishii, T.; Sato, F.; Kato, Y.; Yamamoto, T.; Iida, T.

    2011-01-01

    A confocal laser microscope system was developed for the measurement of radiophotoluminescence (RPL) photons emitted from a minute alpha-ray-irradiated area in an RPL glass dosemeter. The system was composed mainly of an inverted-type microscope, an ultraviolet laser, an XY movable stage and photon-counting circuits. The photon-counting circuits were effective in the reduction of the background noise level in the measurement of RPL photons. The performance of this microscope system was examined by the observation of standard RPL glass samples irradiated using 241 Am alpha rays. The spatial resolution of this system was ∼3 μm, and with regard to the sensitivity of this system, a hit of more than four to five alpha rays in unit area produced enough amount of RPL photons to construct the image. (authors)

  20. Development of confocal laser microscope system for examination of microscopic characteristics of radiophotoluminescence glass dosemeters.

    Science.gov (United States)

    Maki, Daisuke; Ishii, Tetsuya; Sato, Fuminobu; Kato, Yushi; Yamamoto, Takayoshi; Iida, Toshiyuki

    2011-03-01

    A confocal laser microscope system was developed for the measurement of radiophotoluminescence (RPL) photons emitted from a minute alpha-ray-irradiated area in an RPL glass dosemeter. The system was composed mainly of an inverted-type microscope, an ultraviolet laser, an XY movable stage and photon-counting circuits. The photon-counting circuits were effective in the reduction of the background noise level in the measurement of RPL photons. The performance of this microscope system was examined by the observation of standard RPL glass samples irradiated using (241)Am alpha rays. The spatial resolution of this system was ∼ 3 μm, and with regard to the sensitivity of this system, a hit of more than four to five alpha rays in unit area produced enough amount of RPL photons to construct the image.

  1. Nanoscale Energy-Filtered Scanning Confocal Electron Microscopy Using a Double-Aberration-Corrected Transmission Electron Microscope

    International Nuclear Information System (INIS)

    Wang Peng; Behan, Gavin; Kirkland, Angus I.; Nellist, Peter D.; Takeguchi, Masaki; Hashimoto, Ayako; Mitsuishi, Kazutaka; Shimojo, Masayuki

    2010-01-01

    We demonstrate that a transmission electron microscope fitted with two spherical-aberration correctors can be operated as an energy-filtered scanning confocal electron microscope. A method for establishing this mode is described and initial results showing 3D chemical mapping with nanoscale sensitivity to height and thickness changes in a carbon film are presented. Importantly, uncorrected chromatic aberration does not limit the depth resolution of this technique and moreover performs an energy-filtering role, which is explained in terms of a combined depth and energy-loss response function.

  2. Mobile Laser Scanning for Indoor Modelling

    Directory of Open Access Journals (Sweden)

    C. Thomson

    2013-10-01

    Full Text Available The process of capturing and modelling buildings has gained increased focus in recent years with the rise of Building Information Modelling (BIM. At the heart of BIM is a process change for the construction and facilities management industries whereby a BIM aids more collaborative working through better information exchange, and as a part of the process Geomatic/Land Surveyors are not immune from the changes. Terrestrial laser scanning has been proscribed as the preferred method for rapidly capturing buildings for BIM geometry. This is a process change from a traditional measured building survey just with a total station and is aided by the increasing acceptance of point cloud data being integrated with parametric building models in BIM tools such as Autodesk Revit or Bentley Architecture. Pilot projects carried out previously by the authors to investigate the geometry capture and modelling of BIM confirmed the view of others that the process of data capture with static laser scan setups is slow and very involved requiring at least two people for efficiency. Indoor Mobile Mapping Systems (IMMS present a possible solution to these issues especially in time saved. Therefore this paper investigates their application as a capture device for BIM geometry creation over traditional static methods through a fit-for-purpose test.

  3. Mobile Laser Scanning for Indoor Modelling

    Science.gov (United States)

    Thomson, C.; Apostolopoulos, G.; Backes, D.; Boehm, J.

    2013-10-01

    The process of capturing and modelling buildings has gained increased focus in recent years with the rise of Building Information Modelling (BIM). At the heart of BIM is a process change for the construction and facilities management industries whereby a BIM aids more collaborative working through better information exchange, and as a part of the process Geomatic/Land Surveyors are not immune from the changes. Terrestrial laser scanning has been proscribed as the preferred method for rapidly capturing buildings for BIM geometry. This is a process change from a traditional measured building survey just with a total station and is aided by the increasing acceptance of point cloud data being integrated with parametric building models in BIM tools such as Autodesk Revit or Bentley Architecture. Pilot projects carried out previously by the authors to investigate the geometry capture and modelling of BIM confirmed the view of others that the process of data capture with static laser scan setups is slow and very involved requiring at least two people for efficiency. Indoor Mobile Mapping Systems (IMMS) present a possible solution to these issues especially in time saved. Therefore this paper investigates their application as a capture device for BIM geometry creation over traditional static methods through a fit-for-purpose test.

  4. Miniature in vivo MEMS-based line-scanned dual-axis confocal microscope for point-of-care pathology

    Science.gov (United States)

    Yin, C.; Glaser, A.K.; Leigh, S. Y.; Chen, Y.; Wei, L.; Pillai, P. C. S.; Rosenberg, M. C.; Abeytunge, S.; Peterson, G.; Glazowski, C.; Sanai, N.; Mandella, M. J.; Rajadhyaksha, M.; Liu, J. T. C.

    2016-01-01

    There is a need for miniature optical-sectioning microscopes to enable in vivo interrogation of tissues as a real-time and noninvasive alternative to gold-standard histopathology. Such devices could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Miniature confocal microscopes have been developed by various researchers and corporations to enable optical sectioning of highly scattering tissues, all of which have necessitated various trade-offs in size, speed, depth selectivity, field of view, resolution, image contrast, and sensitivity. In this study, a miniature line-scanned (LS) dual-axis confocal (DAC) microscope, with a 12-mm diameter distal tip, has been developed for clinical point-of-care pathology. The dual-axis architecture has demonstrated an advantage over the conventional single-axis confocal configuration for reducing background noise from out-of-focus and multiply scattered light. The use of line scanning enables fast frame rates (16 frames/sec is demonstrated here, but faster rates are possible), which mitigates motion artifacts of a hand-held device during clinical use. We have developed a method to actively align the illumination and collection beams in a DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo clinical use, enables our device to achieve an optical-sectioning thickness and lateral resolution of 2.0 and 1.1 microns respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate the clinical potential of this high-speed optical-sectioning microscopy device. PMID:26977337

  5. Molecular confocal laser endomicroscopy: a novel technique for in vivo cellular characterization of gastrointestinal lesions.

    Science.gov (United States)

    Karstensen, John Gásdal; Klausen, Pia Helene; Saftoiu, Adrian; Vilmann, Peter

    2014-06-28

    While flexible endoscopy is essential for macroscopic evaluation, confocal laser endomicroscopy (CLE) has recently emerged as an endoscopic method enabling visualization at a cellular level. Two systems are currently available, one based on miniprobes that can be inserted via a conventional endoscope or via a needle guided by endoscopic ultrasound. The second system has a confocal microscope integrated into the distal part of an endoscope. By adding molecular probes like fluorescein conjugated antibodies or fluorescent peptides to this procedure (either topically or systemically administered during on-going endoscopy), a novel world of molecular evaluation opens up. The method of molecular CLE could potentially be used for estimating the expression of important receptors in carcinomas, subsequently resulting in immediate individualization of treatment regimens, but also for improving the diagnostic accuracy of endoscopic procedures by identifying otherwise invisible mucosal lesions. Furthermore, studies have shown that fluorescein labelled drugs can be used to estimate the affinity of the drug to a target organ, which probably can be correlated to the efficacy of the drug. However, several of the studies in this research field have been conducted in animal facilities or in vitro, while only a limited number of trials have actually been carried out in vivo. Therefore, safety issues still needs further evaluations. This review will present an overview of the implications and pitfalls, as well as future challenges of molecular CLE in gastrointestinal diseases.

  6. Experimental verification of subthreshold laser therapy using conventional pattern scan laser.

    Directory of Open Access Journals (Sweden)

    Tomoyasu Shiraya

    Full Text Available Leading-edge therapeutic laser technologies are not available at every medical facility; therefore, alternative approaches incorporating novel advances in digital and laser technology into more readily available conventional methods have generated significant research interest. Using a rabbit model, this study investigated whether the algorithm used in the Endpoint Management (EM software system of the latest devices could enable subthreshold laser treatment in conventional retinal tissue laser therapy systems.Two types of devices were used, the PASCAL Streamline 577 and the MC 500-Vixi™, and the laser method was classified into three categories: EM; single-shot using PASCAL with arbitrary energy settings (PSS-SDM; and MC500-VixiTM (VX-SDM, which were performed in eight eyes from four Dutch-Belted rabbits. In EM, 100 mW (100% was set as a landmark, and the laser energy parameters were gradually decreased to 80%, 60%, 50%, 40%, 30%, 20%, and 10%, using a 2 × 3 square pattern. In PSS-SDM and VX-SDM, as control, the laser energy was gradually decreased to 100, 80, 60, 50, 40, 30, 20, and 10 mW. The laser settings were fixed at 200 μm, 20 ms, and a wavelength of 577 μm. To identify and compare the extent of tissue damage at each spot size, optical coherence tomography (OCT and histological findings were used to construct a three-dimensional histopathology image using a confocal laser scanning fluorescence microscope.The spot size at 50% setting on EM was 7183 μm2; PSS-SDM required 50 mW (5503 μm2 to 60 mW (10279 μm2 and VX-SDM required 50 mW (7423 μm2 to create the approximate spot size. Furthermore, at 50 mW of PSS-SDM and VX-SDM, the extent of tissue damage in all three methods was generally in accord with the outer nuclear layer by OCT and inner nuclear layer by histopathological imaging.These findings suggest that it may be possible to perform subthreshold laser therapy using approximations from the EM algorithm.

  7. Laser confocal microscope for analysis of 3013 inner container closure weld region

    Energy Technology Data Exchange (ETDEWEB)

    Martinez-Rodriguez, M. J. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2017-10-26

    As part of the protocol to investigate the corrosion in the inner container closure weld region (ICCWR) a laser confocal microscope (LCM) was used to perform close visual examination of the surface and measurements of corrosion features on the surface. However, initial analysis of selected destructively evaluated (DE) containers using the LCM revealed several challenges for acquiring, processing and interpreting the data. These challenges include topography of the ICCWR sample, surface features, and the amount of surface area for collecting data at high magnification conditions. In FY17, the LCM parameters were investigated to identify the appropriate parameter values for data acquisition and identification of regions of interest. Using these parameter values, selected DE containers were analyzed to determine the extent of the ICCWR to be examined.

  8. Confocal laser endomicroscopy for diagnosis of Barrett´s esophagus

    Directory of Open Access Journals (Sweden)

    Helmut eNeumann

    2012-05-01

    Full Text Available Barrett´s esophagus (BE is established as a premalignant condition in the distal esophagus. Current surveillance guidelines recommend random biopsies every 1-2 cm at intervals of 3-5 years. Advanced endoscopic imaging of BE underwent several technical revolutions within the last decade including broad-field (red-flag techniques (e.g. chromoendoscopy and small-field techniques with confocal laser endomicroscopy (CLE at the forefront. In this review we will focus on advanced endoscopic imaging using CLE for the diagnosis and characterization of BE and associated neoplasia. In addition, we will critically discuss the technique of CLE and provide some tricks and hints for the daily routine practice of CLE for diagnosis of BE.

  9. Measurement of buried undercut structures in microfluidic devices by laser fluorescent confocal microscopy

    International Nuclear Information System (INIS)

    Li Shiguang; Liu Jing; Nguyen, Nam-Trung; Fang Zhongping; Yoon, Soon Fatt

    2009-01-01

    Measuring buried, undercut microstructures is a challenging task in metrology. These structures are usually characterized by measuring their cross sections after physically cutting the samples. This method is destructive and the obtained information is incomplete. The distortion due to cutting also affects the measurement accuracy. In this paper, we first apply the laser fluorescent confocal microscopy and intensity differentiation algorithm to obtain the complete three-dimensional profile of the buried, undercut structures in microfluidic devices, which are made by the soft lithography technique and bonded by the oxygen plasma method. The impact of material wettability and the refractive index (n) mismatch among the liquid, samples, cover layer, and objective on the measurement accuracy are experimentally investigated.

  10. Confocal laser endomicroscopy for diagnosis and histomorphologic imaging of brain tumors in vivo.

    Directory of Open Access Journals (Sweden)

    Sebastian Foersch

    Full Text Available Early detection and evaluation of brain tumors during surgery is crucial for accurate resection. Currently cryosections during surgery are regularly performed. Confocal laser endomicroscopy (CLE is a novel technique permitting in vivo histologic imaging with miniaturized endoscopic probes at excellent resolution. Aim of the current study was to evaluate CLE for in vivo diagnosis in different types and models of intracranial neoplasia. In vivo histomorphology of healthy brains and two different C6 glioma cell line allografts was evaluated in rats. One cell line expressed EYFP, the other cell line was used for staining with fluorescent dyes (fluorescein, acriflavine, FITC-dextran and Indocyanine green. To evaluate future application in patients, fresh surgical resection specimen of human intracranial tumors (n = 15 were examined (glioblastoma multiforme, meningioma, craniopharyngioma, acoustic neurinoma, brain metastasis, medulloblastoma, epidermoid tumor. Healthy brain tissue adjacent to the samples served as control. CLE yielded high-quality histomorphology of normal brain tissue and tumors. Different fluorescent agents revealed distinct aspects of tissue and cell structure (nuclear pattern, axonal pathways, hemorrhages. CLE discrimination of neoplastic from healthy brain tissue was easy to perform based on tissue and cellular architecture and resemblance with histopathology was excellent. Confocal laser endomicroscopy allows immediate in vivo imaging of normal and neoplastic brain tissue at high resolution. The technology might be transferred to scientific and clinical application in neurosurgery and neuropathology. It may become helpful to screen for tumor free margins and to improve the surgical resection of malignant brain tumors, and opens the door to in vivo molecular imaging of tumors and other neurologic disorders.

  11. Penetration pattern of rhodamine dyes into enamel and dentin: confocal laser microscopy observation.

    Science.gov (United States)

    Kwon, S R; Wertz, P W; Li, Y; Chan, D C N

    2012-02-01

    Enamel and dentin are susceptible to extrinsic and intrinsic stains. The purposes of this study were to determine the penetration pattern of Rhodamine B and dextran-conjugated Rhodamine B into the enamel and dentin as observed by confocal laser microscopy and to relate it to the penetration pattern of hydrogen peroxide commonly used as an active ingredient in tooth-whitening agents and high-molecular-weight staining molecules. Eighteen recently extracted human maxillary anterior teeth were used. Teeth were cleaned and painted with nail varnish except for the crown area above the cemento-enamel junction (CEJ). The painted teeth were then immersed in Rhodamine B and dextran-conjugated Rhodamine B (70 000 MW) for 4, 7, 10 and 15 days. Teeth were sliced to 3 mm thickness in transverse plane and mounted on a glass slide just prior to observation with confocal laser microscopy. Rhodamine B and dextran-conjugated Rhodamine B readily penetrated into the enamel and dentin when exposed for 4 and 7 days, respectively. Rhodamine B penetrated along the interprismatic spaces of the enamel into the dentin. The penetration was accentuated in sections with existing crack lines in the enamel. Rhodamine B was readily absorbed into the dentinal tubules at the dentino-enamel junction and continued to penetrate through the dentin via the dentinal tubules into the pre-dentin. Within the limitations of this study, it is concluded that Rhodamine B and dextran-conjugated Rhodamine B when applied to the external surface of the tooth readily penetrate into the enamel and dentin via the interprismatic spaces in the enamel and dentinal tubules in the dentin, suggesting that stain molecules and bleaching agents possibly exhibit similar penetration pathways. © 2011 The Authors. ICS © 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  12. High-resolution adaptive optics scanning laser ophthalmoscope with multiple deformable mirrors

    Science.gov (United States)

    Chen, Diana C.; Olivier, Scot S.; Jones; Steven M.

    2010-02-23

    An adaptive optics scanning laser ophthalmoscopes is introduced to produce non-invasive views of the human retina. The use of dual deformable mirrors improved the dynamic range for correction of the wavefront aberrations compared with the use of the MEMS mirror alone, and improved the quality of the wavefront correction compared with the use of the bimorph mirror alone. The large-stroke bimorph deformable mirror improved the capability for axial sectioning with the confocal imaging system by providing an easier way to move the focus axially through different layers of the retina.

  13. Confocal laser microscopic imaging of conspicuous facial pores in vivo: relation between the appearance and the internal structure of skin.

    Science.gov (United States)

    Sugata, Keiichi; Nishijima, Takafumi; Kitahara, Takashi; Takema, Yoshinori

    2008-05-01

    Conspicuous facial pores are one of the more serious esthetic defects of most concern to women. Previous microscopic observations of the skin surface around conspicuous pores have discovered large hollows and uneven skin tone. In this study, the observation area was extended from the skin surface to deeper skin to find the characteristic features of conspicuous pores in a wider spectrum. First, a magnified surface image of the cheek skin was obtained using a video microscope. Second, replicas were collected from the same area. Third, the horizontal cross-sectioned images of the epidermis and papillary dermis in different depths were non-invasively obtained using in vivo confocal laser scanning microscopy. These images were compared with each other to find a correlation between features of the skin surface and those of deeper layers. In cross-sectioned images of conspicuous pores, a strongly undulated epidermal-dermal junction was commonly observed around a pore's opening. Areas with this feature correlated well to the areas with larger hollows and an uneven skin tone. Our results indicate that there is a positive correlation between the incidence of the characteristic feature at the epidermal-dermal junction and the visual appearance of a pore.

  14. Theoretical investigation of confocal microscopy using an elliptically polarized cylindrical vector laser beam: Visualization of quantum emitters near interfaces

    Science.gov (United States)

    Boichenko, Stepan

    2018-04-01

    We theoretically study laser-scanning confocal fluorescence microscopy using elliptically polarized cylindrical vector excitation light as a tool for visualization of arbitrarily oriented single quantum dipole emitters located (1) near planar surfaces enhancing fluorescence, (2) in a thin supported polymer film, (3) in a freestanding polymer film, and (4) in a dielectric planar microcavity. It is shown analytically that by using a tightly focused azimuthally polarized beam, it is possible to exclude completely the orientational dependence of the image intensity maximum of a quantum emitter that absorbs light as a pair of incoherent independent linear dipoles. For linear dipole quantum emitters, the orientational independence degree higher than 0.9 can normally be achieved (this quantity equal to 1 corresponds to completely excluded orientational dependence) if the collection efficiency of the microscope objective and the emitter's total quantum yield are not strongly orientationally dependent. Thus, the visualization of arbitrarily oriented single quantum emitters by means of the studied technique can be performed quite efficiently.

  15. Achievement report for fiscal 1999 on research and development of technologies for medical welfare equipment. Chromosome image analysis and diagnosis device with confocal scanning laser microscope; 1999 nendo iryo fukushi kiki gijutsu kenkyu kaihatsu seika hokokusho. Kyoshoten laser kenbikyo ni yoru zensenshokutai gazo kaiseki shindan sochi

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-05-01

    The device is now satisfactorily capable of all image processing functions (bi-levelling, color deviation, brilliance adjustment, etc.) with the exception the karyotyping function. A substance that emits fluorescence by laser excitation is developed (for labelling the probe). A direct labelling method is studied, in which nucleic acid probes (nucleic acid labelled for chromosomal identification) will prove to be remarkably high in sensitivity. A new fluorescent reagent is synthesized, which is a BHHCT-4-dUTP to act as a nucleic acid probe. It is found that the new substance is superior to the conventional fluorescent substances in terms of stability and that all chromosomes may be forced into hybridization (selective combination of a probe with a peculiar chromosome) when various probes are labelled by this substance. A programing unit is constructed for a chromosomal aberration database. Items needed relative to chromosomes and parameters relative to chromosomal aberration data are appropriately arranged, and conditions to satisfy for their development into a database are studied. A rough design of an interface is complete. (NEDO)

  16. Evaluation of 3-D Laser Scanning Equipment : 2018 Final Report

    Science.gov (United States)

    2018-05-01

    As a follow-up to ICT Project R27-030, Evaluation of 3-D Laser Scanning, this report provides findings of an evaluation of 3-D laser scanning equipment to determine the tangible costs versus benefits and the manpower savings realized by using the equ...

  17. Structural monitoring of tunnels using terrestrial laser scanning

    NARCIS (Netherlands)

    Lindenbergh, R.C.; Uchanski, L.; Bucksch, A.; Van Gosliga, R.

    2009-01-01

    In recent years terrestrial laser scanning is rapidly evolving as a surveying technique for the monitoring of engineering objects like roof constructions, mines, dams, viaducts and tunnels. The advantage of laser scanning above traditional surveying methods is that it allows for the rapid

  18. Characterization of hydrogel microstructure using laser tweezers particle tracking and confocal reflection imaging

    International Nuclear Information System (INIS)

    Kotlarchyk, M A; Botvinick, E L; Putnam, A J

    2010-01-01

    Hydrogels are commonly used as extracellular matrix mimetics for applications in tissue engineering and increasingly as cell culture platforms with which to study the influence of biophysical and biochemical cues on cell function in 3D. In recent years, a significant number of studies have focused on linking substrate mechanical properties to cell function using standard methodologies to characterize the bulk mechanical properties of the hydrogel substrates. However, current understanding of the correlations between the microstructural mechanical properties of hydrogels and cell function in 3D is poor, in part because of a lack of appropriate techniques. Here we have utilized a laser tracking system, based on passive optical microrheology instrumentation, to characterize the microstructure of viscoelastic fibrin clots. Trajectories and mean square displacements were observed as bioinert PEGylated (PEG: polyethylene glycol) microspheres (1, 2 or 4.7 μm in diameter) diffused within confined pores created by the protein phase of fibrin hydrogels. Complementary confocal reflection imaging revealed microstructures comprised of a highly heterogeneous fibrin network with a wide range of pore sizes. As the protein concentration of fibrin gels was increased, our quantitative laser tracking measurements showed a corresponding decrease in particle mean square displacements with greater resolution and sensitivity than conventional imaging techniques. This platform-independent method will enable a more complete understanding of how changes in substrate mechanical properties simultaneously influence other microenvironmental parameters in 3D cultures.

  19. Fluorescence (Multiwave) Confocal Microscopy.

    Science.gov (United States)

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

    2016-10-01

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

  20. Cell differentiation in cardiac myxomas: confocal microscopy and gene expression analysis after laser capture microdissection.

    Science.gov (United States)

    Pucci, Angela; Mattioli, Claudia; Matteucci, Marco; Lorenzini, Daniele; Panvini, Francesca; Pacini, Simone; Ippolito, Chiara; Celiento, Michele; De Martino, Andrea; Dolfi, Amelio; Belgio, Beatrice; Bortolotti, Uberto; Basolo, Fulvio; Bartoloni, Giovanni

    2018-05-22

    Cardiac myxomas are rare tumors with a heterogeneous cell population including properly neoplastic (lepidic), endothelial and smooth muscle cells. The assessment of neoplastic (lepidic) cell differentiation pattern is rather difficult using conventional light microscopy immunohistochemistry and/or whole tissue extracts for mRNA analyses. In a preliminary study, we investigated 20 formalin-fixed and paraffin-embedded cardiac myxomas by means of conventional immunohistochemistry; in 10/20 cases, cell differentiation was also analyzed by real-time RT-PCR after laser capture microdissection of the neoplastic cells, whereas calretinin and endothelial antigen CD31 immunoreactivity was localized in 4/10 cases by double immunofluorescence confocal microscopy. Gene expression analyses of α-smooth muscle actin, endothelial CD31 antigen, alpha-cardiac actin, matrix metalloprotease-2 (MMP2) and tissue inhibitor of matrix metalloprotease-1 (TIMP1) was performed on cDNA obtained from either microdissected neoplastic cells or whole tumor sections. We found very little or absent CD31 and α-Smooth Muscle Actin expression in the microdissected cells as compared to the whole tumors, whereas TIMP1 and MMP2 genes were highly expressed in both ones, greater levels being found in patients with embolic phenomena. α-Cardiac Actin was not detected. Confocal microscopy disclosed two different signals corresponding to calretinin-positive myxoma cells and to endothelial CD31-positive cells, respectively. In conclusion, the neoplastic (lepidic) cells showed a distinct gene expression pattern and no consistent overlapping with endothelial and smooth muscle cells or cardiac myocytes; the expression of TIMP1 and MMP2 might be related to clinical presentation; larger series studies using also systematic transcriptome analysis might be useful to confirm the present results.

  1. Real time diagnosis of bladder cancer with probe-based confocal laser endomicroscopy

    Science.gov (United States)

    Liu, Jen-Jane; Wu, Katherine; Adams, Winifred; Hsiao, Shelly T.; Mach, Kathleen E.; Beck, Andrew H.; Jensen, Kristin C.; Liao, Joseph C.

    2011-02-01

    Probe-based confocal laser endomicroscopy (pCLE) is an emerging technology for in vivo optical imaging of the urinary tract. Particularly for bladder cancer, real time optical biopsy of suspected lesions will likely lead to improved management of bladder cancer. With pCLE, micron scale resolution is achieved with sterilizable imaging probes (1.4 or 2.6 mm diameter), which are compatible with standard cystoscopes and resectoscopes. Based on our initial experience to date (n = 66 patients), we have demonstrated the safety profile of intravesical fluorescein administration and established objective diagnostic criteria to differentiate between normal, benign, and neoplastic urothelium. Confocal images of normal bladder showed organized layers of umbrella cells, intermediate cells, and lamina propria. Low grade bladder cancer is characterized by densely packed monomorphic cells with central fibrovascular cores, whereas high grade cancer consists of highly disorganized microarchitecture and pleomorphic cells with indistinct cell borders. Currently, we are conducting a diagnostic accuracy study of pCLE for bladder cancer diagnosis. Patients scheduled to undergo transurethral resection of bladder tumor are recruited. Patients undergo first white light cystocopy (WLC), followed by pCLE, and finally histologic confirmation of the resected tissues. The diagnostic accuracy is determined both in real time by the operative surgeon and offline after additional image processing. Using histology as the standard, the sensitivity, specificity, positive and negative predictive value of WLC and WLC + pCLE are calculated. With additional validation, pCLE may prove to be a valuable adjunct to WLC for real time diagnosis of bladder cancer.

  2. Laser safety in design of near-infrared scanning LIDARs

    Science.gov (United States)

    Zhu, X.; Elgin, D.

    2015-05-01

    3D LIDARs (Light Detection and Ranging) with 1.5μm nanosecond pulse lasers have been increasingly used in different applications. The main reason for their popularity is that these LIDARs have high performance while at the same time can be made eye-safe. Because the laser hazard effect on eyes or skin at this wavelength region (industrial mining applications. We have incorporated the laser safety requirements in the LIDAR design and conducted laser safety analysis for different operational scenarios. While 1.5μm is normally said to be the eye-safe wavelength, in reality a high performance 3D LIDAR needs high pulse energy, small beam size and high pulse repetition frequency (PRF) to achieve long range, high resolution and high density images. The resulting radiant exposure of its stationary beam could be many times higher than the limit for a Class 1 laser device. Without carefully choosing laser and scanning parameters, including field-of-view, scan speed and pattern, a scanning LIDAR can't be eye- or skin-safe based only on its wavelength. This paper discusses the laser safety considerations in the design of eye-safe scanning LIDARs, including laser pulse energy, PRF, beam size and scanning parameters in two basic designs of scanning mechanisms, i.e. galvanometer based scanner and Risley prism based scanner. The laser safety is discussed in terms of device classification, nominal ocular hazard distance (NOHD) and safety glasses optical density (OD).

  3. Confocal microscopy imaging of the biofilm matrix

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Meyer, Rikke L

    2017-01-01

    The extracellular matrix is an integral part of microbial biofilms and an important field of research. Confocal laser scanning microscopy is a valuable tool for the study of biofilms, and in particular of the biofilm matrix, as it allows real-time visualization of fully hydrated, living specimens...... the concentration of solutes and the diffusive properties of the biofilm matrix....

  4. Clinical impact of confocal laser endomicroscopy in the management of gastrointestinal lesions with an uncertain diagnosis.

    Science.gov (United States)

    Robles-Medranda, Carlos; Vargas, Maria; Ospina, Jesenia; Puga-Tejada, Miguel; Valero, Manuel; Soria, Miguel; Bravo, Gladys; Robles-Jara, Carlos; Lukashok, Hannah Pitanga

    2017-08-16

    To evaluate the clinical impact of confocal laser endomicroscopy (CLE) in the diagnosis and management of patients with an uncertain diagnosis. A retrospective chart review was performed. Patients who underwent CLE between November 2013 and October 2015 and exhibited a poor correlation between endoscopic and histological findings were included. Baseline characteristics, indications, previous diagnostic studies, findings at the time of CLE, clinical management and histological results were analyzed. Interventions based on CLE findings were also analyzed. We compared the diagnostic accuracy of CLE and target biopsies of surgical specimens. A total of 144 patients were included. Of these, 51% (74/144) were female. The mean age was 51 years old. In all, 41/144 (28.4%) lesions were neoplastic (13 bile duct, 10 gastric, 8 esophageal, 6 colonic, 1 duodenal, 1 rectal, 1 ampulloma and 1 pancreatic). The sensitivity, specificity, positive predictive value, negative predictive value, and observed agreement when CLE was used to detect N-lesions were 85.37%, 87.38%, 72.92%, 93.75% and 86.81%, respectively. Cohen's Kappa was 69.20%, thus indicating good agreement. Changes in management were observed in 54% of the cases. CLE is a new diagnostic tool that has a significant clinical impact on the diagnosis and treatment of patients with uncertain diagnosis.

  5. Confocal laser-induced fluorescence detector for narrow capillary system with yoctomole limit of detection.

    Science.gov (United States)

    Weaver, Mitchell T; Lynch, Kyle B; Zhu, Zaifang; Chen, Huang; Lu, Joann J; Pu, Qiaosheng; Liu, Shaorong

    2017-04-01

    Laser-induced fluorescence (LIF) detectors for low-micrometer and sub-micrometer capillary on-column detection are not commercially available. In this paper, we describe in details how to construct a confocal LIF detector to address this issue. We characterize the detector by determining its limit of detection (LOD), linear dynamic range (LDR) and background signal drift; a very low LOD (~70 fluorescein molecules or 12 yoctomole fluorescein), a wide LDR (greater than 3 orders of magnitude) and a small background signal drift (~1.2-fold of the root mean square noise) are obtained. For detecting analytes inside a low-micrometer and sub-micrometer capillary, proper alignment is essential. We present a simple protocol to align the capillary with the optical system and use the position-lock capability of a translation stage to fix the capillary in position during the experiment. To demonstrate the feasibility of using this detector for narrow capillary systems, we build a 2-μm-i.d. capillary flow injection analysis (FIA) system using the newly developed LIF prototype as a detector and obtain an FIA LOD of 14 zeptomole fluorescein. We also separate a DNA ladder sample by bare narrow capillary - hydrodynamic chromatography and use the LIF prototype to monitor the resolved DNA fragments. We obtain not only well-resolved peaks but also the quantitative information of all DNA fragments. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. A laser sheet self-calibration method for scanning PIV

    Science.gov (United States)

    Knutsen, Anna N.; Lawson, John M.; Dawson, James R.; Worth, Nicholas A.

    2017-10-01

    Knowledge of laser sheet position, orientation, and thickness is a fundamental requirement of scanning PIV and other laser-scanning methods. This paper describes the development and evaluation of a new laser sheet self-calibration method for stereoscopic scanning PIV, which allows the measurement of these properties from particle images themselves. The approach is to fit a laser sheet model by treating particles as randomly distributed probes of the laser sheet profile, whose position is obtained via a triangulation procedure enhanced by matching particle images according to their variation in brightness over a scan. Numerical simulations and tests with experimental data were used to quantify the sensitivity of the method to typical experimental error sources and validate its performance in practice. The numerical simulations demonstrate the accurate recovery of the laser sheet parameters over range of different seeding densities and sheet thicknesses. Furthermore, they show that the method is robust to significant image noise and camera misalignment. Tests with experimental data confirm that the laser sheet model can be accurately reconstructed with no impairment to PIV measurement accuracy. The new method is more efficient and robust in comparison with the standard (self-) calibration approach, which requires an involved, separate calibration step that is sensitive to experimental misalignments. The method significantly improves the practicality of making accurate scanning PIV measurements and broadens its potential applicability to scanning systems with significant vibrations.

  7. Maritime Laser Scanning as the Source for Spatial Data

    Directory of Open Access Journals (Sweden)

    Szulwic Jakub

    2015-12-01

    Full Text Available The rapid development of scanning technology, especially mobile scanning, gives the possibility to collect spatial data coming from maritime measurement platforms and autonomous manned or unmanned vehicles. Presented solution is derived from the mobile scanning. However we should keep in mind that the specificity of laser scanning at sea and processing collected data should be in the form acceptable in Geographical Information Systems, especially typical for the maritime needs. At the same time we should be aware that data coming from maritime mobile scanning constitutes a new approach to the describing of maritime environment and brings a new perspective that is completely different than air and terrestrial scanning.

  8. Power Measurements for Microvision, Inc., Aircrew Integrated Helmet System Scanning Laser Helmet-Mounted Display

    National Research Council Canada - National Science Library

    Rash, Clarence

    2002-01-01

    ...) technology based on scanning lasers. Under this program, Microvision, Inc., Bothell, Washington, has developed a scanning laser HMD prototype for use with the Aircrew Integrated Helmet System (AIHS...

  9. Identification of nodal tissue in the living heart using rapid scanning fiber-optics confocal microscopy and extracellular fluorophores.

    Science.gov (United States)

    Huang, Chao; Kaza, Aditya K; Hitchcock, Robert W; Sachse, Frank B

    2013-09-01

    Risks associated with pediatric reconstructive heart surgery include injury of the sinoatrial node (SAN) and atrioventricular node (AVN), requiring cardiac rhythm management using implantable pacemakers. These injuries are the result of difficulties in identifying nodal tissues intraoperatively. Here we describe an approach based on confocal microscopy and extracellular fluorophores to quantify tissue microstructure and identify nodal tissue. Using conventional 3-dimensional confocal microscopy we investigated the microstructural arrangement of SAN, AVN, and atrial working myocardium (AWM) in fixed rat heart. AWM exhibited a regular striated arrangement of the extracellular space. In contrast, SAN and AVN had an irregular, reticulated arrangement. AWM, SAN, and AVN tissues were beneath a thin surface layer of tissue that did not obstruct confocal microscopic imaging. Subsequently, we imaged tissues in living rat hearts with real-time fiber-optics confocal microscopy. Fiber-optics confocal microscopy images resembled images acquired with conventional confocal microscopy. We investigated spatial regularity of tissue microstructure from Fourier analysis and second-order image moments. Fourier analysis of fiber-optics confocal microscopy images showed that the spatial regularity of AWM was greater than that of nodal tissues (37.5 ± 5.0% versus 24.3 ± 3.9% for SAN and 23.8 ± 3.7% for AVN; Pfiber-optics confocal microscopy. Application of the approach in pediatric reconstructive heart surgery may reduce risks of injuring nodal tissues.

  10. scanning speed influence on the physical properties of laser metal

    African Journals Online (AJOL)

    user

    2017-01-01

    Jan 1, 2017 ... result of the preliminary study that produces full dense and pore free deposits. ... Keywords: Additive manufacturing, Laser metal deposition (LMD), Material efficiency, Titanium alloy. 1. ... parts. Ti6Al4V is the most commonly produced titanium alloy ... In this study, effect of laser transverse speed or scanning.

  11. Multicolor pattern scan laser for diabetic retinopathy with cataract

    Institute of Scientific and Technical Information of China (English)

    Takao; Hirano; Yasuhiro; Iesato; Toshinori; Murata

    2014-01-01

    · AIM: To evaluate the ability of various laser wavelengths in delivering sufficient burns to the retina in eyes with cataract using a new multicolor pattern scan laser with green(532 nm), yellow(577 nm), and red(647 nm)lasers.·METHODS: The relationship between the Emery-Little(EL) degree of cataract severity and the laser wavelength required to deliver adequate burns was investigated in102 diabetic eyes. Treatment time, total number of laser shots, and intra-operative pain were assessed as well.·RESULTS: All EL-1 grade eyes and 50% of EL-2 eyes were successfully treated with the green laser, while 50%of EL-2 eyes, 96% of EL-3 eyes, and 50% of EL-4 eyes required the yellow laser. The red laser was effective in the remaining 4% of EL-3 and 50% of EL-4 eyes.·CONCLUSION: Longer wavelength lasers are more effective in delivering laser burns through cataract when we use a multicolor pattern scan laser system.

  12. Monitoring pancreatic carcinogenesis by the molecular imaging of cathepsin E in vivo using confocal laser endomicroscopy.

    Directory of Open Access Journals (Sweden)

    Hui Li

    Full Text Available The monitoring of pancreatic ductal adenocarcinoma (PDAC in high-risk populations is essential. Cathepsin E (CTSE is specifically and highly expressed in PDAC and pancreatic intraepithelial neoplasias (PanINs, and its expression gradually increases along with disease progression. In this study, we first established an in situ 7,12-dimethyl-1,2-benzanthracene (DMBA-induced rat model for PanINs and PDAC and then confirmed that tumorigenesis properties in this model were consistent with those of human PDAC in that CTSE expression gradually increased with tumor development using histology and immunohistochemistry. Then, using in vivo imaging of heterotopically implanted tumors generated from CTSE- overexpressing cells (PANC-1-CTSE in nude mice and in vitro imaging of PanINs and PDAC in DMBA-induced rats, the specificity of the synthesized CTSE-activatable probe was verified. Quantitative determination identified that the fluorescence signal ratio of pancreatic tumor to normal pancreas gradually increased in association with progressive pathological grades, with the exception of no significant difference between PanIN-II and PanIN-III grades. Finally, we monitored pancreatic carcinogenesis in vivo using confocal laser endomicroscopy (CLE in combination with the CTSE-activatable probe. A prospective double-blind control study was performed to evaluate the accuracy of this method in diagnosing PDAC and PanINs of all grades (>82.7%. This allowed us to establish effective diagnostic criteria for CLE in PDAC and PanINs to facilitate the monitoring of PDAC in high-risk populations.

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

    Science.gov (United States)

    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...

  14. Facial recognition and laser surface scan: a pilot study

    DEFF Research Database (Denmark)

    Lynnerup, Niels; Clausen, Maja-Lisa; Kristoffersen, Agnethe May

    2009-01-01

    Surface scanning of the face of a suspect is presented as a way to better match the facial features with those of a perpetrator from CCTV footage. We performed a simple pilot study where we obtained facial surface scans of volunteers and then in blind trials tried to match these scans with 2D...... photographs of the faces of the volunteers. Fifteen male volunteers were surface scanned using a Polhemus FastSCAN Cobra Handheld Laser Scanner. Three photographs were taken of each volunteer's face in full frontal, profile and from above at an angle of 45 degrees and also 45 degrees laterally. Via special...

  15. Near-Infrared Confocal Laser Reflectance Cytoarchitectural Imaging of the Substantia Nigra and Cerebellum in the Fresh Human Cadaver.

    Science.gov (United States)

    Cheyuo, Cletus; Grand, Walter; Balos, Lucia L

    2017-01-01

    Cytoarchitectural neuroimaging remains critical for diagnosis of many brain diseases. Fluorescent dye-enhanced, near-infrared confocal in situ cellular imaging of the brain has been reported. However, impermeability of the blood-brain barrier to most fluorescent dyes limits clinical utility of this modality. The differential degree of reflectance from brain tissue with unenhanced near-infrared imaging may represent an alternative technique for in situ cytoarchitectural neuroimaging. We assessed the utility of unenhanced near-infrared confocal laser reflectance imaging of the cytoarchitecture of the cerebellum and substantia nigra in 2 fresh human cadaver brains using a confocal near-infrared laser probe. Cellular images based on near-infrared differential reflectance were captured at depths of 20-180 μm from the brain surface. Parts of the cerebellum and substantia nigra imaged using the probe were subsequently excised and stained with hematoxylin and eosin for histologic correlation. Near-infrared reflectance imaging revealed the 3-layered cytoarchitecture of the cerebellum, with Purkinje cells appearing hyperreflectant. In the substantia nigra, neurons appeared hyporeflectant with hyperreflectant neuromelanin cytoplasmic inclusions. Cytoarchitecture of the cerebellum and substantia nigra revealed on near-infrared imaging closely correlated with the histology on hematoxylin-eosin staining. We showed that unenhanced near-infrared reflectance imaging of fresh human cadaver brain can reliably identify and distinguish neurons and detailed cytoarchitecture of the cerebellum and substantia nigra. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Concurrent Reflectance Confocal Microscopy and Laser Doppler Flowmetry to Improve Skin Cancer Imaging: A Monte Carlo Model and Experimental Validation

    Directory of Open Access Journals (Sweden)

    Alireza Mowla

    2016-09-01

    Full Text Available Optical interrogation of suspicious skin lesions is standard care in the management of skin cancer worldwide. Morphological and functional markers of malignancy are often combined to improve expert human diagnostic power. We propose the evaluation of the combination of two independent optical biomarkers of skin tumours concurrently. The morphological modality of reflectance confocal microscopy (RCM is combined with the functional modality of laser Doppler flowmetry, which is capable of quantifying tissue perfusion. To realize the idea, we propose laser feedback interferometry as an implementation of RCM, which is able to detect the Doppler signal in addition to the confocal reflectance signal. Based on the proposed technique, we study numerical models of skin tissue incorporating two optical biomarkers of malignancy: (i abnormal red blood cell velocities and concentrations and (ii anomalous optical properties manifested through tissue confocal reflectance, using Monte Carlo simulation. We also conduct a laboratory experiment on a microfluidic channel containing a dynamic turbid medium, to validate the efficacy of the technique. We quantify the performance of the technique by examining a signal to background ratio (SBR in both the numerical and experimental models, and it is shown that both simulated and experimental SBRs improve consistently using this technique. This work indicates the feasibility of an optical instrument, which may have a role in enhanced imaging of skin malignancies.

  17. Evolution of laser skin resurfacing: from scanning to fractional technology.

    Science.gov (United States)

    Aslam, Arif; Alster, Tina S

    2014-11-01

    Laser skin resurfacing was popularized for photoaged and scarred skin 2 decades ago. Since then, several technologic advancements have led to a new generation of delivery systems that produce excellent clinical outcomes with reduced treatment risks and faster recovery times. To review the evolution of laser skin resurfacing from pulsed and scanned infrared laser technology to the latest techniques of nonablative and ablative fractional photothermolysis. All published literature regarding laser skin resurfacing was analyzed and collated. A comprehensive review of laser skin resurfacing was outlined and future developments in the field of fractionated laser skin treatment were introduced. Laser skin resurfacing has evolved such that excellent clinical outcomes in photodamaged and scarred skin are achieved with rapid wound healing. As newer devices are developed, the applications of this technology will have a dramatic effect on the delivery of medical and aesthetic dermatology.

  18. Influence of laser frequency noise on scanning Fabry-Perot interferometer based laser Doppler velocimetry

    DEFF Research Database (Denmark)

    Rodrigo, Peter John; Pedersen, Christian

    2014-01-01

    n this work, we study the performance of a scanning Fabry-Perot interferometer based laser Doppler velocimeter (sFPILDV) and compare two candidate 1.5 um single-frequency laser sources for the system – a fiber laser (FL) and a semiconductor laser (SL). We describe a straightforward calibration...... procedure for the sFPI-LDV and investigate the effect of different degrees of laser frequency noise between the FL and the SL on the velocimeter’s performance...

  19. Modifying a Rodenstock scanning laser ophthalmoscope for imaging densitometry.

    Science.gov (United States)

    Tornow, R P; Beuel, S; Zrenner, E

    1997-08-01

    The necessary modifications and technical requirements are described for using a commercially available scanning laser ophthalmoscope (Rodenstock Model 101 SLO) as an imaging densitometer to assess human photopigment distribution. The main requirements are a linear detector amplifier, fast shutters for the laser beams, and a trigger unit. Images must be compensated for varying laser intensity. Both rod and cone photopigments are measured with the 514-nm argon laser of the SLO. Discrimination is possible owing to the different spatial distribution. The cone pigment density peaks in the foveal center (D = 0.40) with a steep decrease with increasing eccentricity E (full width at half-maximum, 2.5 degrees ). Rod photopigment increases with increasing eccentricity (D = 0.23 for E = 11 degrees ). These values are in agreement with previous reported results obtained with scanning laser ophthalmoscopes specially designed for retinal densitometry and high stability.

  20. Accuracy of a digital impression system based on parallel confocal laser technology for implants with consideration of operator experience and implant angulation and depth.

    Science.gov (United States)

    Giménez, Beatriz; Özcan, Mutlu; Martínez-Rus, Francisco; Pradíes, Guillermo

    2014-01-01

    To evaluate the accuracy of a digital impression system based on parallel confocal red laser technology, taking into consideration clinical parameters such as operator experience and angulation and depth of implants. A maxillary master model with six implants (located bilaterally in the second molar, second premolar, and lateral incisor positions) was fitted with six polyether ether ketone scan bodies. One second premolar implant was placed with 30 degrees of mesial angulation; the opposite implant was positioned with 30 degrees of distal angulation. The lateral incisor implants were placed 2 or 4 mm subgingivally. Two experienced and two inexperienced operators performed intraoral scanning. Five different interimplant distances were then measured. The files obtained from the scans were imported with reverse-engineering software. Measurements were then made with a coordinate measurement machine, with values from the master model used as reference values. The deviations from the actual values were then calculated. The differences between experienced and inexperienced operators and the effects of different implant angulations and depths were compared statistically. Overall, operator 3 obtained significantly less accurate results. The angulated implants did not significantly influence accuracy compared to the parallel implants. Differences were found in the amount of error in the different quadrants. The second scanned quadrant had significantly worse results than the first scanned quadrant. Impressions of the implants placed at the tissue level were less accurate than implants placed 2 and 4 mm subgingivally. The operator affected the accuracy of measurements, but the performance of the operator was not necessarily dependent on experience. Angulated implants did not decrease the accuracy of the digital impression system tested. The scanned distance affected the predictability of the accuracy of the scanner, and the error increased with the increased length of the

  1. Diffractive elements performance in chromatic confocal microscopy

    International Nuclear Information System (INIS)

    Garzon, J; Duque, D; Alean, A; Toledo, M; Meneses, J; Gharbi, T

    2011-01-01

    The Confocal Laser Scanning Microscopy (CLSM) has been widely used in the semiconductor industry and biomedicine because of its depth discrimination capability. Subsequent to this technique has been developed in recent years Chromatic Confocal Microscopy. This method retains the same principle of confocal and offers the added advantage of removing the axial movement of the moving system. This advantage is usually accomplished with an optical element that generates a longitudinal chromatic aberration and a coding system that relates the axial position of each point of the sample with the wavelength that is focused on each. The present paper shows the performance of compact chromatic confocal microscope when some different diffractive elements are used for generation of longitudinal chromatic aberration. Diffractive elements, according to the process and manufacturing parameters, may have different diffraction efficiency and focus a specific wavelength in a specific focal position. The performance assessment is carried out with various light sources which exhibit an incoherent behaviour and a broad spectral width.

  2. 3D Laser Scanning in Technology Education.

    Science.gov (United States)

    Flowers, Jim

    2000-01-01

    A three-dimensional laser scanner can be used as a tool for design and problem solving in technology education. A hands-on experience can enhance learning by captivating students' interest and empowering them with creative tools. (Author/JOW)

  3. BENCHMARKING MOBILE LASER SCANNING SYSTEMS USING A PERMANENT TEST FIELD

    Directory of Open Access Journals (Sweden)

    H. Kaartinen

    2012-07-01

    Full Text Available The objective of the study was to benchmark the geometric accuracy of mobile laser scanning (MLS systems using a permanent test field under good coverage of GNSS. Mobile laser scanning, also called mobile terrestrial laser scanning, is currently a rapidly developing area in laser scanning where laser scanners, GNSS and IMU are mounted onboard a moving vehicle. MLS can be considered to fill the gap between airborne and terrestrial laser scanning. Data provided by MLS systems can be characterized with the following technical parameters: a point density in the range of 100-1000 points per m2 at 10 m distance, b distance measurement accuracy of 2-5 cm, and c operational scanning range from 1 to 100 m. Several commercial, including e.g. Riegl, Optech and others, and some research mobile laser scanning systems surveyed the test field using predefined driving speed and directions. The acquired georeferenced point clouds were delivered for analyzing. The geometric accuracy of the point clouds was determined using the reference targets that could be identified and measured from the point cloud. Results show that in good GNSS conditions most systems can reach an accuracy of 2 cm both in plane and elevation. The accuracy of a low cost system, the price of which is less than tenth of the other systems, seems to be within a few centimetres at least in ground elevation determination. Inaccuracies in the relative orientation of the instruments lead to systematic errors and when several scanners are used, in multiple reproductions of the objects. Mobile laser scanning systems can collect high density point cloud data with high accuracy. A permanent test field suits well for verifying and comparing the performance of different mobile laser scanning systems. The accuracy of the relative orientation between the mapping instruments needs more attention. For example, if the object is seen double in the point cloud due to imperfect boresight calibration between two

  4. HOVE-Wedge-Filtering of Geomorphologic Terrestrial Laser Scan Data

    Directory of Open Access Journals (Sweden)

    Helmut Panholzer

    2018-02-01

    Full Text Available Terrestrial laser scanning has become an important surveying technique in many fields such as natural hazard assessment. To analyse earth surface processes, it is useful to generate a digital terrain model originated from laser scan point cloud data. To determine the terrain surface as precisely as possible, it is often necessary to filter out points that do not represent the terrain surface. Examples are vegetation, vehicles, and animals. In mountainous terrain with a small-structured topography, filtering is very difficult. Here, automatic filtering solutions usually designed for airborne laser scan data often lead to unsatisfactory results. In this work, we further develop an existing approach for automated filtering of terrestrial laser scan data, which is based on the assumption that no other surface point can be located in the area above a direct line of sight between scanner and another measured point. By taking into account several environmental variables and a repetitive calculation method, the modified method leads to significantly better results. The root-mean-square-error (RSME for the same test measurement area could be reduced from 5.284 to 1.610. In addition, a new approach for filtering and interpolation of terrestrial laser scanning data is presented using a grid with horizontal and vertical angular data and the measurement length.

  5. In situ laser processing in a scanning electron microscope

    Energy Technology Data Exchange (ETDEWEB)

    Roberts, Nicholas A.; Magel, Gregory A.; Hartfield, Cheryl D.; Moore, Thomas M.; Fowlkes, Jason D.; Rack, Philip D. [Department of Materials Science and Engineering, University of Tennessee, Knoxville, Tennessee 37996 (United States) and Omniprobe, Inc., an Oxford Instruments Company, 10410 Miller Rd., Dallas, Texas 75238 (United States); Omniprobe, Inc., an Oxford Instruments Company, 10410 Miller Rd., Dallas, Texas 75238 (United States); Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831 (United States); Department of Materials Science and Engineering, University of Tennessee, Knoxville, Tennessee 37996 (United States) and Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831 (United States)

    2012-07-15

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

  6. Computer Aided Diagnosis for Confocal Laser Endomicroscopy in Advanced Colorectal Adenocarcinoma.

    Directory of Open Access Journals (Sweden)

    Daniela Ştefănescu

    Full Text Available Confocal laser endomicroscopy (CLE is becoming a popular method for optical biopsy of digestive mucosa for both diagnostic and therapeutic procedures. Computer aided diagnosis of CLE images, using image processing and fractal analysis can be used to quantify the histological structures in the CLE generated images. The aim of this study is to develop an automatic diagnosis algorithm of colorectal cancer (CRC, based on fractal analysis and neural network modeling of the CLE-generated colon mucosa images.We retrospectively analyzed a series of 1035 artifact-free endomicroscopy images, obtained during CLE examinations from normal mucosa (356 images and tumor regions (679 images. The images were processed using a computer aided diagnosis (CAD medical imaging system in order to obtain an automatic diagnosis. The CAD application includes image reading and processing functions, a module for fractal analysis, grey-level co-occurrence matrix (GLCM computation module, and a feature identification module based on the Marching Squares and linear interpolation methods. A two-layer neural network was trained to automatically interpret the imaging data and diagnose the pathological samples based on the fractal dimension and the characteristic features of the biological tissues.Normal colon mucosa is characterized by regular polyhedral crypt structures whereas malignant colon mucosa is characterized by irregular and interrupted crypts, which can be diagnosed by CAD. For this purpose, seven geometric parameters were defined for each image: fractal dimension, lacunarity, contrast correlation, energy, homogeneity, and feature number. Of the seven parameters only contrast, homogeneity and feature number were significantly different between normal and cancer samples. Next, a two-layer feed forward neural network was used to train and automatically diagnose the malignant samples, based on the seven parameters tested. The neural network operations were cross

  7. Scanning Laser Infrared Molecular Spectrometer (SLIMS)

    Science.gov (United States)

    Scott, David C.; Rickey, Kelly; Ksendzov, Alexander; George, Warren P.; Aljabri, Abdullah S.; Steinkraus, Joel M.

    2012-01-01

    This prototype innovation is a novel design that achieves very long, effective laser path lengths that are able to yield ppb (parts per billion) and sub-ppb measurements of trace gases. SLIMS can also accommodate multiple laser channels covering a wide range of wavelengths, resulting in detection of more chemicals of interest. The mechanical design of the mirror cell allows for the large effective path length within a small footprint. The same design provides a robust structure that lends itself to being immune to some of the alignment challenges that similar cells face. By taking a hollow cylinder and by cutting an elliptically or spherically curved surface into its inner wall, the basic geometry of a reflecting ring is created. If the curved, inner surface is diamond-turned and highly polished, a surface that is very highly reflective can be formed. The surface finish can be further improved by adding a thin chrome or gold film over the surface. This creates a high-quality, curved, mirrored surface. A laser beam, which can be injected from a small bore hole in the wall of the cylinder, will be able to make many low-loss bounces around the ring, creating a large optical path length. The reflecting ring operates on the same principle as the Herriott cell. The difference exists in the mirror that doesn't have to be optically aligned, and which has a relatively large, internal surface area that lends itself to either open air or evacuated spectroscopic measurements. This solid, spherical ring mirror removes the possibility of mirror misalignment caused by thermal expansion or vibrations, because there is only a single, solid reflecting surface. Benefits of the reflecting ring come into play when size constraints reduce the size of the system, especially for space missions in which mass is at a premium.

  8. Bus bays inventory using a terrestrial laser scanning system

    Directory of Open Access Journals (Sweden)

    Bobkowska Katarzyna

    2017-01-01

    Full Text Available This article presents the use of laser scanning technology for the assessment of bus bay geo-location. Ground laser scanning is an effective tool for collecting three-dimensional data. Moreover, the analysis of a point cloud dataset can be a source of a lot of information. The authors have outlined an innovative use of data collection and analysis using the TLS regarding information on the flatness of bus bays. The results were finalized in the form of colour three-dimensional maps of deviations and pavement type.

  9. Using a laser scanning camera for reactor inspection

    International Nuclear Information System (INIS)

    Armour, I.A.; Adrain, R.S.; Klewe, R.C.

    1984-01-01

    Inspection of nuclear reactors is normally carried out using TV or film cameras. There are, however, several areas where these cameras show considerable shortcomings. To overcome these difficulties, laser scanning cameras have been developed. This type of camera can be used for general visual inspection as well as the provision of high resolution video images with high ratio on and off-axis zoom capability. In this paper, we outline the construction and operation of a laser scanning camera and give examples of how it has been used in various power stations, and indicate future potential developments. (author)

  10. Analysis of the melanin distribution in different ethnic groups by in vivo laser scanning microscopy

    International Nuclear Information System (INIS)

    Antoniou, C; Lademann, J; Richter, H; Patzelt, A; Sterry, W; Astner, S; Zastrow, L; Koch, S

    2009-01-01

    The aim of this study was to determine whether Laser scanning confocal microscopy (LSM) is able to visualize differences in melanin content and distribution in different Skin Phototypes. The investigations were carried out on six healthy volunteers with Skin Phototypes II, IV, and VI. Representative skin samples of Skin Phototypes II, V, and VI were obtained for histological analysis from remaining tissue of skin grafts and were used for LSM-pathologic correlation. LSM evaluation showed significant differences in melanin distribution in Skin Phototypes II, IV, and VI, respectively. Based on the differences in overall reflectivity and image brightness, a visual evaluation scheme showed increasing brightness of the basal and suprabasal layers with increasing Skin Phototypes. The findings correlated well with histological analysis. The results demonstrate that LSM may serve as a promising adjunctive tool for real time assessment of melanin content and distribution in human skin, with numerous clinical applications and therapeutic and preventive implications

  11. Fluence scan: an unexplored property of a laser beam

    International Nuclear Information System (INIS)

    Chalupsky, Jaromir; Hajkova, Vera; Burian, Tomas; Juha, Libor; Polcar, Tomas; Gaudin, Jerome; Nagasono, Mitsuru; Yabashi, Makina; Sobierajski, Ryszard; Krzywinski, Jacek

    2013-01-01

    We present an extended theoretical background of so-called fluence scan (f-scan or F-scan) method, which is frequently being used for offline characterization of focused short-wavelength (EUV, soft X-ray, and hard X-ray) laser beams [J. Chalupsky et al., Opt. Express 18, 27836 (2010)]. The method exploits ablative imprints in various solids to visualize iso-fluence beam contours at different fluence and/or clip levels. An f-scan curve (clip level as a function of the corresponding iso-fluence contour area) can be generated for a general non-Gaussian beam. As shown in this paper, fluence scan encompasses important information about energy distribution within the beam profile, which may play an essential role in laser-matter interaction research employing intense non-ideal beams. Here we for the first time discuss fundamental properties of the f-scan function and its inverse counterpart (if-scan). Furthermore, we extensively elucidate how it is related to the effective beam area, energy distribution, and to the so called Liu's dependence [J.M. Liu, Opt. Lett. 7, 196 (1982)]. A new method of the effective area evaluation based on weighted inverse f-scan fit is introduced and applied to real data obtained at the SCSS (SPring-8 Compact SASE Source) facility. (authors)

  12. The learning curve, interobserver, and intraobserver agreement of endoscopic confocal laser endomicroscopy in the assessment of mucosal barrier defects.

    Science.gov (United States)

    Chang, Jeff; Ip, Matthew; Yang, Michael; Wong, Brendon; Power, Theresa; Lin, Lisa; Xuan, Wei; Phan, Tri Giang; Leong, Rupert W

    2016-04-01

    Confocal laser endomicroscopy can dynamically assess intestinal mucosal barrier defects and increased intestinal permeability (IP). These are functional features that do not have corresponding appearance on histopathology. As such, previous pathology training may not be beneficial in learning these dynamic features. This study aims to evaluate the diagnostic accuracy, learning curve, inter- and intraobserver agreement for identifying features of increased IP in experienced and inexperienced analysts and pathologists. A total of 180 endoscopic confocal laser endomicroscopy (Pentax EC-3870FK; Pentax, Tokyo, Japan) images of the terminal ileum, subdivided into 6 sets of 30 were evaluated by 6 experienced analysts, 13 inexperienced analysts, and 2 pathologists, after a 30-minute teaching session. Cell-junction enhancement, fluorescein leak, and cell dropout were used to represent increased IP and were either present or absent in each image. For each image, the diagnostic accuracy, confidence, and quality were assessed. Diagnostic accuracy was significantly higher for experienced analysts compared with inexperienced analysts from the first set (96.7% vs 83.1%, P 0.86 for experienced observers. Features representative of increased IP can be rapidly learned with high inter- and intraobserver agreement. Confidence and image quality were significant predictors of accurate interpretation. Previous pathology training did not have an effect on learning. Copyright © 2016 American Society for Gastrointestinal Endoscopy. Published by Elsevier Inc. All rights reserved.

  13. USE OF LASER SCANNING FOR CULTURAL HERITAGE DOCUMENTATION

    Directory of Open Access Journals (Sweden)

    Gulhan BENLI

    2013-01-01

    Full Text Available In terms of raising an awareness of the historical, national and cultural properties in our country and ensuring a transfer of information to posterity, it is of vital importance to take inventory of the cultural and natural real properties located in protected sites. Many fields, such as medical science, construction, ground engineering, geodetic engineering, and architecture, make use of the present-day laser scanning technology. Even if contemporary and current scientific methods are used for the inventory and documentation studies related to cultural and natural real properties in the PROTECTED SITES in the field of architecture; acquiring data of the entirety of a protected site using these methods is a time consuming process. Among the scientific methods applied, laser scanning technology has the utmost importance in the latest years. The laser scanning devices for the detection of cultural, natural and historical properties in archeological, historical, urban or mixed protected sites in Turkey, eliminate challenges such as the enormity of sites, the difficulty of working in the sites, intense work hours, and the necessity of having a thorough knowledge of the site. In the scope of this study, the usage, application, facilities, advantages and attainments of geodetic laser scanning systems in conducting surveys on facade, street or avenue silhouettes in the protected sites, where historical buildings within field of architecture are widespread, will be examined.

  14. Data acquisition considerations for Terrestrial Laser Scanning of forest plots

    NARCIS (Netherlands)

    Wilkes, Phil; Lau Sarmiento, Alvaro; Disney, Mathias; Calders, Kim; Burt, Andrew; Gonzalez De Tanago Meñaca, J.; Bartholomeus, Harm; Brede, Benjamin; Herold, Martin

    2017-01-01

    The poor constraint of forest Above Ground Biomass (AGB) is responsible, in part, for large uncertainties in modelling future climate scenarios. Terrestrial Laser Scanning (TLS) can be used to derive unbiased and non-destructive estimates of tree structure and volume and can, therefore, be used to

  15. Accuracy assessment of airborne laser scanning strips using planar features

    NARCIS (Netherlands)

    Soudarissanane, S.S.; Van der Sande, C.J.; Khoshelham, K.

    2010-01-01

    Airborne Laser Scanning (ALS) is widely used in many applications for its high measurement accuracy, fast acquisition capability, and large spatial coverage. Accuracy assessment of the ALS data usually relies on comparing corresponding tie elements, often points or lines, in the overlapping strips.

  16. Single scan vector prediction in selective laser melting

    NARCIS (Netherlands)

    Wits, Wessel Willems; Bruins, R.; Terpstra, L.; Huls, R.A.; Geijselaers, Hubertus J.M.

    2015-01-01

    In selective laser melting (SLM) products are built by melting layers of metal powder successively. Optimal process parameters are usually obtained by scanning single vectors and subsequently determining which settings lead to a good compromise between product density and build speed. This paper

  17. COMPARISON OF RETINAL PATHOLOGY VISUALIZATION IN MULTISPECTRAL SCANNING LASER IMAGING.

    Science.gov (United States)

    Meshi, Amit; Lin, Tiezhu; Dans, Kunny; Chen, Kevin C; Amador, Manuel; Hasenstab, Kyle; Muftuoglu, Ilkay Kilic; Nudleman, Eric; Chao, Daniel; Bartsch, Dirk-Uwe; Freeman, William R

    2018-03-16

    To compare retinal pathology visualization in multispectral scanning laser ophthalmoscope imaging between the Spectralis and Optos devices. This retrospective cross-sectional study included 42 eyes from 30 patients with age-related macular degeneration (19 eyes), diabetic retinopathy (10 eyes), and epiretinal membrane (13 eyes). All patients underwent retinal imaging with a color fundus camera (broad-spectrum white light), the Spectralis HRA-2 system (3-color monochromatic lasers), and the Optos P200 system (2-color monochromatic lasers). The Optos image was cropped to a similar size as the Spectralis image. Seven masked graders marked retinal pathologies in each image within a 5 × 5 grid that included the macula. The average area with detected retinal pathology in all eyes was larger in the Spectralis images compared with Optos images (32.4% larger, P < 0.0001), mainly because of better visualization of epiretinal membrane and retinal hemorrhage. The average detection rate of age-related macular degeneration and diabetic retinopathy pathologies was similar across the three modalities, whereas epiretinal membrane detection rate was significantly higher in the Spectralis images. Spectralis tricolor multispectral scanning laser ophthalmoscope imaging had higher rate of pathology detection primarily because of better epiretinal membrane and retinal hemorrhage visualization compared with Optos bicolor multispectral scanning laser ophthalmoscope imaging.

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

    Energy Technology Data Exchange (ETDEWEB)

    Darvin, M E; Richter, H; Zhu, Y J; Meinke, M C; Knorr, F; Lademann, J [Center of Experimental and Applied Cutaneous Physiology, Department of Dermatology, Venerology and Allergology, Charité - Universitätsmedizin Berlin (Germany); Gonchukov, S A [National Research Nuclear University ' ' MEPhI' ' (Russian Federation); Koenig, K [JenLab GmbH, Schillerstr. 1, 07745 Jena (Germany)

    2014-07-31

    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. (laser biophotonics)

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

    International Nuclear Information System (INIS)

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

    2014-01-01

    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. (laser biophotonics)

  20. Preliminary testing of the Scanning Laser Environmental Airborne Fluorosensor

    International Nuclear Information System (INIS)

    Brown, C.E.; Marois, R.; Fingas, M.F.; Mullin, J.V.

    2000-01-01

    The installation and testing program of the Scanning Laser Environmental Airborne Fluorosensor (SLEAF) on Environment Canada's DC-3 aircraft was described and the capabilities of the new system were presented. SLEAF is a new generation of laser fluorosensor designed to provide prompt reliable detection and mapping of oil pollution in different marine and terrestrial environments. It consists of a high-power excimer laser, high-resolution range-gated intensified diode-array spectrometer, and a pair of variable speed and angular displacement scanning mirrors. SLEAF is capable of detecting narrow bands of oil that can pile up along the high tide lines of beaches and shorelines, including those that contain ice and snow. It also has the added benefit of providing real-time detection. SLEAF will be declared operational for emergency response personnel when the initial test flight program will be completed in the near future. 9 refs., 2 figs

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

    Science.gov (United States)

    Trägårdh, J; Macrae, K; Travis, C; Amor, R; Norris, G; Wilson, S H; Oppo, G-L; McConnell, G

    2015-07-01

    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. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  2. Scanning laser optical tomography for in toto imaging of the murine cochlea.

    Directory of Open Access Journals (Sweden)

    Lena Nolte

    Full Text Available The mammalian cochlea is a complex macroscopic structure due to its helical shape and the microscopic arrangements of the individual layers of cells. To improve the outcomes of hearing restoration in deaf patients, it is important to understand the anatomic structure and composition of the cochlea ex vivo. Hitherto, only one histological technique based on confocal laser scanning microscopy and optical clearing has been developed for in toto optical imaging of the murine cochlea. However, with a growing size of the specimen, e.g., human cochlea, this technique reaches its limitations. Here, we demonstrate scanning laser optical tomography (SLOT as a valuable imaging technique to visualize the murine cochlea in toto without any physical slicing. This technique can also be applied in larger specimens up to cm3 such as the human cochlea. Furthermore, immunolabeling allows visualization of inner hair cells (otoferlin or spiral ganglion cells (neurofilament within the whole cochlea. After image reconstruction, the 3D dataset was used for digital segmentation of the labeled region. As a result, quantitative analysis of position, length and curvature of the labeled region was possible. This is of high interest in order to understand the interaction of cochlear implants (CI and cells in more detail.

  3. Fluence scan: an unexplored property of a laser beam

    Czech Academy of Sciences Publication Activity Database

    Chalupský, Jaromír; Burian, Tomáš; Hájková, Věra; Juha, Libor; Polcar, T.; Gaudin, J.; Nagasono, M.; Sobierajski, R.; Yabashi, M.; Krzywinski, J.

    2013-01-01

    Roč. 21, č. 22 (2013), s. 26363-26375 ISSN 1094-4087 R&D Projects: GA ČR(CZ) GAP108/11/1312; GA ČR GA13-28721S; GA MŠk(CZ) LG13029; GA ČR GAP208/10/2302; GA ČR GAP205/11/0571; GA MŠk EE2.3.30.0057 Grant - others:AVČR(CZ) M100101221; OP VK 4 POSTDOK(XE) CZ.1.07/2.3.00/30.0057 Institutional support: RVO:68378271 Keywords : free-electron lasers (FELs) * UV * EUV * x-ray lasers * laser beam characterization * F-scan Subject RIV: BH - Optics, Masers, Lasers Impact factor: 3.525, year: 2013

  4. Laser Ultrasound Spectroscopy Scanning for 3D Printed Parts

    Energy Technology Data Exchange (ETDEWEB)

    Brennan, Guendalyn Kendra [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-08-04

    One of the challenges of additive manufacturing is quality control due to the possibility of unseen flaws in the final product. The current methods of inspection are lacking in detail, too slow for practical use, or unable to validate internal structure. This report examines the use of laser ultrasound spectroscopy in layer by layer scans of 3D printed parts as they are created. The result is fast and detailed quality control. An additional advantage of this method is the ability to cancel a print as soon as a defect is detected, therefore saving materials and time. This technique, though simple in concept, has been a challenge to implement. I discuss tweaking the 3D printer configuration, and finding the optimal settings for laser scanning small parts made of ABS plastic, as well as the limits of how small of a detail the laser can detect. These settings include the frequency of the ultrasonic transducer, the speed of the laser, and the distance from the laser to the part.

  5. Integrated Photoacoustic and Fluorescence Confocal Microscopy

    OpenAIRE

    Wang, Yu; Maslov, Konstantin; Kim, Chulhong; Hu, Song; Wang, Lihong V.

    2010-01-01

    We have developed a dual-modality imaging system by integrating optical-resolution photoacoustic microscopy and fluorescence confocal microscopy to provide optical absorption and fluorescence contrasts simultaneously. By sharing the same laser source and objective lens, intrinsically registered photoacoustic and fluorescence images are acquired in a single scan. The micrometer resolution allows imaging of both blood and lymphatic vessels down to the capillary level. Simultaneous photoacoustic...

  6. Laser scanning of a recirculation zone on the Bolund escarpment

    DEFF Research Database (Denmark)

    Mann, Jakob; Angelou, Nikolas; Sjöholm, Mikael

    2012-01-01

    Rapid variations in the height of the recirculation zone are measured with a scanning wind lidar over a small escarpment on the Bolund Peninsula. The lidar is essentially a continuous-wave laser Doppler anemometer with the capability of rapidly changing the focus distance and the beam direction....... The instrument measures the line-ofsight velocity 390 times per second and scans ten wind profiles from the ground up to seven meters per second. The results will be used to test computational fluid dynamics models for flow over terrain, and has relevance for wind energy. The development of multiple lidar...

  7. Laser scanning of a recirculation zone on the Bolund escarpment

    DEFF Research Database (Denmark)

    Mann, Jakob; Angelou, Nikolas; Sjöholm, Mikael

    2014-01-01

    Rapid variations in the height of the recirculation zone are measured with a scanning wind lidar over a small escarpment on the Bolund Peninsula. The lidar is essentially a continuous-wave laser Doppler anemometer with the capability of rapidly changing the focus distance and the beam direction....... The instrument measures the line-of-sight velocity 390 times per second and scans ten wind profiles from the ground up to seven meters per second. We observe a sharp interface between slow and fast moving fluid after the escarpment, and the interface is moving rapidly up and down. This implies that the position...

  8. Modeling 3D Objects for Navigation Purposes Using Laser Scanning

    Directory of Open Access Journals (Sweden)

    Cezary Specht

    2016-07-01

    Full Text Available The paper discusses the creation of 3d models and their applications in navigation. It contains a review of available methods and geometric data sources, focusing mostly on terrestrial laser scanning. It presents detailed description, from field survey to numerical elaboration, how to construct accurate model of a typical few storey building as a hypothetical reference in complex building navigation. Hence, the paper presents fields where 3d models are being used and their potential new applications.

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

    Science.gov (United States)

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

    Science.gov (United States)

    The confocal laser-scanning microscopy (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 the emitted signal. The accuracy of these measurements demands t...

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

    Science.gov (United States)

    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.

  12. Quality Assurance By Laser Scanning And Imaging Techniques

    Science.gov (United States)

    SchmalfuB, Harald J.; Schinner, Karl Ludwig

    1989-03-01

    Laser scanning systems are well established in the world of fast industrial in-process quality inspection systems. The materials inspected by laser scanning systems are e.g. "endless" sheets of steel, paper, textile, film or foils. The web width varies from 50 mm up to 5000 mm or more. The web speed depends strongly on the production process and can reach several hundred meters per minute. The continuous data flow in one of different channels of the optical receiving system exceeds ten Megapixels/sec. Therefore it is clear that the electronic evaluation system has to process these data streams in real time and no image storage is possible. But sometimes (e.g. first installation of the system, change of the defect classification) it would be very helpful to have the possibility for a visual look on the original, i.e. not processed sensor data. At first we show the principle set up of a standard laser scanning system. Then we will introduce a large image memory especially designed for the needs of high-speed inspection sensors. This image memory co-operates with the standard on-line evaluation electronics and provides therefore an easy comparison between processed and non-processed data. We will discuss the basic system structure and we will show the first industrial results.

  13. Calibration technology in application of robot-laser scanning system

    Science.gov (United States)

    Ren, YongJie; Yin, ShiBin; Zhu, JiGui

    2012-11-01

    A system composed of laser sensor and 6-DOF industrial robot is proposed to obtain complete three-dimensional (3-D) information of the object surface. Suitable for the different combining ways of laser sensor and robot, a new method to calibrate the position and pose between sensor and robot is presented. By using a standard sphere with known radius as a reference tool, the rotation and translation matrices between the laser sensor and robot are computed, respectively in two steps, so that many unstable factors introduced in conventional optimization methods can be avoided. The experimental results show that the accuracy of the proposed calibration method can be achieved up to 0.062 mm. The calibration method is also implemented into the automated robot scanning system to reconstruct a car door panel.

  14. 3-D laser confocal microscopy study of the oxidation of NdFeB magnets in atmospheric conditions

    International Nuclear Information System (INIS)

    Meakin, J.P.; Speight, J.D.; Sheridan, R.S.; Bradshaw, A.; Harris, I.R.; Williams, A.J.; Walton, A.

    2016-01-01

    Highlights: • Room temperature atmospheric oxidation behaviour of sintered NdFeB. • 3D laser confocal microscopy measurement of oxide phase growth. • Significant height increase of oxide phase only observed at triple points. • Raman spectroscopy identified oxide phase to be Nd 2 O 3 . • Diffusion coefficient determined to be 4 × 10 −13 cm 2 /s. - Abstract: Neodymium iron boron (NdFeB) magnets are used in a number of important applications, such as generators in gearless wind turbines, motors in electric vehicles and electronic goods (e.g.— computer hard disk drives, HDD). Hydrogen can be used as a processing gas to separate and recycle scrap sintered Nd-Fe-B magnets from end-of-life products to form a powder suitable for recycling. However, the magnets are likely to have been exposed to atmospheric conditions prior to processing, and any oxidation could lead to activation problems for the hydrogen decrepitation reaction. Many previous studies on the oxidation of NdFeB magnets have been performed at elevated temperatures; however, few studies have been formed under atmospheric conditions. In this paper a combination of 3-D laser confocal microscopy and Raman spectroscopy have been used to assess the composition, morphology and rate of oxidation/corrosion on scrap sintered NdFeB magnets. Confocal microscopy has been employed to measure the growth of surface reaction products at room temperature, immediately after exposure to air. The results showed that there was a significant height increase at the triple junctions of the Nd-rich grain boundaries. Using Raman spectroscopy, the product was shown to consist of Nd 2 O 3 and formed only on the Nd-rich triple junctions. The diffusion coefficient of the triple junction reaction product growth at 20 °C was determined to be approximately 4 × 10 −13 cm 2 /sec. This value is several orders of magnitude larger than values derived from the diffusion controlled oxide growth observations at elevated

  15. 3-D laser confocal microscopy study of the oxidation of NdFeB magnets in atmospheric conditions

    Energy Technology Data Exchange (ETDEWEB)

    Meakin, J.P., E-mail: jxm764@bham.ac.uk; Speight, J.D.; Sheridan, R.S.; Bradshaw, A.; Harris, I.R.; Williams, A.J.; Walton, A.

    2016-08-15

    Highlights: • Room temperature atmospheric oxidation behaviour of sintered NdFeB. • 3D laser confocal microscopy measurement of oxide phase growth. • Significant height increase of oxide phase only observed at triple points. • Raman spectroscopy identified oxide phase to be Nd{sub 2}O{sub 3}. • Diffusion coefficient determined to be 4 × 10{sup −13} cm{sup 2}/s. - Abstract: Neodymium iron boron (NdFeB) magnets are used in a number of important applications, such as generators in gearless wind turbines, motors in electric vehicles and electronic goods (e.g.— computer hard disk drives, HDD). Hydrogen can be used as a processing gas to separate and recycle scrap sintered Nd-Fe-B magnets from end-of-life products to form a powder suitable for recycling. However, the magnets are likely to have been exposed to atmospheric conditions prior to processing, and any oxidation could lead to activation problems for the hydrogen decrepitation reaction. Many previous studies on the oxidation of NdFeB magnets have been performed at elevated temperatures; however, few studies have been formed under atmospheric conditions. In this paper a combination of 3-D laser confocal microscopy and Raman spectroscopy have been used to assess the composition, morphology and rate of oxidation/corrosion on scrap sintered NdFeB magnets. Confocal microscopy has been employed to measure the growth of surface reaction products at room temperature, immediately after exposure to air. The results showed that there was a significant height increase at the triple junctions of the Nd-rich grain boundaries. Using Raman spectroscopy, the product was shown to consist of Nd{sub 2}O{sub 3} and formed only on the Nd-rich triple junctions. The diffusion coefficient of the triple junction reaction product growth at 20 °C was determined to be approximately 4 × 10{sup −13} cm{sup 2}/sec. This value is several orders of magnitude larger than values derived from the diffusion controlled oxide growth

  16. Improving Completeness of Geometric Models from Terrestrial Laser Scanning Data

    Directory of Open Access Journals (Sweden)

    Clemens Nothegger

    2011-12-01

    Full Text Available The application of terrestrial laser scanning for the documentation of cultural heritage assets is becoming increasingly common. While the point cloud by itself is sufficient for satisfying many documentation needs, it is often desirable to use this data for applications other than documentation. For these purposes a triangulated model is usually required. The generation of topologically correct triangulated models from terrestrial laser scans, however, still requires much interactive editing. This is especially true when reconstructing models from medium range panoramic scanners and many scan positions. Because of residual errors in the instrument calibration and the limited spatial resolution due to the laser footprint, the point clouds from different scan positions never match perfectly. Under these circumstances many of the software packages commonly used for generating triangulated models produce models which have topological errors such as surface intersecting triangles, holes or triangles which violate the manifold property. We present an algorithm which significantly reduces the number of topological errors in the models from such data. The algorithm is a modification of the Poisson surface reconstruction algorithm. Poisson surfaces are resilient to noise in the data and the algorithm always produces a closed manifold surface. Our modified algorithm partitions the data into tiles and can thus be easily parallelized. Furthermore, it avoids introducing topological errors in occluded areas, albeit at the cost of producing models which are no longer guaranteed to be closed. The algorithm is applied to scan data of sculptures of the UNESCO World Heritage Site Schönbrunn Palace and data of a petrified oyster reef in Stetten, Austria. The results of the method’s application are discussed and compared with those of alternative methods.

  17. Categorisation of full waveform data provided by laser scanning devices

    Science.gov (United States)

    Ullrich, Andreas; Pfennigbauer, Martin

    2011-11-01

    In 2004, a laser scanner device for commercial airborne laser scanning applications, the RIEGL LMS-Q560, was introduced to the market, making use of a radical alternative approach to the traditional analogue signal detection and processing schemes found in LIDAR instruments so far: digitizing the echo signals received by the instrument for every laser pulse and analysing these echo signals off-line in a so-called full waveform analysis in order to retrieve almost all information contained in the echo signal using transparent algorithms adaptable to specific applications. In the field of laser scanning the somewhat unspecific term "full waveform data" has since been established. We attempt a categorisation of the different types of the full waveform data found in the market. We discuss the challenges in echo digitization and waveform analysis from an instrument designer's point of view and we will address the benefits to be gained by using this technique, especially with respect to the so-called multi-target capability of pulsed time-of-flight LIDAR instruments.

  18. Measurement Axis Searching Model for Terrestrial Laser Scans Registration

    Directory of Open Access Journals (Sweden)

    Shaoxing Hu

    2016-01-01

    Full Text Available Nowadays, terrestrial Lidar scans can cover rather a large area; the point densities are strongly varied because of the line-of-sight measurement principle in potential overlaps with scans taken from different viewpoints. Most of the traditional methods focus on registration algorithm and ignore searching model. Sometimes the traditional methods are directly used to align two point clouds; a large critically unsolved problem of the large biases will be created in areas distant from the overlaps while the local overlaps are often aligned well. So a novel measurement axis searching model (MASM has been proposed in this paper. The method includes four steps: (1 the principal axis fitting, (2 the measurement axis generation, (3 low-high-precision search, and (4 result generation. The principal axis gives an orientation to the point cloud; the search scope is limited by the measurement axis. The point cloud orientation can be adjusted gradually until the achievement of the global optimum using low- and high-precision search. We perform some experiments with simulated point clouds and real terrestrial laser scans. The results of simulated point clouds have shown the processing steps of our method, and the results of real terrestrial laser scans have shown the sensitivity of the approach with respect to the indoor and outdoor scenes.

  19. Applications of confocal laser scanning microscopy in research into organic semiconductor thin films

    DEFF Research Database (Denmark)

    Schiek, Manuela; Balzer, Frank

    2014-01-01

    At the center of opto-electronic devices are thin layers of organic semiconductors, which need to be sandwiched between planar electrodes. With the growing demand for opto-electronic devices now and in the future, new electrode materials are needed to meet the requirements of organic semiconductors...

  20. Elastomeric photo-actuators and their investigation by confocal laser scanning microscopy

    Czech Academy of Sciences Publication Activity Database

    Czaniková, K.; Ilčíková, M.; Krupa, I.; Mičušík, M.; Kasák, P.; Pavlova, Ewa; Mosnáček, J.; Chorvát Jr., D.; Omastová, M.

    2013-01-01

    Roč. 22, č. 10 (2013), 104001_1-104001_10 ISSN 0964-1726 Institutional support: RVO:61389013 Keywords : nanocomposites * multiwalled carbon nanotubes * electron microscopy Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.449, year: 2013

  1. Fluorescence confocal laser scanning microscopy for in vivo imaging of epidermal reactions to two experimental irritants

    DEFF Research Database (Denmark)

    Suihko, C.; Serup, J.

    2008-01-01

    demonstrated the applicability of fluorescence CLSM for a detailed study of experimental skin irritants in vivo. Essential findings were disturbed and widened cell borders, swelling of keratinocytes by PA and induction of a parakeratotic shift by SLS with clusters of keratinocytes holding nuclei...... more complicated than reflectance CLSM and may not be applicable to any irritant. SLS applied epicutaneously interacted with the skin surface and coupling to the microscope and was thus found to be more difficult to study technically than PA. PA dissolved in isopropanol is for technical reasons...

  2. Laser cutting of irregular shape object based on stereo vision laser galvanometric scanning system

    Science.gov (United States)

    Qi, Li; Zhang, Yixin; Wang, Shun; Tang, Zhiqiang; Yang, Huan; Zhang, Xuping

    2015-05-01

    Irregular shape objects with different 3-dimensional (3D) appearances are difficult to be shaped into customized uniform pattern by current laser machining approaches. A laser galvanometric scanning system (LGS) could be a potential candidate since it can easily achieve path-adjustable laser shaping. However, without knowing the actual 3D topography of the object, the processing result may still suffer from 3D shape distortion. It is desirable to have a versatile auxiliary tool that is capable of generating 3D-adjusted laser processing path by measuring the 3D geometry of those irregular shape objects. This paper proposed the stereo vision laser galvanometric scanning system (SLGS), which takes the advantages of both the stereo vision solution and conventional LGS system. The 3D geometry of the object obtained by the stereo cameras is used to guide the scanning galvanometers for 3D-shape-adjusted laser processing. In order to achieve precise visual-servoed laser fabrication, these two independent components are integrated through a system calibration method using plastic thin film target. The flexibility of SLGS has been experimentally demonstrated by cutting duck feathers for badminton shuttle manufacture.

  3. Virtual pinhole confocal microscope

    Energy Technology Data Exchange (ETDEWEB)

    George, J.S.; Rector, D.M.; Ranken, D.M. [Los Alamos National Lab., NM (United States). Biophysics Group; Peterson, B. [SciLearn Inc. (United States); Kesteron, J. [VayTech Inc. (United States)

    1999-06-01

    Scanned confocal microscopes enhance imaging capabilities, providing improved contrast and image resolution in 3-D, but existing systems have significant technical shortcomings and are expensive. Researchers at Los Alamos National Laboratory have developed a novel approach--virtual pinhole confocal microscopy--that uses state of the art illumination, detection, and data processing technologies to produce an imager with a number of advantages: reduced cost, faster imaging, improved efficiency and sensitivity, improved reliability and much greater flexibility. Work at Los Alamos demonstrated proof of principle; prototype hardware and software have been used to demonstrate technical feasibility of several implementation strategies. The system uses high performance illumination, patterned in time and space. The authors have built functional confocal imagers using video display technologies (LCD or DLP) and novel scanner based on a micro-lens array. They have developed a prototype system for high performance data acquisition and processing, designed to support realtime confocal imaging. They have developed algorithms to reconstruct confocal images from a time series of spatially sub-sampled images; software development remains an area of active development. These advances allow the collection of high quality confocal images (in fluorescence, reflectance and transmission modes) with equipment that can inexpensively retrofit to existing microscopes. Planned future extensions to these technologies will significantly enhance capabilities for microscopic imaging in a variety of applications, including confocal endoscopy, and confocal spectral imaging.

  4. In-situ investigation of thermal instabilities and solid state dewetting in polycrystalline platinum thin films via confocal laser microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jahangir, S.; Cheng, Xuan; Huang, H. H.; Nagarajan, V. [School of Materials Science and Engineering, University of New South Wales, Sydney 2052 (Australia); Ihlefeld, J. [Electronic, Optical, and Nanomaterials Department, Sandia National Laboratories, Albuquerque, New Mexico 87185 (United States)

    2014-10-28

    Solid state dewetting and the subsequent morphological changes for platinum thin films grown on zinc oxide (ZnO) buffered (001) silicon substrates (Pt/ZnO/SiO{sub 2}/(001)Si system) is investigated under vacuum conditions via a custom-designed confocal laser microscope coupled with a laser heating system. Live imaging of thin film dewetting under a range of heating and quenching vacuum ambients reveals events including hillock formation, hole formation, and hole growth that lead to formation of a network of Pt ligaments, break up of Pt ligaments to individual islands and subsequent Pt islands shape reformation, in chronological fashion. These findings are corroborated by ex-situ materials characterization and quantitative electron microscopy analysis. A secondary hole formation via blistering before film rupture is revealed to be the critical stage, after which a rapid dewetting catastrophe occurs. This process is instantaneous and cannot be captured by ex-situ methods. Finally, an intermetallic phase forms at 900 °C and alters the morphology of Pt islands, suggesting a practical limit to the thermal environments that may be used for these platinized silicon wafers in vacuum conditions.

  5. In-situ investigation of thermal instabilities and solid state dewetting in polycrystalline platinum thin films via confocal laser microscopy

    International Nuclear Information System (INIS)

    Jahangir, S.; Cheng, Xuan; Huang, H. H.; Nagarajan, V.; Ihlefeld, J.

    2014-01-01

    Solid state dewetting and the subsequent morphological changes for platinum thin films grown on zinc oxide (ZnO) buffered (001) silicon substrates (Pt/ZnO/SiO 2 /(001)Si system) is investigated under vacuum conditions via a custom-designed confocal laser microscope coupled with a laser heating system. Live imaging of thin film dewetting under a range of heating and quenching vacuum ambients reveals events including hillock formation, hole formation, and hole growth that lead to formation of a network of Pt ligaments, break up of Pt ligaments to individual islands and subsequent Pt islands shape reformation, in chronological fashion. These findings are corroborated by ex-situ materials characterization and quantitative electron microscopy analysis. A secondary hole formation via blistering before film rupture is revealed to be the critical stage, after which a rapid dewetting catastrophe occurs. This process is instantaneous and cannot be captured by ex-situ methods. Finally, an intermetallic phase forms at 900 °C and alters the morphology of Pt islands, suggesting a practical limit to the thermal environments that may be used for these platinized silicon wafers in vacuum conditions.

  6. Laser sintering of metal powders on top of sintered layers under multiple-line laser scanning

    International Nuclear Information System (INIS)

    Xiao Bin; Zhang Yuwen

    2007-01-01

    A three-dimensional numerical model for multiple-line sintering of loose powders on top of multiple sintered layers under the irradiation of a moving Gaussian laser beam is carried out. The overlaps between vertically deposited layers and adjacent lines which strengthen bonding are taken into account. The energy equation is formulated using the temperature transforming model and solved by the finite volume method. The effects of the number of the existing sintered layers, porosity and initial temperature coupled with the optimal combination laser intensity and scanning velocity are presented. The results show that the liquid pool moves slightly towards the negative scanning direction and the shape of the liquid pool becomes shallower with higher scanning velocity. A higher laser intensity is needed to achieve the required overlaps when the number of the existing sintered layers increases. Increasing porosity or initial temperature enhances the sintering process and thus less intensity is needed for the overlap requirement

  7. Land-Based Mobile Laser Scanning Systems: a Review

    Science.gov (United States)

    Puente, I.; González-Jorge, H.; Arias, P.; Armesto, J.

    2011-09-01

    Mobile mapping has been using various photogrammetric techniques for many years. In recent years, there has been an increase in the number of mobile mapping systems using laser scanners available in the market, partially because of the improvement in GNSS/INS performance for direct georeferencing. In this article, some of the most important land-based mobile laser scanning (MLS) systems are reviewed. Firstly, the main characteristics of MLS systems vs. airborne (ALS) and terrestrial laser scanning (TLS) systems are compared. Secondly, a short overview of the mobile mapping technology is also provided so that the reader can fully grasp the complexity and operation of these devices. As we put forward in this paper, a comparison of different systems is briefly carried out regarding specifications provided by the manufacturers. Focuses on the current research are also addressed with emphasis on the practical applications of these systems. Most of them have been utilized for data collection on road infrastructures or building façades. This article shows that MLS technology is nowadays well established and proven, since the demand has grown to the point that there are several systems suppliers offering their products to satisfy this particular market.

  8. Codification of scan path parameters and development of perimeter scan strategies for 3D bowl-shaped laser forming

    Science.gov (United States)

    Tavakoli, A.; Naeini, H. Moslemi; Roohi, Amir H.; Gollo, M. Hoseinpour; Shahabad, Sh. Imani

    2018-01-01

    In the 3D laser forming process, developing an appropriate laser scan pattern for producing specimens with high quality and uniformity is critical. This study presents certain principles for developing scan paths. Seven scan path parameters are considered, including: (1) combined linear or curved path; (2) type of combined linear path; (3) order of scan sequences; (4) the position of the start point in each scan; (5) continuous or discontinuous scan path; (6) direction of scan path; and (7) angular arrangement of combined linear scan paths. Regarding these path parameters, ten combined linear scan patterns are presented. Numerical simulations show continuous hexagonal, scan pattern, scanning from outer to inner path, is the optimized. In addition, it is observed the position of the start point and the angular arrangement of scan paths is the most effective path parameters. Also, further experimentations show four sequences due to creat symmetric condition enhance the height of the bowl-shaped products and uniformity. Finally, the optimized hexagonal pattern was compared with the similar circular one. In the hexagonal scan path, distortion value and standard deviation rather to edge height of formed specimen is very low, and the edge height despite of decreasing length of scan path increases significantly compared to the circular scan path. As a result, four-sequence hexagonal scan pattern is proposed as the optimized perimeter scan path to produce bowl-shaped product.

  9. Deriving structural forest parameters using airborne laser scanning

    International Nuclear Information System (INIS)

    Morsdorf, F.

    2011-01-01

    Airborne laser scanning is a relatively young and precise technology to directly measure surface elevations. With today's high scanning rates, dense 3-D pointclouds of coordinate triplets (xyz) can be provided, in which many structural aspects of the vegetation are contained. The challenge now is to transform this data, as far as possible automatically, into manageable information relevant to the user. In this paper we present two such methods: the first extracts automatically the geometry of individual trees, with a recognition rate of over 70% and a systematic underestimation of tree height of only 0.6 metres. The second method derives a pixel map of the canopy density from the pointcloud, in which the spatial patterns of vegetation cover are represented. These patterns are relevant for habitat analysis and ecosystem studies. The values derived by this method correlate well with field measurements, giving a measure of certainty (R 2 ) of 0.8. The greatest advantage of airborne laser scanning is that it provides spatially extensive, direct measurements of vegetation structure which show none of the extrapolation errors of spot measurements. A large challenge remains in integrating these new products into the user's processing chains and workflows, be it in the realm of forestry or in that of ecosystem research. (author) [de

  10. Application of Laser Scanning for Creating Geological Documentation

    Directory of Open Access Journals (Sweden)

    Buczek Michał

    2018-01-01

    Full Text Available A geological documentation is based on the analyses obtained from boreholes, geological exposures, and geophysical methods. It consists of text and graphic documents, containing drilling sections, vertical crosssections through the deposit and various types of maps. The surveying methods (such as LIDAR can be applied in measurements of exposed rock layers, presented in appendices to the geological documentation. The laser scanning allows obtaining a complete profile of exposed surfaces in a short time and with a millimeter accuracy. The possibility of verifying the existing geological cross-section with laser scanning was tested on the example of the AGH experimental mine. The test field is built of different lithological rocks. Scans were taken from a single station, under favorable measuring conditions. The analysis of the signal intensity allowed to divide point cloud into separate geological layers. The results were compared with the geological profiles of the measured object. The same approach was applied to the data from the Vietnamese hard coal open pit mine Coc Sau. The thickness of exposed coal bed deposits and gangue layers were determined from the obtained data (point cloud in combination with the photographs. The results were compared with the geological cross-section.

  11. 3-D laser confocal microscopy study of the oxidation of NdFeB magnets in atmospheric conditions

    Science.gov (United States)

    Meakin, J. P.; Speight, J. D.; Sheridan, R. S.; Bradshaw, A.; Harris, I. R.; Williams, A. J.; Walton, A.

    2016-08-01

    Neodymium iron boron (NdFeB) magnets are used in a number of important applications, such as generators in gearless wind turbines, motors in electric vehicles and electronic goods (e.g.- computer hard disk drives, HDD). Hydrogen can be used as a processing gas to separate and recycle scrap sintered Nd-Fe-B magnets from end-of-life products to form a powder suitable for recycling. However, the magnets are likely to have been exposed to atmospheric conditions prior to processing, and any oxidation could lead to activation problems for the hydrogen decrepitation reaction. Many previous studies on the oxidation of NdFeB magnets have been performed at elevated temperatures; however, few studies have been formed under atmospheric conditions. In this paper a combination of 3-D laser confocal microscopy and Raman spectroscopy have been used to assess the composition, morphology and rate of oxidation/corrosion on scrap sintered NdFeB magnets. Confocal microscopy has been employed to measure the growth of surface reaction products at room temperature, immediately after exposure to air. The results showed that there was a significant height increase at the triple junctions of the Nd-rich grain boundaries. Using Raman spectroscopy, the product was shown to consist of Nd2O3 and formed only on the Nd-rich triple junctions. The diffusion coefficient of the triple junction reaction product growth at 20 °C was determined to be approximately 4 × 10-13 cm2/sec. This value is several orders of magnitude larger than values derived from the diffusion controlled oxide growth observations at elevated temperatures in the literature. This indicates that the growth of the room temperature oxidation products are likely defect enhanced processes at the NdFeB triple junctions.

  12. Scanning laser beam displays based on a 2D MEMS

    Science.gov (United States)

    Niesten, Maarten; Masood, Taha; Miller, Josh; Tauscher, Jason

    2010-05-01

    The combination of laser light sources and MEMS technology enables a range of display systems such as ultra small projectors for mobile devices, head-up displays for vehicles, wearable near-eye displays and projection systems for 3D imaging. Images are created by scanning red, green and blue lasers horizontally and vertically with a single two-dimensional MEMS. Due to the excellent beam quality of laser beams, the optical designs are efficient and compact. In addition, the laser illumination enables saturated display colors that are desirable for augmented reality applications where a virtual image is used. With this technology, the smallest projector engine for high volume manufacturing to date has been developed. This projector module has a height of 7 mm and a volume of 5 cc. The resolution of this projector is WVGA. No additional projection optics is required, resulting in an infinite focus depth. Unlike with micro-display projection displays, an increase in resolution will not lead to an increase in size or a decrease in efficiency. Therefore future projectors can be developed that combine a higher resolution in an even smaller and thinner form factor with increased efficiencies that will lead to lower power consumption.

  13. AUTOMATIC RAILWAY POWER LINE EXTRACTION USING MOBILE LASER SCANNING DATA

    Directory of Open Access Journals (Sweden)

    S. Zhang

    2016-06-01

    Full Text Available Research on power line extraction technology using mobile laser point clouds has important practical significance on railway power lines patrol work. In this paper, we presents a new method for automatic extracting railway power line from MLS (Mobile Laser Scanning data. Firstly, according to the spatial structure characteristics of power-line and trajectory, the significant data is segmented piecewise. Then, use the self-adaptive space region growing method to extract power lines parallel with rails. Finally use PCA (Principal Components Analysis combine with information entropy theory method to judge a section of the power line whether is junction or not and which type of junction it belongs to. The least squares fitting algorithm is introduced to model the power line. An evaluation of the proposed method over a complicated railway point clouds acquired by a RIEGL VMX450 MLS system shows that the proposed method is promising.

  14. Resolution Enhancement of Scanning Laser Acoustic Microscope Using Transverse Wave

    International Nuclear Information System (INIS)

    Ko, D. S.; Park, J. S.; Kim, Y. H.

    1997-01-01

    We studied the resolution enhancement of a novel scanning laser acoustic microscope (SLAM) using transverse waves. Mode conversion of the ultrasonic wave takes place at the liquid-solid interface and some energy of the insonifying longitudinal waves in the water will convert to transverse wave energy within the solid specimen. The resolution of SLAM depends on the size of detecting laser spot and the wavelength of the insonifying ultrasonic waves. Science the wavelength of the transverse wave is shorter than that of the longitudinal wave, we are able to achieve the high resolution by using transverse waves. In order to operate SLAM in the transverse wave mode, we made wedge for changing the incident angle. Our experimental results with model 2140 SLAM and an aluminum specimen showed higher contrast of the SLAM image in the transverse wave mode than that in the longitudinal wave mode

  15. 2-photon laser scanning microscopy on native human cartilage

    Science.gov (United States)

    Martini, Joerg; Toensing, Katja; Dickob, Michael; Anselmetti, Dario

    2005-08-01

    Native hyaline cartilage from a human knee joint was directly investigated with laser scanning microscopy via 2-photon autofluorescence excitation with no additional staining or labelling protocols in a nondestructive and sterile manner. Using a femtosecond, near-infrared (NIR) Ti:Sa laser for 2-photon excitation and a dedicated NIR long distance objective, autofluorescence imaging and measurements of the extracellular matrix (ECM) tissue with incorporated chondrocytes were possible with a penetration depth of up to 460 μm inside the sample. Via spectral autofluorescence separation these experiments allowed the discrimination of chondrocytes from the ECM and therefore an estimate of chondrocytic cell density within the cartilage tissue to approximately 0.2-2•107cm3. Furthermore, a comparison of the relative autofluorescence signals between nonarthritic and arthritic cartilage tissue exhibited distinct differences in tissue morphology. As these morphological findings are in keeping with the macroscopic diagnosis, our measurement has the potential of being used in future diagnostic applications.

  16. Surface characterization of weathered wood using a laser scanning system

    International Nuclear Information System (INIS)

    Arnold, M.; Lemaster, R.L.; Dost, W.A.

    1992-01-01

    Most of the existing methods to assess the effect of weathering on wood surfaces have some drawbacks that limit their use to specific tasks. The amount of surface erosion is often used as a measure for the weathering action. The application of a laser scanning system to reproduce surface profiles and to measure weathering erosion was tested on various samples and was found to be a very useful and superior alternative to existing methods. Further improvements of the system used can be made by refinements of the calibration procedures and by more comprehensive profile analyses. (author)

  17. Cryogenic scanning laser microscopy. Investigation of large BSCCO mesas and development of a polarizing microscope

    International Nuclear Information System (INIS)

    Guenon, Stefan Alexander

    2011-01-01

    confirmed that the frequency of the emitted radiation and the bias voltage is determined by the Josephson relation for a wide range of different base temperatures. This way other mechanisms, causing THz radiation, rather than the Josephson effect can be excluded. Concerning the second part: Originally it was planned to extend the low-temperature scanning laser microscope with the facility of polarizing microscopy. The idea was to combine the LTSLM voltage imaging with the possibility of magneto-optical imaging. But it soon turned out that a new design would be necessary. A laser scanning polarizing microscope has certain advantages in comparison with a conventional polarizing microscope: Very high illumination intensities can be reached easily, the resolution can be improved by the factor 1.4 if a confocal optical design is used, and the serial signal processing facilitates the optimization of the signal-to-noise ratio. In addition, it is usually not necessary to remove the contrast of non-magnetic origin by subtracting an image of the uniform magnetized sample from the image of interest. In this thesis a design for a cryogenic scanning polarizing microscope (CSPM) is discussed in detail, tests and first results of the system are presented, and an outlook is given how two proceed with this project.

  18. Determination of foveal location using scanning laser polarimetry.

    Science.gov (United States)

    VanNasdale, Dean A; Elsner, Ann E; Weber, Anke; Miura, Masahiro; Haggerty, Bryan P

    2009-03-25

    The fovea is the retinal location responsible for our most acute vision. There are several methods used to localize the fovea, but the fovea is not always easily identifiable. Landmarks used to determine the foveal location are variable in normal subjects and localization becomes even more difficult in instances of retinal disease. In normal subjects, the photoreceptor axons that make up the Henle fiber layer are cylindrical and the radial orientation of these fibers is centered on the fovea. The Henle fiber layer exhibits form birefringence, which predictably changes polarized light in scanning laser polarimetry imaging. In this study 3 graders were able to repeatably identify the fovea in 35 normal subjects using near infrared image types with differing polarization content. There was little intra-grader, inter-grader, and inter-image variability in the graded foveal position for 5 of the 6 image types examined, with accuracy sufficient for clinical purposes. This study demonstrates that scanning laser polarimetry imaging can localize the fovea by using structural properties inherent in the central macula.

  19. Automatic Indoor Building Reconstruction from Mobile Laser Scanning Data

    Science.gov (United States)

    Xie, L.; Wang, R.

    2017-09-01

    Indoor reconstruction from point clouds is a hot topic in photogrammetry, computer vision and computer graphics. Reconstructing indoor scene from point clouds is challenging due to complex room floorplan and line-of-sight occlusions. Most of existing methods deal with stationary terrestrial laser scanning point clouds or RGB-D point clouds. In this paper, we propose an automatic method for reconstructing indoor 3D building models from mobile laser scanning point clouds. The method includes 2D floorplan generation, 3D building modeling, door detection and room segmentation. The main idea behind our approach is to separate wall structure into two different types as the inner wall and the outer wall based on the observation of point distribution. Then we utilize a graph cut based optimization method to solve the labeling problem and generate the 2D floorplan based on the optimization result. Subsequently, we leverage an ?-shape based method to detect the doors on the 2D projected point clouds and utilize the floorplan to segment the individual room. The experiments show that this door detection method can achieve a recognition rate at 97% and the room segmentation method can attain the correct segmentation results. We also evaluate the reconstruction accuracy on the synthetic data, which indicates the accuracy of our method is comparable to the state-of-the art.

  20. Adaptive optics scanning laser ophthalmoscope imaging: technology update

    Directory of Open Access Journals (Sweden)

    Merino D

    2016-04-01

    Full Text Available David Merino, Pablo Loza-Alvarez The Institute of Photonic Sciences (ICFO, The Barcelona Institute of Science and Technology, Castelldefels, Barcelona, Spain Abstract: Adaptive optics (AO retinal imaging has become very popular in the past few years, especially within the ophthalmic research community. Several different retinal techniques, such as fundus imaging cameras or optical coherence tomography systems, have been coupled with AO in order to produce impressive images showing individual cell mosaics over different layers of the in vivo human retina. The combination of AO with scanning laser ophthalmoscopy has been extensively used to generate impressive images of the human retina with unprecedented resolution, showing individual photoreceptor cells, retinal pigment epithelium cells, as well as microscopic capillary vessels, or the nerve fiber layer. Over the past few years, the technique has evolved to develop several different applications not only in the clinic but also in different animal models, thanks to technological developments in the field. These developments have specific applications to different fields of investigation, which are not limited to the study of retinal diseases but also to the understanding of the retinal function and vision science. This review is an attempt to summarize these developments in an understandable and brief manner in order to guide the reader into the possibilities that AO scanning laser ophthalmoscopy offers, as well as its limitations, which should be taken into account when planning on using it. Keywords: high-resolution, in vivo retinal imaging, AOSLO

  1. Application of terrestrial laser scanning for measuring tree crown structures

    International Nuclear Information System (INIS)

    Pretzsch, H.; Seifert, S.; Huang, P.

    2011-01-01

    This paper addresses the potential of terrestrial laser scanning (TLS) for describing and modelling of tree crown structure and dynamics. We first present a general approach for the metabolic and structural scaling of tree crowns. Out of this approach we emphasize those normalization and scaling parameters which become accessible by TLS. For example we show how the individual tree leaf area index, convex hull, and its space-filling by leaves can be extracted out of laser scan data. This contributes to a theoretical and empirical substantiation of crown structure models which were missing so far for e.g. quantification of structural and species diversity in forest stands, inventory of crown biomass, species detection by remote sensing, and understanding of self- and alien-thinning in pure and mixed stands. Up to now works on this topic delivered a rather scattered empirical knowledge mainly by single inventories of trees and stands. In contrast, we recommend to start with a model approach, and to complete existing data with repeated TLS inventories in order to come to a consistent and theoretically based model of tree crowns. (author) [de

  2. Confocal Adaptive Optics Imaging of Peripapillary Nerve Fiber Bundles: Implications for Glaucomatous Damage Seen on Circumpapillary OCT Scans.

    Science.gov (United States)

    Hood, Donald C; Chen, Monica F; Lee, Dongwon; Epstein, Benjamin; Alhadeff, Paula; Rosen, Richard B; Ritch, Robert; Dubra, Alfredo; Chui, Toco Y P

    2015-04-01

    To improve our understanding of glaucomatous damage as seen on circumpapillary disc scans obtained with frequency-domain optical coherence tomography (fdOCT), fdOCT scans were compared to images of the peripapillary retinal nerve fiber (RNF) bundles obtained with an adaptive optics-scanning light ophthalmoscope (AO-SLO). The AO-SLO images and fdOCT scans were obtained on 6 eyes of 6 patients with deep arcuate defects (5 points ≤-15 db) on 10-2 visual fields. The AO-SLO images were montaged and aligned with the fdOCT images to compare the RNF bundles seen with AO-SLO to the RNF layer thickness measured with fdOCT. All 6 eyes had an abnormally thin (1% confidence limit) RNF layer (RNFL) on fdOCT and abnormal (hyporeflective) regions of RNF bundles on AO-SLO in corresponding regions. However, regions of abnormal, but equal, RNFL thickness on fdOCT scans varied in appearance on AO-SLO images. These regions could be largely devoid of RNF bundles (5 eyes), have abnormal-appearing bundles of lower contrast (6 eyes), or have isolated areas with a few relatively normal-appearing bundles (2 eyes). There also were local variations in reflectivity of the fdOCT RNFL that corresponded to the variations in AO-SLO RNF bundle appearance. Relatively similar 10-2 defects with similar fdOCT RNFL thickness profiles can have very different degrees of RNF bundle damage as seen on fdOCT and AO-SLO. While the results point to limitations of fdOCT RNFL thickness as typically analyzed, they also illustrate the potential for improving fdOCT by attending to variations in local intensity.

  3. Test field for airborne laser scanning in Finland

    Science.gov (United States)

    Ahokas, E.; Kaartinen, H.; Kukko, A.; Litkey, P.

    2014-11-01

    Airborne laser scanning (ALS) is a widely spread operational measurement tool for obtaining 3D coordinates of the ground surface. There is a need for calibrating the ALS system and a test field for ALS was established at the end of 2013. The test field is situated in the city of Lahti, about 100 km to the north of Helsinki. The size of the area is approximately 3.5 km × 3.2 km. Reference data was collected with a mobile laser scanning (MLS) system assembled on a car roof. Some streets were measured both ways and most of them in one driving direction only. The MLS system of the Finnish Geodetic Institute (FGI) consists of a navigation system (NovAtel SPAN GNSS-IMU) and a laser scanner (FARO Focus3D 120). In addition to the MLS measurements more than 800 reference points were measured using a Trimble R8 VRS-GNSS system. Reference points are along the streets, on parking lots, and white pedestrian crossing line corners which can be used as reference targets. The National Land Survey of Finland has already used this test field this spring for calibrating their Leica ALS-70 scanner. Especially it was easier to determine the encoder scale factor parameter using this test field. Accuracy analysis of the MLS points showed that the point height RMSE is 2.8 cm and standard deviation is 2.6 cm. Our purpose is to measure both more MLS data and more reference points in the test field area to get a better spatial coverage. Calibration flight heights are planned to be 1000 m and 2500 m above ground level. A cross pattern, southwest-northeast and northwest-southeast, will be flown both in opposite directions.

  4. Flexible Spectral Imaging Color Enhancement and Probe-based Confocal Laser Endomicroscopy in Minimal Change Esophageal Reflux Disease.

    Science.gov (United States)

    Pittayanon, Rapat; Aumkaew, Surasak; Rerknimitr, Rungsun; Wisedopas, Naruemon; Kullavanijaya, Pinit

    2016-07-25

    Although flexible spectral imaging color enhancement (FICE) can facilitate the diagnosis of minimal change esophageal reflux disease (MERD), the complicated diagnostic criteria cause suboptimal inter-observer agreement. Confocal laser endomicroscopy (CLE) yields good diagnostic results but its inter-observer agreement has never been explored. This study compares the diagnostic value of magnifying FICE and probe-based CLE (pCLE) for MERD and evaluates the inter-observer agreement of both techniques. Thirty-six patients with suspected MERD and 18 asymptomatic controls were recruited. Magnifying FICE was used for evaluation of distal esophagus. pCLE counted the number of intrapapillary capillary loops (IPCLs) using more than five IPCLs in 500×500 micron area as a criterion for MERD diagnosis. The validity scores and interobserever agreement of both FICE and pCLE were assessed. For FICE vs. pCLE, the accuracy was 79% vs. 87%, sensitivity 94% vs. 97%, specificity 50% vs. 66%, positive predictive value 79% vs. 85%, and negative predictive value 82% vs. 92%. Interobserver agreement of FICE was fair to substantial, whereas pCLE had substantial to almost perfect agreement. Both FICE and pCLE have good operating characteristics and can facilitate the MERD diagnosis. However, among different observers, pCLE is more consistent on MERD diagnosis.

  5. Confocal comparison of corneal reinnervation after small incision lenticule extraction (SMILE) and femtosecond laser in situ keratomileusis (FS-LASIK).

    Science.gov (United States)

    Li, Meiyan; Niu, Lingling; Qin, Bing; Zhou, Zimei; Ni, Katherine; Le, Qihua; Xiang, Jun; Wei, Anji; Ma, Weiping; Zhou, Xingtao

    2013-01-01

    To evaluate corneal reinnervation, and the corresponding corneal sensitivity and keratocyte density after small incision lenticule extraction (SMILE) and femtosecond laser in situ keratomileusis (FS-LASIK). In this prospective, non-randomized observational study, 18 patients (32 eyes) received SMILE surgery, and 22 patients (42 eyes) received FS-LASIK surgery to correct myopia. The corneal subbasal nerve density and microscopic morphological changes in corneal architecture were evaluated by confocal microscopy prior to surgery and at 1 week, 1 month, 3 months, and 6 months after surgery. A correlation analysis was performed between subbasal corneal nerve density and the corresponding keratocyte density and corneal sensitivity. The decrease in subbasal nerve density was less severe in SMILE-treated eyes than in FS-LASIK-treated eyes at 1 week (P = 0.0147), 1 month (P = 0.0243), and 3 months (P = 0.0498), but no difference was detected at the 6-month visit (P = 0.5277). The subbasal nerve density correlated positively with central corneal sensitivity in both groups (r = 0.416, PLASIK group, respectively). The SMILE-treated eyes have a lower risk of developing peripheral empty space with epithelial cells filling in (P = 0.0005). The decrease in subbasal nerve fiber density was less severe in the SMILE group than the FS-LASIK group in the first 3 months following the surgeries. The subbasal nerve density was correlated with central corneal sensitivity.

  6. Expert-led didactic versus self-directed audiovisual training of confocal laser endomicroscopy in evaluation of mucosal barrier defects.

    Science.gov (United States)

    Huynh, Roy; Ip, Matthew; Chang, Jeff; Haifer, Craig; Leong, Rupert W

    2018-01-01

    Confocal laser endomicroscopy (CLE) allows mucosal barrier defects along the intestinal epithelium to be visualized in vivo during endoscopy. Training in CLE interpretation can be achieved didactically or through self-directed learning. This study aimed to compare the effectiveness of expert-led didactic with self-directed audiovisual teaching for training inexperienced analysts on how to recognize mucosal barrier defects on endoscope-based CLE (eCLE).  This randomized controlled study involved trainee analysts who were taught how to recognize mucosal barrier defects on eCLE either didactically or through an audiovisual clip. After being trained, they evaluated 6 sets of 30 images. Image evaluation required the trainees to determine whether specific features of barrier dysfunction were present or not. Trainees in the didactic group engaged in peer discussion and received feedback after each set while this did not happen in the self-directed group. Accuracy, sensitivity, and specificity of both groups were compared. Trainees in the didactic group achieved a higher overall accuracy (87.5 % vs 85.0 %, P  = 0.002) and sensitivity (84.5 % vs 80.4 %, P  = 0.002) compared to trainees in the self-directed group. Interobserver agreement was higher in the didactic group (k = 0.686, 95 % CI 0.680 - 0.691, P  barrier defects on eCLE.

  7. Multicolor probe-based confocal laser endomicroscopy: a new world for in vivo and real-time cellular imaging

    Science.gov (United States)

    Vercauteren, Tom; Doussoux, François; Cazaux, Matthieu; Schmid, Guillaume; Linard, Nicolas; Durin, Marie-Amélie; Gharbi, Hédi; Lacombe, François

    2013-03-01

    Since its inception in the field of in vivo imaging, endomicroscopy through optical fiber bundles, or probe-based Confocal Laser Endomicroscopy (pCLE), has extensively proven the benefit of in situ and real-time examination of living tissues at the microscopic scale. By continuously increasing image quality, reducing invasiveness and improving system ergonomics, Mauna Kea Technologies has turned pCLE not only into an irreplaceable research instrument for small animal imaging, but also into an accurate clinical decision making tool with applications as diverse as gastrointestinal endoscopy, pulmonology and urology. The current implementation of pCLE relies on a single fluorescence spectral band making different sources of in vivo information challenging to distinguish. Extending the pCLE approach to multi-color endomicroscopy therefore appears as a natural plan. Coupling simultaneous multi-laser excitation with minimally invasive, microscopic resolution, thin and flexible optics, allows the fusion of complementary and valuable biological information, thus paving the way to a combination of morphological and functional imaging. This paper will detail the architecture of a new system, Cellvizio Dual Band, capable of video rate in vivo and in situ multi-spectral fluorescence imaging with a microscopic resolution. In its standard configuration, the system simultaneously operates at 488 and 660 nm, where it automatically performs the necessary spectral, photometric and geometric calibrations to provide unambiguously co-registered images in real-time. The main hardware and software features, including calibration procedures and sub-micron registration algorithms, will be presented as well as a panorama of its current applications, illustrated with recent results in the field of pre-clinical imaging.

  8. Pavement cracking measurements using 3D laser-scan images

    International Nuclear Information System (INIS)

    Ouyang, W; Xu, B

    2013-01-01

    Pavement condition surveying is vital for pavement maintenance programs that ensure ride quality and traffic safety. This paper first introduces an automated pavement inspection system which uses a three-dimensional (3D) camera and a structured laser light to acquire dense transverse profiles of a pavement lane surface when it carries a moving vehicle. After the calibration, the 3D system can yield a depth resolution of 0.5 mm and a transverse resolution of 1.56 mm pixel −1 at 1.4 m camera height from the ground. The scanning rate of the camera can be set to its maximum at 5000 lines s −1 , allowing the density of scanned profiles to vary with the vehicle's speed. The paper then illustrates the algorithms that utilize 3D information to detect pavement distress, such as transverse, longitudinal and alligator cracking, and presents the field tests on the system's repeatability when scanning a sample pavement in multiple runs at the same vehicle speed, at different vehicle speeds and under different weather conditions. The results show that this dedicated 3D system can capture accurate pavement images that detail surface distress, and obtain consistent crack measurements in repeated tests and under different driving and lighting conditions. (paper)

  9. Research on calibration algorithm in laser scanning projection system

    Science.gov (United States)

    Li, Li Juan; Qu, Song; Hou, Mao Sheng

    2017-10-01

    Laser scanning projection technology can project the image defined by the existing CAD digital model to the working surface, in the form of a laser harness profile. This projection is in accordance with the ratio of 1: 1. Through the laser harness contours with high positioning quality, the technical staff can carry out the operation with high precision. In a typical process of the projection, in order to determine the relative positional relationship between the laser projection instrument and the target, it is necessary to place several fixed reference points on the projection target and perform the calibration of projection. This position relationship is the transformation from projection coordinate system to the global coordinate system. The entire projection work is divided into two steps: the first step, the calculation of the projector six position parameters is performed, that is, the projector calibration. In the second step, the deflection angle is calculated by the known projector position parameter and the known coordinate points, and then the actual model is projected. Typically, the calibration requires the establishment of six reference points to reduce the possibility of divergence of the nonlinear equations, but the whole solution is very complex and the solution may still diverge. In this paper, the distance is detected combined with the calculation so that the position parameters of the projector can be solved by using the coordinate values of three reference points and the distance of at least one reference point to the projector. The addition of the distance measurement increases the stability of the solution of the nonlinear system and avoids the problem of divergence of the solution caused by the reference point which is directly under the projector. Through the actual analysis and calculation, the Taylor expansion method combined with the least squares method is used to obtain the solution of the system. Finally, the simulation experiment is

  10. Confocal laser endomicroscopy and ultrasound endoscopy during the same endoscopic session for diagnosis and staging of gastric neoplastic lesions.

    Science.gov (United States)

    Gheorghe, C; Iacob, R; Dumbrava, Mona; Becheanu, G; Ionescu, M

    2009-01-01

    Confocal LASER endomicroscopy (CLE) is a newly developed endoscopic technique which allows subsurface in vivo histological assessment during ongoing endoscopy and targeted biopsies. Ultrasound endoscopy (EUS) is a useful tool in staging upper GI malignant lesions. We describe for the first time the use of both techniques during the same endoscopic session, in a pilot study, in order to increase the diagnostic yield of histological assessment and provide the staging of the gastric neoplastic lesions thus decreasing the time to therapeutic decision. CLE has been performed with the Pentax EG-3870CIK confocal endomicroscope after a 5 ml intravenous 10% fluorescein injection; EUS has been performed subsequently, during the same endoscopic Propofol sedation session, using a standard radial EUS-scope. Eleven patients have been investigated, 4 females, 7 males, mean age 59.7 +/- 12.3 years. The indication of CLE/EUS exploration was the presence of a gastric polypoid lesion in 37% of cases, atypical gastric ulcer in 27% of patients, gastric lymphoma 18%, suspicion of gastric cancer recurrence after resection 9% and infiltrating type gastric cancer 9%. Histological assessment after targeted biopsy has established the diagnosis of gastric adenocarcinoma in 55% of cases, gastric lymphoma in 18% of cases, gastric adenoma, gastric GIST and gastric foveolar hyperplasia in 9% of cases respectively. CLE has allowed targeted biopsies in 81.8% of cases. In 2 patients - one case with suspected recurrent gastric cancer after surgery and one case of gastric lymphoma, CLE has indicated normal gastric mucosa. The EUS evaluation has shown TO lesion in two cases, T1 in 3 cases, T2 in 3 cases, T3 in one case. The EUS evaluation showed in one gastric lymphoma patient a lesion interesting the mucosa and submucosa with regional adenopathy and a submucosal lesion with regional adenopathy in the other gastric lymphoma case. The therapeutic decision was surgery in 73% of cases, chemotherapy and

  11. Terrestrial Laser Scanning-Based Bridge Structural Condition Assessment : Tech Transfer Summaries

    Science.gov (United States)

    2016-05-01

    Problem Statement : While several state departments of transportation (DOTs) have used : terrestrial laser scanning (TLS) in the project planning phase, limited : research has been conducted on employing laser scanners to detect : cracks for bridge c...

  12. Spinning-disk confocal microscopy: present technology and future trends.

    Science.gov (United States)

    Oreopoulos, John; Berman, Richard; Browne, Mark

    2014-01-01

    Live-cell imaging requires not only high temporal resolution but also illumination powers low enough to minimize photodamage. Traditional single-point laser scanning confocal microscopy (LSCM) is generally limited by both the relatively slow speed at which it can acquire optical sections by serial raster scanning (a few Hz) and the higher potential for phototoxicity. These limitations have driven the development of rapid, parallel forms of confocal microscopy, the most popular of which is the spinning-disk confocal microscope (SDCM). Here, we briefly introduce the SDCM technique, discuss its strengths and weaknesses against LSCM, and update the reader on some recent developments in SDCM technology that improve its performance and expand its utility for life science research now and in the future. © 2014 Elsevier Inc. All rights reserved.

  13. Classification of Mobile Laser Scanning Point Clouds from Height Features

    Science.gov (United States)

    Zheng, M.; Lemmens, M.; van Oosterom, P.

    2017-09-01

    The demand for 3D maps of cities and road networks is steadily growing and mobile laser scanning (MLS) systems are often the preferred geo-data acquisition method for capturing such scenes. Because MLS systems are mounted on cars or vans they can acquire billions of points of road scenes within a few hours of survey. Manual processing of point clouds is labour intensive and thus time consuming and expensive. Hence, the need for rapid and automated methods for 3D mapping of dense point clouds is growing exponentially. The last five years the research on automated 3D mapping of MLS data has tremendously intensified. In this paper, we present our work on automated classification of MLS point clouds. In the present stage of the research we exploited three features - two height components and one reflectance value, and achieved an overall accuracy of 73 %, which is really encouraging for further refining our approach.

  14. 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.

    Science.gov (United States)

    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.

  15. Monitoring of Progressive Damage in Buildings Using Laser Scan Data

    Science.gov (United States)

    Puente, I.; Lindenbergh, R.; Van Natijne, A.; Esposito, R.; Schipper, R.

    2018-05-01

    Vulnerability of buildings to natural and man-induced hazards has become a main concern for our society. Ensuring their serviceability, safety and sustainability is of vital importance and the main reason for setting up monitoring systems to detect damages at an early stage. In this work, a method is presented for detecting changes from laser scan data, where no registration between different epochs is needed. To show the potential of the method, a case study of a laboratory test carried out at the Stevin laboratory of Delft University of Technology was selected. The case study was a quasi-static cyclic pushover test on a two-story high unreinforced masonry structure designed to simulate damage evolution caused by cyclic loading. During the various phases, we analysed the behaviour of the masonry walls by monitoring the deformation of each masonry unit. First a plane is fitted to the selected wall point cloud, consisting of one single terrestrial laser scan, using Principal Component Analysis (PCA). Second, the segmentation of individual elements is performed. Then deformations with respect to this plane model, for each epoch and specific element, are determined by computing their corresponding rotation and cloud-to-plane distances. The validation of the changes detected within this approach is done by comparison with traditional deformation analysis based on co-registered TLS point clouds between two or more epochs of building measurements. Initial results show that the sketched methodology is indeed able to detect changes at the mm level while avoiding 3D point cloud registration, which is a main issue in computer vision and remote sensing.

  16. AUTOMATIC EXTRACTION OF ROAD MARKINGS FROM MOBILE LASER SCANNING DATA

    Directory of Open Access Journals (Sweden)

    H. Ma

    2017-09-01

    Full Text Available Road markings as critical feature in high-defination maps, which are Advanced Driver Assistance System (ADAS and self-driving technology required, have important functions in providing guidance and information to moving cars. Mobile laser scanning (MLS system is an effective way to obtain the 3D information of the road surface, including road markings, at highway speeds and at less than traditional survey costs. This paper presents a novel method to automatically extract road markings from MLS point clouds. Ground points are first filtered from raw input point clouds using neighborhood elevation consistency method. The basic assumption of the method is that the road surface is smooth. Points with small elevation-difference between neighborhood are considered to be ground points. Then ground points are partitioned into a set of profiles according to trajectory data. The intensity histogram of points in each profile is generated to find intensity jumps in certain threshold which inversely to laser distance. The separated points are used as seed points to region grow based on intensity so as to obtain road mark of integrity. We use the point cloud template-matching method to refine the road marking candidates via removing the noise clusters with low correlation coefficient. During experiment with a MLS point set of about 2 kilometres in a city center, our method provides a promising solution to the road markings extraction from MLS data.

  17. Automatic Extraction of Road Markings from Mobile Laser Scanning Data

    Science.gov (United States)

    Ma, H.; Pei, Z.; Wei, Z.; Zhong, R.

    2017-09-01

    Road markings as critical feature in high-defination maps, which are Advanced Driver Assistance System (ADAS) and self-driving technology required, have important functions in providing guidance and information to moving cars. Mobile laser scanning (MLS) system is an effective way to obtain the 3D information of the road surface, including road markings, at highway speeds and at less than traditional survey costs. This paper presents a novel method to automatically extract road markings from MLS point clouds. Ground points are first filtered from raw input point clouds using neighborhood elevation consistency method. The basic assumption of the method is that the road surface is smooth. Points with small elevation-difference between neighborhood are considered to be ground points. Then ground points are partitioned into a set of profiles according to trajectory data. The intensity histogram of points in each profile is generated to find intensity jumps in certain threshold which inversely to laser distance. The separated points are used as seed points to region grow based on intensity so as to obtain road mark of integrity. We use the point cloud template-matching method to refine the road marking candidates via removing the noise clusters with low correlation coefficient. During experiment with a MLS point set of about 2 kilometres in a city center, our method provides a promising solution to the road markings extraction from MLS data.

  18. PEDESTRIAN DETECTION BY LASER SCANNING AND DEPTH IMAGERY

    Directory of Open Access Journals (Sweden)

    A. Barsi

    2016-06-01

    Full Text Available Pedestrian flow is much less regulated and controlled compared to vehicle traffic. Estimating flow parameters would support many safety, security or commercial applications. Current paper discusses a method that enables acquiring information on pedestrian movements without disturbing and changing their motion. Profile laser scanner and depth camera have been applied to capture the geometry of the moving people as time series. Procedures have been developed to derive complex flow parameters, such as count, volume, walking direction and velocity from laser scanned point clouds. Since no images are captured from the faces of pedestrians, no privacy issues raised. The paper includes accuracy analysis of the estimated parameters based on video footage as reference. Due to the dense point clouds, detailed geometry analysis has been conducted to obtain the height and shoulder width of pedestrians and to detect whether luggage has been carried or not. The derived parameters support safety (e.g. detecting critical pedestrian density in mass events, security (e.g. detecting prohibited baggage in endangered areas and commercial applications (e.g. counting pedestrians at all entrances/exits of a shopping mall.

  19. Pedestrian Detection by Laser Scanning and Depth Imagery

    Science.gov (United States)

    Barsi, A.; Lovas, T.; Molnar, B.; Somogyi, A.; Igazvolgyi, Z.

    2016-06-01

    Pedestrian flow is much less regulated and controlled compared to vehicle traffic. Estimating flow parameters would support many safety, security or commercial applications. Current paper discusses a method that enables acquiring information on pedestrian movements without disturbing and changing their motion. Profile laser scanner and depth camera have been applied to capture the geometry of the moving people as time series. Procedures have been developed to derive complex flow parameters, such as count, volume, walking direction and velocity from laser scanned point clouds. Since no images are captured from the faces of pedestrians, no privacy issues raised. The paper includes accuracy analysis of the estimated parameters based on video footage as reference. Due to the dense point clouds, detailed geometry analysis has been conducted to obtain the height and shoulder width of pedestrians and to detect whether luggage has been carried or not. The derived parameters support safety (e.g. detecting critical pedestrian density in mass events), security (e.g. detecting prohibited baggage in endangered areas) and commercial applications (e.g. counting pedestrians at all entrances/exits of a shopping mall).

  20. Confocal comparison of corneal reinnervation after small incision lenticule extraction (SMILE and femtosecond laser in situ keratomileusis (FS-LASIK.

    Directory of Open Access Journals (Sweden)

    Meiyan Li

    Full Text Available PURPOSE: To evaluate corneal reinnervation, and the corresponding corneal sensitivity and keratocyte density after small incision lenticule extraction (SMILE and femtosecond laser in situ keratomileusis (FS-LASIK. METHODS: In this prospective, non-randomized observational study, 18 patients (32 eyes received SMILE surgery, and 22 patients (42 eyes received FS-LASIK surgery to correct myopia. The corneal subbasal nerve density and microscopic morphological changes in corneal architecture were evaluated by confocal microscopy prior to surgery and at 1 week, 1 month, 3 months, and 6 months after surgery. A correlation analysis was performed between subbasal corneal nerve density and the corresponding keratocyte density and corneal sensitivity. RESULTS: The decrease in subbasal nerve density was less severe in SMILE-treated eyes than in FS-LASIK-treated eyes at 1 week (P = 0.0147, 1 month (P = 0.0243, and 3 months (P = 0.0498, but no difference was detected at the 6-month visit (P = 0.5277. The subbasal nerve density correlated positively with central corneal sensitivity in both groups (r = 0.416, P<0.0001, and r = 0.2567, P = 0.0038 for SMILE group and FS-LASIK group, respectively. The SMILE-treated eyes have a lower risk of developing peripheral empty space with epithelial cells filling in (P = 0.0005. CONCLUSIONS: The decrease in subbasal nerve fiber density was less severe in the SMILE group than the FS-LASIK group in the first 3 months following the surgeries. The subbasal nerve density was correlated with central corneal sensitivity.

  1. Evaluation of the therapeutic results of actinic keratosis treated with topical 5% fluorouracil by reflectance confocal laser microscopy: preliminary study*

    Science.gov (United States)

    Ishioka, Priscila; Maia, Marcus; Rodrigues, Sarita Bartholomei; Marta, Alessandra Cristina; Hirata, Sérgio Henrique

    2015-01-01

    Topical treatment for actinic keratosis with 5% fluorouracil has a recurrence rate of 54% in 12 months of follow-up. This study analyzed thirteen actinic keratoses on the upper limbs through confocal microscopy, at the time of clinical diagnosis and after 4 weeks of treatment with fluorouracil. After the treatment was established and evidence of clinical cure was achieved, in two of the nine actinic keratoses, confocal microscopy enabled visualization of focal areas of atypical honeycomb pattern in the epidermis indicating therapeutic failure. Preliminary data suggest the use of confocal microscopy as a tool for diagnosis and therapeutic control of actinic keratosis. PMID:26131881

  2. Long-term efficacy of linear-scanning 808 nm diode laser for hair removal compared to a scanned alexandrite laser.

    Science.gov (United States)

    Grunewald, Sonja; Bodendorf, Marc Oliver; Zygouris, Alexander; Simon, Jan Christoph; Paasch, Uwe

    2014-01-01

    Alexandrite and diode lasers are commonly used for hair removal. To date, the available spot sizes and repetition rates are defining factors in terms of penetration depth, treatment speed, and efficacy. Still, larger treatment areas and faster systems are desirable. To compare the efficacy, tolerability, and subject satisfaction of a continuously linear-scanning 808 nm diode laser with an alexandrite 755 nm laser for axillary hair removal. A total of 31 adults with skin types I-IV received 6 treatments at 4-week intervals with a 755 nm alexandrite laser (right axilla) and a continuously linear-scanning 808 nm diode laser (left axilla). Axillary hair density was assessed using a computerized hair detection system. There was a significant reduction in axillary hair after the 6th treatment (P lasers was not significant, but both were persistant at 18 months follow-up (left: hair clearance of 73.71%; right: hair clearance of 71.90%). Erythema and perifollicular edema were more common after alexandrite laser treatment, but all side effects were transient. While 62.50% of patients reported more pain in response to treatment with the new diode laser, all patients rated treatment with either laser tolerable. Treatment with either the alexandrite or the linear-scanning diode laser results in significant, comparable, persistent (at least 18 months) axillary hair reduction among individuals with skin types I-IV. © 2013 Wiley Periodicals, Inc.

  3. Deformation Monitoring of Motorway Underpasses Using Laser Scanning Data

    Science.gov (United States)

    Puente, I.; González-Jorge, H.; Riveiro, B.; Arias, P.

    2012-07-01

    is a Optech Lynx mobile LiDAR. This laser scanner is based on time of flight technology and presents an accuracy of 6 mm in the determination of the geometrical coordinates. This accuracy can be improved to around 1 mm using fitting post-processing techniques and makes this technology very useful for studies related with deformation monitoring. The laser scanner, in comparison with other geodetic techniques as total stations, allows the control of all the structure, including unexpected deformations. Reflective targets are permanently positioned over the small walls of the structure to allow the 3D orientation of the different scans. Two main scans are made for this study, before and after the backfilling process. Backfilling takes about 10 days for the construction companies. The scans need a time of approximately 12 minutes. Construction works do not need to be interrupted during the scans. Point clouds are then post-processed using QT Modeler Software. First, the point cloud is cleaned to use only the data directly related with the structure under study. Then, using the target coordinates, both point clouds are moved to the same coordinate system. Finally, the deformation of the underpass is studied using two algorithms specifically developed using Matlab software. First algorithm fits a geometrical surface to the point cloud of the first scan and evaluates the residuals of both scans for this fitting surface. Differences in the residuals give the deformation map of the structure. Second algorithm takes a portion of the point cloud from the top of the structure, where it is located the joining point between the voussoirs. The joining between two voussoirs shows a height step that in an ideal case must tend to zero. Deformations produced by the loading of the structure are measured as a comparison between the steps before and after the backfilling process. The analysis of the results show as some deformation occurs in the structure in the joining point of the

  4. DEFORMATION MONITORING OF MOTORWAY UNDERPASSES USING LASER SCANNING DATA

    Directory of Open Access Journals (Sweden)

    I. Puente

    2012-07-01

    deformation monitoring is a Optech Lynx mobile LiDAR. This laser scanner is based on time of flight technology and presents an accuracy of 6 mm in the determination of the geometrical coordinates. This accuracy can be improved to around 1 mm using fitting post-processing techniques and makes this technology very useful for studies related with deformation monitoring. The laser scanner, in comparison with other geodetic techniques as total stations, allows the control of all the structure, including unexpected deformations. Reflective targets are permanently positioned over the small walls of the structure to allow the 3D orientation of the different scans. Two main scans are made for this study, before and after the backfilling process. Backfilling takes about 10 days for the construction companies. The scans need a time of approximately 12 minutes. Construction works do not need to be interrupted during the scans. Point clouds are then post-processed using QT Modeler Software. First, the point cloud is cleaned to use only the data directly related with the structure under study. Then, using the target coordinates, both point clouds are moved to the same coordinate system. Finally, the deformation of the underpass is studied using two algorithms specifically developed using Matlab software. First algorithm fits a geometrical surface to the point cloud of the first scan and evaluates the residuals of both scans for this fitting surface. Differences in the residuals give the deformation map of the structure. Second algorithm takes a portion of the point cloud from the top of the structure, where it is located the joining point between the voussoirs. The joining between two voussoirs shows a height step that in an ideal case must tend to zero. Deformations produced by the loading of the structure are measured as a comparison between the steps before and after the backfilling process. The analysis of the results show as some deformation occurs in the structure in the joining

  5. Scanning laser topography and scanning laser polarimetry: comparing both imaging methods at same distances from the optic nerve head.

    Science.gov (United States)

    Kremmer, Stephan; Keienburg, Marcus; Anastassiou, Gerasimos; Schallenberg, Maurice; Steuhl, Klaus-Peter; Selbach, J Michael

    2012-01-01

    To compare the performance of scanning laser topography (SLT) and scanning laser polarimetry (SLP) on the rim of the optic nerve head and its surrounding area and thereby to evaluate whether these imaging technologies are influenced by other factors beyond the thickness of the retinal nerve fiber layer (RNFL). A total of 154 eyes from 5 different groups were examined: young healthy subjects (YNorm), old healthy subjects (ONorm), patients with normal tension glaucoma (NTG), patients with open-angle glaucoma and early glaucomatous damage (OAGE) and patients with open-angle glaucoma and advanced glaucomatous damage (OAGA). SLT and SLP measurements were taken. Four concentric circles were superimposed on each of the images: the first one measuring at the rim of the optic nerve head (1.0 ONHD), the next measuring at 1.25 optic nerve head diameters (ONHD), at 1.5 ONHD and at 1.75 ONHD. The aligned images were analyzed using GDx/NFA software. Both methods showed peaks of RNFL thickness in the superior and inferior segments of the ONH. The maximum thickness, registered by the SLT device was at the ONH rim where the SLP device tended to measure the lowest values. SLT measurements at the ONH were influenced by other tissues besides the RNFL like blood vessels and glial tissues. SLT and SLP were most strongly correlated at distances of 1.25 and 1.5 ONHD. While both imaging technologies are valuable tools in detecting glaucoma, measurements at the ONH rim should be interpreted critically since both methods might provide misleading results. For the assessment of the retinal nerve fiber layer we would like to recommend for both imaging technologies, SLT and SLP, measurements in 1.25 and 1.5 ONHD distance of the rim of the optic nerve head.

  6. Confocal Imaging of porous media

    Science.gov (United States)

    Shah, S.; Crawshaw, D.; Boek, D.

    2012-12-01

    Carbonate rocks, which hold approximately 50% of the world's oil and gas reserves, have a very complicated and heterogeneous structure in comparison with sandstone reservoir rock. We present advances with different techniques to image, reconstruct, and characterize statistically the micro-geometry of carbonate pores. The main goal here is to develop a technique to obtain two dimensional and three dimensional images using Confocal Laser Scanning Microscopy. CLSM is used in epi-fluorescent imaging mode, allowing for the very high optical resolution of features well below 1μm size. Images of pore structures were captured using CLSM imaging where spaces in the carbonate samples were impregnated with a fluorescent, dyed epoxy-resin, and scanned in the x-y plane by a laser probe. We discuss the sample preparation in detail for Confocal Imaging to obtain sub-micron resolution images of heterogeneous carbonate rocks. We also discuss the technical and practical aspects of this imaging technique, including its advantages and limitation. We present several examples of this application, including studying pore geometry in carbonates, characterizing sub-resolution porosity in two dimensional images. We then describe approaches to extract statistical information about porosity using image processing and spatial correlation function. We have managed to obtain very low depth information in z -axis (~ 50μm) to develop three dimensional images of carbonate rocks with the current capabilities and limitation of CLSM technique. Hence, we have planned a novel technique to obtain higher depth information to obtain high three dimensional images with sub-micron resolution possible in the lateral and axial planes.

  7. Effects of 915 nm GaAs diode laser on mitochondria of human dermal fibroblasts: analysis with confocal microscopy.

    Science.gov (United States)

    Belletti, Silvana; Uggeri, Jacopo; Mergoni, Giovanni; Vescovi, Paolo; Merigo, Elisabetta; Fornaini, Carlo; Nammour, Samir; Manfredi, Maddalena; Gatti, Rita

    2015-01-01

    Low-level laser therapy (LLLT) is widely used in tissue regeneration and pain therapy. Mitochondria are supposed to be one of the main cellular targets, due to the presence of cytochrome C oxidase as photo-acceptor. Laser stimulation could influence mitochondria metabolism affecting mainly transmembrane mitochondrial potential (Δψm). The aim of our study is to evaluate "in vitro" the early mitochondrial response after irradiation with a 915 GaAs laser. Since some evidences suggest that cellular response to LLLT can be differently modulated by the mode of irradiation, we would like to evaluate whether there are changes in the mitochondrial potential linked to the use of the laser treatments applied with continuous wave (CW) in respect to those applied with pulsed wave (PW). In this study, we analyzed effects of irradiation with a 915-nm GaAs diode laser on human dermal fibroblast. We compared effects of irradiation applied with either CW or PW at different fluences 45-15-5 J/cm(2) on Δψm. Laser scanning microscopy (LSM) was used in living cells to detect ROS (reactive oxygen species) using calcein AM and real-time changes of and Δψm following distribution of the potentiometric probe tetramethylrhodamine methyl ester (TMRM). At higher doses (45-15 J/cm(2)), fibroblasts showed a dose-dependent decrement of Δψm in either the modalities employed, with higher amplitudes in CW-treated cells. This behavior is transient and not followed by any sign of toxicity, even if reactive oxygen species generation was observed. At 5 J/cm(2), CW irradiation determined a little decrease (5%) of the baseline level of Δψm, while opposite behavior was shown when cells were irradiated with PW, with a 10% increment. Our results suggest that different responses observed at cellular level with low doses of irradiation, could be at the basis of efficacy of LLLT in clinical application, performed with PW rather than CW modalities.

  8. New insights into the painting stratigraphy of L'Homme blesse by Gustave Courbet combining scanning macro-XRF and confocal micro-XRF

    International Nuclear Information System (INIS)

    Reiche, Ina; Eveno, Myriam; Pichon, Laurent; Laval, Eric; Mottin, Bruno; Mueller, Katharina; Calligaro, Thomas; Mysak, Erin

    2016-01-01

    The painting L'Homme blesse by Gustave Courbet kept at the Musee d'Orsay in Paris has been recently studied by X-ray radiography, SEM-EDX observation of paint cross sections and confocal micro-X-ray fluorescence analyses (CXRF) at locations where the cross section samples were taken. This study allowed the establishment of the paint palette used by Courbet for the three paint compositions. Eight or more paint layers could be evidenced. In the view of the complexity of this painting, further analyses using two-dimensional scanning macro-X-ray fluorescence imaging (MA-XRF) providing chemical images corresponding to the superimposition of all detectable paint layers were employed. This method is combined with CXRF for depth-resolved paint layer analysis. Large elemental maps of Hg, Cu, As, Fe, Zn, Cr, Ba, Pb and Ca were obtained by MA-XRF on the painting and are discussed in combination with depth profiles obtained by CXRF on strategic points where three painting compositions overlap. The order of three successive compositions of this painting were determined in this study. This work also highlights the benefits of using complementary imaging methods to obtain a complete three-dimensional vision of the chemistry and stratigraphy of paintings. (orig.)

  9. New insights into the painting stratigraphy of L'Homme blesse by Gustave Courbet combining scanning macro-XRF and confocal micro-XRF

    Energy Technology Data Exchange (ETDEWEB)

    Reiche, Ina [Staatliche Museen zu Berlin-Preussischer Kulturbesitz, Rathgen-Forschungslabor, Berlin (Germany); Laboratoire d' Archeologie Moleculaire et Structurale, Sorbonne Universites, Univ. Paris 06, CNRS, UMR 8220, Paris (France); Eveno, Myriam; Pichon, Laurent; Laval, Eric; Mottin, Bruno [Centre de Recherche et de Restauration des Musees de France (C2RMF), Paris (France); Mueller, Katharina [Laboratoire d' Archeologie Moleculaire et Structurale, Sorbonne Universites, Univ. Paris 06, CNRS, UMR 8220, Paris (France); Calligaro, Thomas [Centre de Recherche et de Restauration des Musees de France (C2RMF), Paris (France); PSL Research University, Chimie ParisTech-CNRS, Institut de Recherche Chimie Paris, Paris (France); Mysak, Erin [Centre de Recherche et de Restauration des Musees de France (C2RMF), Paris (France); Yale University, Institute for the Preservation of Cultural Heritage, New Haven, CT (United States)

    2016-11-15

    The painting L'Homme blesse by Gustave Courbet kept at the Musee d'Orsay in Paris has been recently studied by X-ray radiography, SEM-EDX observation of paint cross sections and confocal micro-X-ray fluorescence analyses (CXRF) at locations where the cross section samples were taken. This study allowed the establishment of the paint palette used by Courbet for the three paint compositions. Eight or more paint layers could be evidenced. In the view of the complexity of this painting, further analyses using two-dimensional scanning macro-X-ray fluorescence imaging (MA-XRF) providing chemical images corresponding to the superimposition of all detectable paint layers were employed. This method is combined with CXRF for depth-resolved paint layer analysis. Large elemental maps of Hg, Cu, As, Fe, Zn, Cr, Ba, Pb and Ca were obtained by MA-XRF on the painting and are discussed in combination with depth profiles obtained by CXRF on strategic points where three painting compositions overlap. The order of three successive compositions of this painting were determined in this study. This work also highlights the benefits of using complementary imaging methods to obtain a complete three-dimensional vision of the chemistry and stratigraphy of paintings. (orig.)

  10. Macrophages and dendritic cells in the rat meninges and choroid plexus: three-dimensional localisation by environmental scanning electron microscopy and confocal microscopy.

    Science.gov (United States)

    McMenamin, Paul G; Wealthall, Rosamund J; Deverall, Marie; Cooper, Stephanie J; Griffin, Brendan

    2003-09-01

    The present investigation provides novel information on the topographical distribution of macrophages and dendritic cells (DCs) in normal meninges and choroid plexus of the rat central nervous system (CNS). Whole-mounts of meninges and choroid plexus of Lewis rats were incubated with various anti-leucocyte monoclonal antibodies and either visualised with gold-conjugated secondary antibody followed by silver enhancement and subsequent examination by environmental scanning electron microscopy or by the use of fluorochromes and confocal microscopy. Large numbers of MHC class II(+) putative DCs were identified on the internal or subarachnoid aspect of dural whole-mounts, on the surface of the cortex (pia/arachnoid) and on the surface of the choroid plexus. Occupation of these sites would allow DCs access to cerebrospinal fluid (CSF) and therefore allow antigens into the subarachnoid space and ventricles. By contrast, macrophages were less evident at sites exposed to CSF and were more frequently located within the connective tissue of the dura/arachnoid and choroid plexus stroma and also in a sub-pial location. The present data suggest that DC may be strategically located within the CNS to sample CSF-borne antigens. Furthermore, the data suggest that CNS tissue samples collected without careful removal of the meninges may inadvertently be contaminated by DCs and meningeal macrophages.

  11. Characterization of atopic skin and the effect of a hyperforin-rich cream by laser scanning microscopy

    Science.gov (United States)

    Meinke, Martina C.; Richter, Heike; Kleemann, Anke; Lademann, Juergen; Tscherch, Kathrin; Rohn, Sascha; Schempp, Christoph M.

    2015-05-01

    Atopic dermatitis (AD) is a multifactorial inflammatory skin disease that affects both children and adults in an increasing manner. The treatment of AD often reduces subjective skin parameters, such as itching, dryness, and tension, but the inflammation cannot be cured. Laser scanning microscopy was used to investigate the skin surface, epidermal, and dermal characteristics of dry and atopic skin before and after treatment with an ointment rich in hyperforin, which is known for its anti-inflammatory effects. The results were compared to subjective parameters and transepidermal water loss, stratum corneum moisture, and stratum corneum lipids. Using biophysical methods, in particular laser scanning microscopy, it was found that atopic skin has distinct features compared to healthy skin. Treatment with a hyperforin-rich ointment resulted in an improvement of the stratum corneum moisture, skin surface dryness, skin lipids, and the subjective skin parameters, indicating that the barrier is stabilized and improved by the ointment. But in contrast to the improved skin surface, the inflammation in the deeper epidermis/dermis often continues to exist. This could be clearly shown by the reflectance confocal microscopy (RCM) measurements. Therefore, RCM measurements could be used to investigate the progress in treatment of atopic dermatitis.

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

    Science.gov (United States)

    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.

  13. Surveying a fossil oyster reef using terrestrial laser scanning

    Science.gov (United States)

    Haring, A.; Exner, U.; Harzhauser, M.

    2009-04-01

    The Korneuburg Basin, situated north-west of Vienna, is well known to contain a rich variety of fossils from the Early Miocene (16.5 ma) and therefore has been investigated extensively by scientists in the past decades. An exceptional discovery was made in 2005: a large fossil oyster reef has been excavated and documented carefully during the last years. Aside from the giant-sized oyster (Crassostrea gryphoides), the excavation site contains numerous species of molluscs along with teeth of sharks and rays and even isolated bones of sea cows. The oysters, having lengths of up to 80 cm, are protruding from the ground surface, which is more or less a tilted plane (25˚ ) with a size of about 300 m2. The entire site is crosscut by a network of geological faults, often also offsetting individual oyster shells. Displacements along the normal faults do not exceed ~ 15 cm. The faulted fossils offer a unique opportunity to measure displacement distribution along the faults in great detail and provide insight in deformation mechanisms in porous, barely lithified sediments. In order to get a precise 3D model of the oyster reef, the terrestrial laser scanner system Leica HDS 6000 is used. It is a phase-based laser scanner, i.e. the distance measurement is performed using the phase-shift principle. Compared to the time-of-flight principle, this method is generally more appropriate to projects like this one, where the distances to be measured are relatively small (< 35 m) and where a high point density (point spacing of about 1 cm) and precision (some mm) is required for capturing the oysters adequately. However, due to fact that they occlude each other, one single scan is not sufficient to get all sides of their surface. Therefore, scans from different positions had to be acquired. These scans have to be merged, which involves the problem of sensor orientation as well as sampling of the entire 3D point cloud. Furthermore, a representation of the surface data is required that

  14. Retinal nerve fiber layer assessment by scanning laser polarimetry and standardized photography

    NARCIS (Netherlands)

    Niessen, A. G.; van den Berg, T. J.; Langerhorst, C. T.; Greve, E. L.

    1996-01-01

    To determine whether, in a clinical setting, scanning laser polarimetry and retinal nerve fiber layer photography provide equivalent information on the retinal nerve fiber layer. We prospectively studied 60 patients with glaucoma or ocular hypertension and 24 healthy subjects. With scanning laser

  15. Estimation of forest resources from a country wide laser scanning survey and national forest inventory data

    DEFF Research Database (Denmark)

    Nord-Larsen, Thomas; Schumacher, Johannes

    2012-01-01

    Airborne laser scanning may provide a means for assessing local forest biomass resources. In this study, national forest inventory (NFI) data was used as reference data for modeling forest basal area, volume, aboveground biomass, and total biomass from laser scanning data obtained in a countrywid...

  16. Tree Classification with Fused Mobile Laser Scanning and Hyperspectral Data

    Science.gov (United States)

    Puttonen, Eetu; Jaakkola, Anttoni; Litkey, Paula; Hyyppä, Juha

    2011-01-01

    Mobile Laser Scanning data were collected simultaneously with hyperspectral data using the Finnish Geodetic Institute Sensei system. The data were tested for tree species classification. The test area was an urban garden in the City of Espoo, Finland. Point clouds representing 168 individual tree specimens of 23 tree species were determined manually. The classification of the trees was done using first only the spatial data from point clouds, then with only the spectral data obtained with a spectrometer, and finally with the combined spatial and hyperspectral data from both sensors. Two classification tests were performed: the separation of coniferous and deciduous trees, and the identification of individual tree species. All determined tree specimens were used in distinguishing coniferous and deciduous trees. A subset of 133 trees and 10 tree species was used in the tree species classification. The best classification results for the fused data were 95.8% for the separation of the coniferous and deciduous classes. The best overall tree species classification succeeded with 83.5% accuracy for the best tested fused data feature combination. The respective results for paired structural features derived from the laser point cloud were 90.5% for the separation of the coniferous and deciduous classes and 65.4% for the species classification. Classification accuracies with paired hyperspectral reflectance value data were 90.5% for the separation of coniferous and deciduous classes and 62.4% for different species. The results are among the first of their kind and they show that mobile collected fused data outperformed single-sensor data in both classification tests and by a significant margin. PMID:22163894

  17. Pattern scan laser versus single spot laser in panretinal photocoagulation treatment for proliferative diabetic retinopathy

    Directory of Open Access Journals (Sweden)

    Shu Zhang

    2017-02-01

    Full Text Available AIM: To investigate the efficacy of 577-nm pattern scan laser in panretinal photocoagulation(PRPtreatment in newly diagnosed proliferative diabetic retinopathy(PDR.METHODS:Prospective and comparative observation was performed in totally 32 patients with high-risk PDR. They were randomly divided into group 1(using pattern scan laser, PSLand 2(using single spot laser, SSL, each containing 16 subjects to which totally 20 eyes received PRP. Non-perfusion region was identified with fundus fluorescein angiography(FFAbefore and 3mo after final PRP. The advantage of PSL was verified in terms of the number and the duration of PRP sessions needed for satisfactory outcomes, and the pain score.RESULTS: Three PRP sessions were needed for each eye to complete the treatment using PSL, while 4 sessions were needed using SSL. The duration of each session with PSL in group 1 was 7.3±2.3min, which was significantly shorter than that with SSL in group 2(13.2±4.1, t38=5.596, PPCONCLUSION: PSL showed clear advantages over SSL in the PRP treatment of PDR, not only in the improved efficacy, but also in the reduction of pain and the improvement of effectiveness.

  18. Laser scanning dental probe for endodontic root canal treatment

    Science.gov (United States)

    Blank, Molly A. B.; Friedrich, Michal; Hamilton, Jeffrey D.; Lee, Peggy; Berg, Joel; Seibel, Eric J.

    2011-03-01

    Complications that arise during endodontic procedures pose serious threats to the long-term integrity and health of the tooth. Potential complexities of root canals include residual pulpal tissue, cracks, mesial-buccal 2 and accessory canals. In the case of a failed root canal, a successful apicoectomy can be jeopardized by isthmuses, accessory canals, and root microfracture. Confirming diagnosis using a small imaging probe would allow proper treatment and prevent retreatment of endodontic procedures. An ultrathin and flexible laser scanning endoscope of 1.2 to 1.6mm outer diameter was used in vitro to image extracted teeth with varied root configurations. Teeth were opened using a conventional bur and high speed drill. Imaging within the opened access cavity clarified the location of the roots where canal filing would initiate. Although radiographs are commonly used to determine the root canal size, position, and shape, the limited 2D image perspective leaves ambiguity that could be clarified if used in conjunction with a direct visual imaging tool. Direct visualization may avoid difficulties in locating the root canal and reduce the number of radiographs needed. A transillumination imaging device with the separated illumination and light collection functions rendered cracks visible in the prepared teeth that were otherwise indiscernible using reflected visible light. Our work demonstrates that a small diameter endoscope with high spatial resolution may significantly increase the efficiency and success of endodontic procedures.

  19. AN AUTOMATED ROAD ROUGHNESS DETECTION FROM MOBILE LASER SCANNING DATA

    Directory of Open Access Journals (Sweden)

    P. Kumar

    2017-05-01

    Full Text Available Rough roads influence the safety of the road users as accident rate increases with increasing unevenness of the road surface. Road roughness regions are required to be efficiently detected and located in order to ensure their maintenance. Mobile Laser Scanning (MLS systems provide a rapid and cost-effective alternative by providing accurate and dense point cloud data along route corridor. In this paper, an automated algorithm is presented for detecting road roughness from MLS data. The presented algorithm is based on interpolating smooth intensity raster surface from LiDAR point cloud data using point thinning process. The interpolated surface is further processed using morphological and multi-level Otsu thresholding operations to identify candidate road roughness regions. The candidate regions are finally filtered based on spatial density and standard deviation of elevation criteria to detect the roughness along the road surface. The test results of road roughness detection algorithm on two road sections are presented. The developed approach can be used to provide comprehensive information to road authorities in order to schedule maintenance and ensure maximum safety conditions for road users.

  20. An Automated Road Roughness Detection from Mobile Laser Scanning Data

    Science.gov (United States)

    Kumar, P.; Angelats, E.

    2017-05-01

    Rough roads influence the safety of the road users as accident rate increases with increasing unevenness of the road surface. Road roughness regions are required to be efficiently detected and located in order to ensure their maintenance. Mobile Laser Scanning (MLS) systems provide a rapid and cost-effective alternative by providing accurate and dense point cloud data along route corridor. In this paper, an automated algorithm is presented for detecting road roughness from MLS data. The presented algorithm is based on interpolating smooth intensity raster surface from LiDAR point cloud data using point thinning process. The interpolated surface is further processed using morphological and multi-level Otsu thresholding operations to identify candidate road roughness regions. The candidate regions are finally filtered based on spatial density and standard deviation of elevation criteria to detect the roughness along the road surface. The test results of road roughness detection algorithm on two road sections are presented. The developed approach can be used to provide comprehensive information to road authorities in order to schedule maintenance and ensure maximum safety conditions for road users.

  1. Measuring Leaf Water Content Using Multispectral Terrestrial Laser Scanning

    Science.gov (United States)

    Junttila, S.; Vastaranta, M.; Linnakoski, R.; Sugano, J.; Kaartinen, H.; Kukko, A.; Holopainen, M.; Hyyppä, H.; Hyyppä, J.

    2017-10-01

    Climate change is increasing the amount and intensity of disturbance events, i.e. drought, pest insect outbreaks and fungal pathogens, in forests worldwide. Leaf water content (LWC) is an early indicator of tree stress that can be measured remotely using multispectral terrestrial laser scanning (MS-TLS). LWC affects leaf reflectance in the shortwave infrared spectrum which can be used to predict LWC from spatially explicit MS-TLS intensity data. Here, we investigated the relationship between LWC and MS-TLS intensity features at 690 nm, 905 nm and 1550 nm wavelengths with Norway spruce seedlings in greenhouse conditions. We found that a simple ratio of 905 nm and 1550 nm wavelengths was able to explain 84 % of the variation (R2) in LWC with a respective prediction accuracy of 0.0041 g/cm2. Our results showed that MS-TLS can be used to estimate LWC with a reasonable accuracy in environmentally stable conditions.

  2. Hierarchical extraction of urban objects from mobile laser scanning data

    Science.gov (United States)

    Yang, Bisheng; Dong, Zhen; Zhao, Gang; Dai, Wenxia

    2015-01-01

    Point clouds collected in urban scenes contain a huge number of points (e.g., billions), numerous objects with significant size variability, complex and incomplete structures, and variable point densities, raising great challenges for the automated extraction of urban objects in the field of photogrammetry, computer vision, and robotics. This paper addresses these challenges by proposing an automated method to extract urban objects robustly and efficiently. The proposed method generates multi-scale supervoxels from 3D point clouds using the point attributes (e.g., colors, intensities) and spatial distances between points, and then segments the supervoxels rather than individual points by combining graph based segmentation with multiple cues (e.g., principal direction, colors) of the supervoxels. The proposed method defines a set of rules for merging segments into meaningful units according to types of urban objects and forms the semantic knowledge of urban objects for the classification of objects. Finally, the proposed method extracts and classifies urban objects in a hierarchical order ranked by the saliency of the segments. Experiments show that the proposed method is efficient and robust for extracting buildings, streetlamps, trees, telegraph poles, traffic signs, cars, and enclosures from mobile laser scanning (MLS) point clouds, with an overall accuracy of 92.3%.

  3. Monitoring Riverbank Erosion in Mountain Catchments Using Terrestrial Laser Scanning

    Directory of Open Access Journals (Sweden)

    Laura Longoni

    2016-03-01

    Full Text Available Sediment yield is a key factor in river basins management due to the various and adverse consequences that erosion and sediment transport in rivers may have on the environment. Although various contributions can be found in the literature about sediment yield modeling and bank erosion monitoring, the link between weather conditions, river flow rate and bank erosion remains scarcely known. Thus, a basin scale assessment of sediment yield due to riverbank erosion is an objective hard to be reached. In order to enhance the current knowledge in this field, a monitoring method based on high resolution 3D model reconstruction of riverbanks, surveyed by multi-temporal terrestrial laser scanning, was applied to four banks in Val Tartano, Northern Italy. Six data acquisitions over one year were taken, with the aim to better understand the erosion processes and their triggering factors by means of more frequent observations compared to usual annual campaigns. The objective of the research is to address three key questions concerning bank erosion: “how” erosion happens, “when” during the year and “how much” sediment is eroded. The method proved to be effective and able to measure both eroded and deposited volume in the surveyed area. Finally an attempt to extrapolate basin scale volume for bank erosion is presented.

  4. MEASURING LEAF WATER CONTENT USING MULTISPECTRAL TERRESTRIAL LASER SCANNING

    Directory of Open Access Journals (Sweden)

    S. Junttila

    2017-10-01

    Full Text Available Climate change is increasing the amount and intensity of disturbance events, i.e. drought, pest insect outbreaks and fungal pathogens, in forests worldwide. Leaf water content (LWC is an early indicator of tree stress that can be measured remotely using multispectral terrestrial laser scanning (MS-TLS. LWC affects leaf reflectance in the shortwave infrared spectrum which can be used to predict LWC from spatially explicit MS-TLS intensity data. Here, we investigated the relationship between LWC and MS-TLS intensity features at 690 nm, 905 nm and 1550 nm wavelengths with Norway spruce seedlings in greenhouse conditions. We found that a simple ratio of 905 nm and 1550 nm wavelengths was able to explain 84 % of the variation (R2 in LWC with a respective prediction accuracy of 0.0041 g/cm2. Our results showed that MS-TLS can be used to estimate LWC with a reasonable accuracy in environmentally stable conditions.

  5. Wetland Microtopographic Structure is Revealed with Terrestrial Laser Scanning

    Science.gov (United States)

    Diamond, J.; Stovall, A. E.; Mclaughlin, D. L.; Slesak, R.

    2017-12-01

    Wetland microtopographic structure and its function has been the subject of research for decades, and several investigations suggest that microtopography is generated by autogenic ecohydrologic processes. But due to the difficulty of capturing the true spatial variability of wetland microtopography, many of the hypotheses for self-organization have remained elusive to test. We employ a novel method of Terrestrial Laser Scanning (TLS) that reveals an unprecedented high-resolution (structure of wetland microtopography in 10 black ash (Fraxinus nigra) stands of northern Minnesota, USA. Here we present the first efforts to synthesize this information and show that TLS provides a good representation of real microtopographic structure, where TLS accurately measured hummock height, but occlusion of low points led to a slight negative bias. We further show that TLS can accurately locate microtopographic high points (hummocks), as well as estimate their height and area. Using these new data, we estimate distributions in both microtopographic elevation and hummock area in each wetland and relate these to monitored hydrologic regime; in doing so, we test hypotheses linking emergent microtopographic patterns to putative hydrologic controls. Finally, we discuss future efforts to enumerate consequent influences of microtopography on wetland systems (soil properties and vegetation composition).

  6. Detecting Terrain Stoniness From Airborne Laser Scanning Data †

    Directory of Open Access Journals (Sweden)

    Paavo Nevalainen

    2016-08-01

    Full Text Available Three methods to estimate the presence of ground surface stones from publicly available Airborne Laser Scanning (ALS point clouds are presented. The first method approximates the local curvature by local linear multi-scale fitting, and the second method uses Discrete-Differential Gaussian curvature based on the ground surface triangulation. The third baseline method applies Laplace filtering to Digital Elevation Model (DEM in a 2 m regular grid data. All methods produce an approximate Gaussian curvature distribution which is then vectorized and classified by logistic regression. Two training data sets consisted of 88 and 674 polygons of mass-flow deposits, respectively. The locality of the polygon samples is a sparse canopy boreal forest, where the density of ALS ground returns is sufficiently high to reveal information about terrain micro-topography. The surface stoniness of each polygon sample was categorized for supervised learning by expert observation on the site. The leave-pair-out (L2O cross-validation of the local linear fit method results in the area under curve A U C = 0 . 74 and A U C = 0 . 85 on two data sets, respectively. This performance can be expected to suit real world applications such as detecting coarse-grained sediments for infrastructure construction. A wall-to-wall predictor based on the study was demonstrated.

  7. Laser scanning measurements on trees for logging harvesting operations.

    Science.gov (United States)

    Zheng, Yili; Liu, Jinhao; Wang, Dian; Yang, Ruixi

    2012-01-01

    Logging harvesters represent a set of high-performance modern forestry machinery, which can finish a series of continuous operations such as felling, delimbing, peeling, bucking and so forth with human intervention. It is found by experiment that during the process of the alignment of the harvesting head to capture the trunk, the operator needs a lot of observation, judgment and repeated operations, which lead to the time and fuel losses. In order to improve the operation efficiency and reduce the operating costs, the point clouds for standing trees are collected with a low-cost 2D laser scanner. A cluster extracting algorithm and filtering algorithm are used to classify each trunk from the point cloud. On the assumption that every cross section of the target trunk is approximate a standard circle and combining the information of an Attitude and Heading Reference System, the radii and center locations of the trunks in the scanning range are calculated by the Fletcher-Reeves conjugate gradient algorithm. The method is validated through experiments in an aspen forest, and the optimized calculation time consumption is compared with the previous work of other researchers. Moreover, the implementation of the calculation result for automotive capturing trunks by the harvesting head during the logging operation is discussed in particular.

  8. Detection of a fluorescent-labeled avidin-nucleic acid nanoassembly by confocal laser endomicroscopy in the microvasculature of chronically inflamed intestinal mucosa

    Directory of Open Access Journals (Sweden)

    Buda A

    2015-01-01

    Full Text Available Andrea Buda,1,* Sonia Facchin,1,* Elisa Dassie,2 Elisabetta Casarin,3 Mark A Jepson,4 Helmut Neumann,5 Giorgia Hatem,1 Stefano Realdon,6 Renata D’Incà,1 Giacomo Carlo Sturniolo,1 Margherita Morpurgo3 1Department of Surgical, Oncological, and Gastroenterological Sciences, University of Padova, 2Department of Molecular Medicine, University of Padova, Padova, Italy; 3Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, Italy; 4School of Biochemistry and Wolfson Bioimaging Facility, University of Bristol, Bristol, UK; 5Ludwig Demlig Endoscopic Center of Excellence, ESGE Endoscopy Training Center, University of Erlangen-Nuremberg, Erlangen, Germany; 6Veneto Institute of Oncology IOV-IRCCS, Padova, Italy *These authors contributed equally to this work Abstract: Inflammatory bowel diseases are chronic gastrointestinal pathologies causing great discomfort in both children and adults. The pathogenesis of inflammatory bowel diseases is not yet fully understood and their diagnosis and treatment are often challenging. Nanoparticle-based strategies have been tested in local drug delivery to the inflamed colon. Here, we have investigated the use of the novel avidin-nucleic acid nanoassembly (ANANAS platform as a potential diagnostic carrier in an experimental model of inflammatory bowel diseases. Fluorescent-labeled ANANAS nanoparticles were administered to mice with chemically induced chronic inflammation of the large intestine. Localization of mucosal nanoparticles was assessed in vivo by dual-band confocal laser endomicroscopy. This technique enables characterization of the mucosal microvasculature and crypt architecture at subcellular resolution. Intravascular nanoparticle distribution was observed in the inflamed mucosa but not in healthy controls, demonstrating the utility of the combination of ANANAS and confocal laser endomicroscopy for highlighting intestinal inflammatory conditions. The specific localization of

  9. Confocal microscopy studies of morphology and apoptosis: ovaries, limbs, embryos and insects

    Science.gov (United States)

    Confocal laser scanning microscopy (CLSM) is a technique that is capable of generating serial sections of whole-mount tissue and then reassembling the computer-stored images as a virtual 3-dimensional structure. In many ways CLSM offers an alternative to traditional sectioning ap...

  10. Confocal microscopy of thick tissue sections: 3D visualizaiton of rat kidney glomeruli

    Science.gov (United States)

    Confocal laser scanning microscopy (CLSM) as a technique capable of generating serial sections of whole-mount tissue and then reassembling the computer-acquired images as a virtual 3-dimentional structure. In many ways CLSM offers an alternative to traditional sectioning approac...

  11. Confocal Microscopy of thick tissue sections: 3D Visualization of rat kidney glomeruli

    Science.gov (United States)

    Confocal laser scanning microscopy (CLSM) as a technique capable of generating serial sections of whole-mount tissue and then reassembling the computer-acquired images as a virtual 3-dimentional structure. In many ways CLSM offers an alternative to traditional sectioning approac...

  12. Insight into the Microbial Multicellular Lifestyle via Flow-Cell Technology and Confocal Microscopy

    DEFF Research Database (Denmark)

    Pamp, Sünje Johanna; Sternberg, Claus; Tolker-Nielsen, Tim

    2009-01-01

    , industry, and human health. Accordingly a number of biofilm model systems, molecular tools, microscopic techniques, and image analysis programs have been employed for the study of biofilms under controlled and reproducible conditions. Studies using confocal laser scanning microscopy (CLSM) of biofilms...

  13. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: FOUNDATIONS FOR QUANTIFYING CYTOMETRIC APPLICATIONS WITH SPECTROSCOPIC INSTRUMENTS

    Science.gov (United States)

    The confocal laser-scanning microscopy (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 the emitted signal. The accuracy of these measurements demands t...

  14. An FFT-based Method for Attenuation Correction in Fluorescence Confocal Microscopy

    NARCIS (Netherlands)

    Roerdink, J.B.T.M.; Bakker, M.

    1993-01-01

    A problem in three-dimensional imaging by a confocal scanning laser microscope (CSLM) in the (epi)fluorescence mode is the darkening of the deeper layers due to absorption and scattering of both the excitation and the fluorescence light. In this paper we propose a new method to correct for these

  15. RANSAC approach for automated registration of terrestrial laser scans using linear features

    Directory of Open Access Journals (Sweden)

    K. Al-Durgham

    2013-10-01

    Full Text Available The registration process of terrestrial laser scans (TLS targets the problem of how to combine several laser scans in order to attain better information about features than what could be obtained through single scan. The main goal of the registration process is to estimate the parameters which determine geometrical variation between the origins of datasets collected from different locations. Scale, shifts, and rotation parameters are usually used to describe such variation. This paper presents a framework for the registration of overlapping terrestrial laser scans by establishing an automatic matching strategy that uses 3D linear features. More specifically, invariant separation characteristics between 3D linear features extracted from laser scans will be used to establish hypothesized conjugate linear features between the laser scans. These candidate matches are then used to geo-reference scans relative to a common reference frame. The registration workflow simulates the well-known RANndom Sample Consensus method (RANSAC for determining the registration parameters, whereas the iterative closest projected point (ICPP is utilized to determine the most probable solution of the transformation parameters from several solutions. The experimental results prove that the proposed methodology can be used for the automatic registration of terrestrial laser scans using linear features.

  16. Development of Smart Precision Forest in Conifer Plantation in Japan Using Laser Scanning Data

    Science.gov (United States)

    Katoh, M.; Deng, S.; Takenaka, Y.; Cheung, K.; Oono, K.; Horisawa, M.; Hyyppä, J.; Yu, X.; Liang, X.; Wang, Y.

    2017-10-01

    Currently, the authors are planning to launch a consortium effort toward Japan's first smart precision forestry project using laser data and to develop this technology throughout the country. Smart precision forestry information gathered using the Nagano model (laser scanning from aircraft, drone, and backpack) is being developed to improve the sophistication of forest information, reduce labor-intensive work, maintain sustainable timber productivity, and facilitate supply chain management by laser sensing information in collaboration with industry, academia, and government. In this paper, we outline the research project and the technical development situation of unmanned aerial vehicle laser scanning.

  17. DEVELOPMENT OF SMART PRECISION FOREST IN CONIFER PLANTATION IN JAPAN USING LASER SCANNING DATA

    Directory of Open Access Journals (Sweden)

    M. Katoh

    2017-10-01

    Full Text Available Currently, the authors are planning to launch a consortium effort toward Japan’s first smart precision forestry project using laser data and to develop this technology throughout the country. Smart precision forestry information gathered using the Nagano model (laser scanning from aircraft, drone, and backpack is being developed to improve the sophistication of forest information, reduce labor-intensive work, maintain sustainable timber productivity, and facilitate supply chain management by laser sensing information in collaboration with industry, academia, and government. In this paper, we outline the research project and the technical development situation of unmanned aerial vehicle laser scanning.

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

    Science.gov (United States)

    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.

  19. Periodic and uniform nanogratings formed on cemented carbide by femtosecond laser scanning

    International Nuclear Information System (INIS)

    Lian, Yunsong; Deng, Jianxin; Xing, Youqiang; Lei, Shuting; Yu, Xiaoming

    2013-01-01

    Periodic and uniform nanogratings are fabricated by femtosecond laser scanning on cemented carbide. Specifically, three experiments are designed to study the influence of single pulse energy, scanning speed, and scanning spacing on the period and the uniformity of the formed nanogratings. The results show that the sample with single pulse energy of 2 μJ, scanning speed of 1000 μm/s, and scanning spacing of 5 μm shows the best quality of nanogratings among all the tested samples at different processing parameters. The uniformity of the nanogratings is largely determined by single pulse energy, scanning speed, and scanning spacing. Single pulse energy and scanning speed significantly affect the period of the nanogratings, whereas the period of the nanogratings maintains a fixed value under different scanning spacings. The period of the nanogratings increases gradually with the decrease of the single pulse energy and the increase of the scanning speed, respectively.

  20. Retinal Oximetry with Scanning Laser Ophthalmoscope in Infants.

    Directory of Open Access Journals (Sweden)

    Wouter B Vehmeijer

    Full Text Available Dual wavelength retinal oximetry has been developed for adults, but is not available for infants. Retinal oximetry may provide insight into the pathophysiology of oxygen-mediated diseases like retinopathy of prematurity. More insight in the oxygen metabolism of the retina in infants may provide valuable clues for better understanding and subsequent prevention or treatment of the disease. The measurements of oxygen saturation are obtained with two fundus images simultaneously captured in two different wavelengths of light. The comparison in light absorption of oxygenated and deoxygenated hemoglobin can be used to estimate the oxygen saturation within the retinal vessels by means of a software algorithm. This study aims to make retinal oximetry available for neonates. The first step towards estimating retinal oxygen saturation is determining the optical density ratio. Therefore, the purpose of this study is to image healthy newborn infants with a scanning laser ophthalmoscope and determine the optical density ratio for retinal oximetry analysis.Images of the retina of full-term healthy infants were obtained with an SLO, Optomap 200Tx (Optos, with two laser wavelengths (532nm and 633nm. The infant lay face down on the lower arm of the parent, while the parent supported the chest and chin with one hand, and stabilized the back with the other hand. No mydriatics or eyelid specula were used during this study. The images were analyzed with modified Oxymap Analyzer software for calculation of the Optical Density Ratio (ODR and vessel width. The ODR is inversely and approximately linearly related to the oxygen saturation. Measurements were included from the superotemporal vessel pair. A paired t-test was used for statistical analysis.Fifty-nine infants, (58% female, were included with mean gestational age of 40 ± 1.3 weeks (mean ± SD and mean post-natal age of 16 ± 4.8 days. A total of 28 images were selected for retinal oximetry analysis. The ODR was

  1. Quantification of aggregate grain shape characteristics using 3-D laser scanning technology

    CSIR Research Space (South Africa)

    Mgangira, Martin B

    2013-07-01

    Full Text Available to identify the differences between individual aggregates. It was possible to quantify differences in particle shape characteristics at the small particle scale. The study has demonstrated the advantages of the innovative 3-D laser scanning technology...

  2. Three-dimensional laser scanning technique to quantify aggregate and ballast shape properties

    CSIR Research Space (South Africa)

    Anochie-Boateng, Joseph

    2013-06-01

    Full Text Available methods towards a more accurate and automated techniques to quantify aggregate shape properties. This paper validates a new flakiness index equation using three-dimensional (3-D) laser scanning data of aggregate and ballast materials obtained from...

  3. [Results of therapy of children with amblyopia by scanning stimulating laser].

    Science.gov (United States)

    Chentsova, O B; Magaramova, M D; Grechanyĭ, M P

    1997-01-01

    A new effective method for the treatment of amblyopia was used in 113 children: stimulation with ophthalmological SLSO-208A scanning laser by two methods differing by the transmission coefficient and scanning pattern. Good results were attained, the best when laser exposure was combined with traditional therapy for amblyopia and in the patients with the central fixation. The results were assessed by the main parameters of visual functions and the stability of the effect.

  4. Demonstration of intermediate cells during human prostate epithelial differentiation in situ and in vitro using triple-staining confocal scanning microscopy.

    Science.gov (United States)

    van Leenders, G; Dijkman, H; Hulsbergen-van de Kaa, C; Ruiter, D; Schalken, J

    2000-08-01

    In human prostate epithelium, morphologically basal and luminal cells can be discriminated. The basal cell layer that putatively contains progenitor cells of the secretory epithelium is characterized by the expression of keratins (K) 5 and 14. Luminal cells represent the secretory compartment of the epithelium and express K8 and 18. We developed a technique for the simultaneous analysis of K5, 14, and 18 to identify intermediate cell stages in the prostate epithelium and to study the dynamic aspects of its differentiation in vitro. Nonmalignant prostate tissue and primary epithelial cultures were immunohistochemically characterized using triple staining with antibodies for K5, K14, and K18. Antibodies for K18 and K5 were conjugated directly with fluorochromes Alexa 488 and 546. K14 was visualized indirectly with streptavidin-Cy5. Keratin expression was analyzed by confocal scanning microscopy. The occurrence of exocrine and neuroendocrine differentiation in culture was determined via antibodies to prostate-specific antigen (PSA), chromogranin A, and serotonin. We found that basal cells expressed either K5(++)/14(++)/18+ or K5(++)/18+. The majority of luminal cells expressed K18(++), but colocalization of K5+/18(++) were recognized. Epithelial monolayer cultures predominantly revealed the basal cell phenotype K5(++)/14(++)/18+, whereas intermediate subpopulations expressing K5+/14+/18(++) and K5+/18(++) were also identified. On confluence, differentiation was induced as multicellular gland-like buds, and extensions became evident on top of the monolayer. These structures were composed of K18(++)- and K5+/18(+)-positive cell clusters surrounded by phenotypically basal cells. Few multicellular structures and cells in the monolayer showed exocrine differentiation (PSA+), but expression of chromogranin A and serotonin was absent. We conclude that simultaneous evaluation of keratin expression is useful for analyzing epithelial differentiation in the prostate. During this

  5. Application of Mobile Laser Scanning for Lean and Rapid Highway Maintenance and Construction

    Science.gov (United States)

    2015-08-28

    Mobile Terrestrial Laser Scanning (MTLS) is an emerging technology that combines the use of a laser scanner(s), the Global Navigation Satellite System (GNSS), and an Inertial Measurement Unit (IMU) on a vehicle to collect geo-spatial data. The overal...

  6. An improved three-dimensional non-scanning laser imaging system based on digital micromirror device

    Science.gov (United States)

    Xia, Wenze; Han, Shaokun; Lei, Jieyu; Zhai, Yu; Timofeev, Alexander N.

    2018-01-01

    Nowadays, there are two main methods to realize three-dimensional non-scanning laser imaging detection, which are detection method based on APD and detection method based on Streak Tube. However, the detection method based on APD possesses some disadvantages, such as small number of pixels, big pixel interval and complex supporting circuit. The detection method based on Streak Tube possesses some disadvantages, such as big volume, bad reliability and high cost. In order to resolve the above questions, this paper proposes an improved three-dimensional non-scanning laser imaging system based on Digital Micromirror Device. In this imaging system, accurate control of laser beams and compact design of imaging structure are realized by several quarter-wave plates and a polarizing beam splitter. The remapping fiber optics is used to sample the image plane of receiving optical lens, and transform the image into line light resource, which can realize the non-scanning imaging principle. The Digital Micromirror Device is used to convert laser pulses from temporal domain to spatial domain. The CCD with strong sensitivity is used to detect the final reflected laser pulses. In this paper, we also use an algorithm which is used to simulate this improved laser imaging system. In the last, the simulated imaging experiment demonstrates that this improved laser imaging system can realize three-dimensional non-scanning laser imaging detection.

  7. Assessment of Relative Accuracy of AHN-2 Laser Scanning Data Using Planar Features

    NARCIS (Netherlands)

    Khoshelham, K.; Soudarissanane, S.; Van der Sande, C.

    2010-01-01

    AHN-2 is the second part of the Actueel Hoogtebestand Nederland project, which concerns the acquisition of high-resolution altimetry data over the entire Netherlands using airborne laser scanning. The accuracy assessment of laser altimetry data usually relies on comparing corresponding tie elements,

  8. Real-time high dynamic range laser scanning microscopy

    Science.gov (United States)

    Vinegoni, C.; Leon Swisher, C.; Fumene Feruglio, P.; Giedt, R. J.; Rousso, D. L.; Stapleton, S.; Weissleder, R.

    2016-04-01

    In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging.

  9. Laser line scan underwater imaging by complementary metal-oxide-semiconductor camera

    Science.gov (United States)

    He, Zhiyi; Luo, Meixing; Song, Xiyu; Wang, Dundong; He, Ning

    2017-12-01

    This work employs the complementary metal-oxide-semiconductor (CMOS) camera to acquire images in a scanning manner for laser line scan (LLS) underwater imaging to alleviate backscatter impact of seawater. Two operating features of the CMOS camera, namely the region of interest (ROI) and rolling shutter, can be utilized to perform image scan without the difficulty of translating the receiver above the target as the traditional LLS imaging systems have. By the dynamically reconfigurable ROI of an industrial CMOS camera, we evenly divided the image into five subareas along the pixel rows and then scanned them by changing the ROI region automatically under the synchronous illumination by the fun beams of the lasers. Another scanning method was explored by the rolling shutter operation of the CMOS camera. The fun beam lasers were turned on/off to illuminate the narrow zones on the target in a good correspondence to the exposure lines during the rolling procedure of the camera's electronic shutter. The frame synchronization between the image scan and the laser beam sweep may be achieved by either the strobe lighting output pulse or the external triggering pulse of the industrial camera. Comparison between the scanning and nonscanning images shows that contrast of the underwater image can be improved by our LLS imaging techniques, with higher stability and feasibility than the mechanically controlled scanning method.

  10. Europium Uptake and Partitioning in Oat (Avena sativa) Roots as studied By Laser-Induced Fluorescence Spectroscopy and Confocal Microscopy Profiling Technique

    International Nuclear Information System (INIS)

    Fellows, Robert J.; Wang, Zheming; Ainsworth, Calvin C.

    2003-01-01

    The uptake of Eu3+ by elongating oat plant roots was studied by fluorescence spectroscopy, fluorescence lifetime measurement, as well as laser excitation time-resolved confocal fluorescence profiling technique. The results of this work indicated that the initial uptake of Eu(III) by oat root was most evident within the apical meristem of the root just proximal to the root cap. Distribution of assimilated Eu(III) within the roots differentiation and elongation zone was non-uniform. Higher concentrations were observed within the vascular cylinder, specifically in the phloem and developing xylem parenchyma. Elevated levels of the metal were also observed in the root hairs of the mature root. The concentration of assimilated Eu3+ dropped sharply from the apical meristem to the differentiation and elongation zone and then gradually decreased as the distance from the root cap increased. Fluorescence spectroscopic characteristics of the assimilated Eu3+ suggested that the Eu3+ exists a s inner-sphere mononuclear complexes inside the root. This work has also demonstrated the effectiveness of a time-resolved Eu3+ fluorescence spectroscopy and confocal fluorescence profiling techniques for the in vivo, real-time study of metal[Eu3+] accumulation by a functioning intact plant root. This approach can prove valuable for basic and applied studies in plant nutrition and environmental uptake of actinide radionuclides

  11. Distance measurement using frequency scanning interferometry with mode-hoped laser

    Science.gov (United States)

    Medhat, M.; Sobee, M.; Hussein, H. M.; Terra, O.

    2016-06-01

    In this paper, frequency scanning interferometry is implemented to measure distances up to 5 m absolutely. The setup consists of a Michelson interferometer, an external cavity tunable diode laser, and an ultra-low expansion (ULE) Fabry-Pérot (FP) cavity to measure the frequency scanning range. The distance is measured by acquiring simultaneously the interference fringes from, the Michelson and the FP interferometers, while scanning the laser frequency. An online fringe processing technique is developed to calculate the distance from the fringe ratio while removing the parts result from the laser mode-hops without significantly affecting the measurement accuracy. This fringe processing method enables accurate distance measurements up to 5 m with measurements repeatability ±3.9×10-6 L. An accurate translation stage is used to find the FP cavity free-spectral-range and therefore allow accurate measurement. Finally, the setup is applied for the short distance calibration of a laser distance meter (LDM).

  12. 3D laser scanning in civil engineering - measurements of volume of earth masses

    Science.gov (United States)

    Pawłowicz, J. A.; Szafranko, E.; Harasymiuk, J.

    2018-03-01

    Considering the constant drive to improve and accelerate building processes as well as possible applications of the latest technological achievements in civil engineering practice, the author has proposed to use 3D laser scanning in the construction industry. For example, data achieved through a 3D laser scanning process will facilitate making inventories of parameters of buildings in a very short time, will enable one to check irregularly shaped masses of earth, heavy and practically impossible to calculate precisely using traditional techniques. The other part of the research, performed in the laboratory, consisted of measurements of a model mound of earth. All the measurements were made with a 3D SkanStation C10 laser scanner manufactured by Leica. The data were analyzed. The results suggest that there are great opportunities for using the laser scanning technology in civil engineering

  13. 3D Laser Scanning Assisted by Ordinary Plane Mirror for Non-direct Viewing Area

    Directory of Open Access Journals (Sweden)

    ZHANG Fan

    2017-12-01

    Full Text Available Terrestrial 3D laser scanning is one of principal methods to get the geometric information of object surface,and the integrity of the scanned object is a basic requirement in data acquisition. In order to solve the missing point cloud problem due to the scanning dead angle caused by confined working space,this paper proposes a method using ordinary plane mirror to obtain laser scanning data for non-direct viewing area according to the plane mirror reflection principle,analyzes the influence mechanism of the ordinary plane mirror on the propagation path and distance of laser beam,deduces the coordinate equation of the object point corresponding to the image point reflected by ordinary plane mirror in laser scanning. Given the laser scanning characteristic,this paper introduces a mirror reflection system included target balls and ordinary plane mirror,and expounds the system construction,system calibration and constructing method of system coordinate system. The feasibility and precision of the method are verified by experiments.

  14. Distribution and avoidance of debris on epoxy resin during UV ns-laser scanning processes

    Science.gov (United States)

    Veltrup, Markus; Lukasczyk, Thomas; Ihde, Jörg; Mayer, Bernd

    2018-05-01

    In this paper the distribution of debris generated by a nanosecond UV laser (248 nm) on epoxy resin and the prevention of the corresponding re-deposition effects by parameter selection for a ns-laser scanning process were investigated. In order to understand the mechanisms behind the debris generation, in-situ particle measurements were performed during laser treatment. These measurements enabled the determination of the ablation threshold of the epoxy resin as well as the particle density and size distribution in relation to the applied laser parameters. The experiments showed that it is possible to reduce debris on the surface with an adapted selection of pulse overlap with respect to laser fluence. A theoretical model for the parameter selection was developed and tested. Based on this model, the correct choice of laser parameters with reduced laser fluence resulted in a surface without any re-deposited micro-particles.

  15. Confocal Microscopy

    Science.gov (United States)

    Liu, Jian; Tan, Jiubin

    2016-12-01

    The confocal microscope is appropriate for imaging cells or the measurement of industrial artefacts. However, junior researchers and instrument users sometimes misuse imaging concepts and metrological characteristics, such as position resolution in industrial metrology and scale resolution in bio-imaging. And, metrological characteristics or influence factors in 3D measurement such as height assessment error caused by 3D coupling effect are so far not yet identified. In this book, the authors outline their practices by the working experiences on standardization and system design. This book assumes little previous knowledge of optics, but rich experience in engineering of industrial measurements, in particular with profile metrology or areal surface topography will be very helpful to understand the theoretical concerns and value of the technological advances. It should be useful for graduate students or researchers as extended reading material, as well as microscope users alongside their handbook.

  16. A vision-based system for fast and accurate laser scanning in robot-assisted phonomicrosurgery.

    Science.gov (United States)

    Dagnino, Giulio; Mattos, Leonardo S; Caldwell, Darwin G

    2015-02-01

    Surgical quality in phonomicrosurgery can be improved by open-loop laser control (e.g., high-speed scanning capabilities) with a robust and accurate closed-loop visual servoing systems. A new vision-based system for laser scanning control during robot-assisted phonomicrosurgery was developed and tested. Laser scanning was accomplished with a dual control strategy, which adds a vision-based trajectory correction phase to a fast open-loop laser controller. The system is designed to eliminate open-loop aiming errors caused by system calibration limitations and by the unpredictable topology of real targets. Evaluation of the new system was performed using CO(2) laser cutting trials on artificial targets and ex-vivo tissue. This system produced accuracy values corresponding to pixel resolution even when smoke created by the laser-target interaction clutters the camera view. In realistic test scenarios, trajectory following RMS errors were reduced by almost 80 % with respect to open-loop system performances, reaching mean error values around 30 μ m and maximum observed errors in the order of 60 μ m. A new vision-based laser microsurgical control system was shown to be effective and promising with significant positive potential impact on the safety and quality of laser microsurgeries.

  17. Histopathological confirmation of similar intramucosal distribution of fluorescein in both intravenous administration and local mucosal application for probe-based confocal laser endomicroscopy of the normal stomach.

    Science.gov (United States)

    Nonaka, Kouichi; Ohata, Ken; Ban, Shinichi; Ichihara, Shin; Takasugi, Rumi; Minato, Yohei; Tashima, Tomoaki; Matsuyama, Yasushi; Takita, Maiko; Matsuhashi, Nobuyuki; Neumann, Helmut

    2015-12-16

    Probe-based confocal laser endomicroscopy (pCLE) is capable of acquiring in vivo magnified cross-section images of the gastric mucosa. Intravenous injection of fluorescein sodium is used for confocal imaging. However, it is still under debate if local administration of the dye to the mucosa is also effective for confocal imaging as it is not yet clear if topical application also reveals the intramucosal distribution of fluorescein. The objective of this study was to evaluate the intramucosal distribution of fluorescein sodium after topical application and to compare the distribution to the conventional intravenous injection used for confocal imaging. pCLE of the stomach uninfected with Helicobacter pylori was performed in a healthy male employing intravenous administration and local mucosal application of fluorescein. The mucosa of the lower gastric body was biopsied 1 min and 5 min after intravenous administration or local mucosal application of fluorescein, and the distribution of fluorescein in the biopsy samples was examined histologically. Green fluorescence was already observed in the cytoplasm of fundic glandular cells in the biopsied deep mucosa 1 min after local mucosal application of fluorescein. It was also observed in the foveolar lumen and inter-foveolar lamina propria, although it was noted at only a few sites. In the tissue biopsied 5 min after the local mucosal application of fluorescein, green fluorescence was more frequently noted in the cytoplasm of fundic glandular cells than in that 1 min after the local mucosal application of fluorescein, although obvious green fluorescence was not identified in the foveolar lumen or inter-foveolar lamina propria. The distribution of intravenously administered fluorescein in the cytoplasm of fundic glandular cells was also clearly observed similarly to that after local mucosal application of fluorescein. Green fluorescence in more cells was observed in many cells 5 min after intravenous administration compared

  18. Structural modification of silica glass by laser scanning

    International Nuclear Information System (INIS)

    Zhao Jian; Sullivan, James; Zayac, John; Bennett, Ted D.

    2004-01-01

    The thermophysical nature of rapid CO 2 laser heating of silica glass is explored using a numerical simulation that considers the structural state of the glass, as characterized by the fictive temperature. The fictive temperature reflects the thermodynamic temperature at which the glass structure would be in equilibrium. To demonstrate that the thermophysical model can accurately predict the structural change in the glass, the fictive temperature is measured experimentally utilizing the fact that the fictive temperature change corresponds to a change of glass properties that can be revealed through wet chemical etching. The relationship between the etch rate and the fictive temperature is determined by preparing and etching samples of known fictive temperature. Wet chemical etching is used to measure the fictive temperature over the entire laser affected zone and the results are found to compare favorably with the results of the thermophysical model. The model and experimental measurements demonstrate that rapid laser processing results in an increased fictive temperature near the surface of the glass. The fictive temperature increase is about 1000 K and is uniform to within 5% over the laser affected zone. Near the boundary of this zone, the fictive temperature transitions abruptly to the value of the surrounding untreated glass

  19. Unsynchronized scanning with a low-cost laser range finder for real-time range imaging

    Science.gov (United States)

    Hatipoglu, Isa; Nakhmani, Arie

    2017-06-01

    Range imaging plays an essential role in many fields: 3D modeling, robotics, heritage, agriculture, forestry, reverse engineering. One of the most popular range-measuring technologies is laser scanner due to its several advantages: long range, high precision, real-time measurement capabilities, and no dependence on lighting conditions. However, laser scanners are very costly. Their high cost prevents widespread use in applications. Due to the latest developments in technology, now, low-cost, reliable, faster, and light-weight 1D laser range finders (LRFs) are available. A low-cost 1D LRF with a scanning mechanism, providing the ability of laser beam steering for additional dimensions, enables to capture a depth map. In this work, we present an unsynchronized scanning with a low-cost LRF to decrease scanning period and reduce vibrations caused by stop-scan in synchronized scanning. Moreover, we developed an algorithm for alignment of unsynchronized raw data and proposed range image post-processing framework. The proposed technique enables to have a range imaging system for a fraction of the price of its counterparts. The results prove that the proposed method can fulfill the need for a low-cost laser scanning for range imaging for static environments because the most significant limitation of the method is the scanning period which is about 2 minutes for 55,000 range points (resolution of 250x220 image). In contrast, scanning the same image takes around 4 minutes in synchronized scanning. Once faster, longer range, and narrow beam LRFs are available, the methods proposed in this work can produce better results.

  20. The geometry of terrestrial laser scanning; identification of errors, modeling and mitigation of scanning geometry

    NARCIS (Netherlands)

    Soudarissanane, S.S.

    2016-01-01

    Over the past few decades, Terrestrial Laser Scanners are increasingly being used in a broad spectrum of applications, from surveying to civil engineering, medical modeling and forensics. Especially surveying applications require on one hand a quickly obtainable, high resolution point cloud but also

  1. High-efficient Nd:YAG microchip laser for optical surface scanning

    Science.gov (United States)

    Šulc, Jan; Jelínková, Helena; Nejezchleb, Karel; Škoda, Václav

    2017-12-01

    A CW operating, compact, high-power, high-efficient diode pumped 1064nm laser, based on Nd:YAG active medium, was developed for optical surface scanning and mapping applications. To enhance the output beam quality, laser stability, and compactness, a microchip configuration was used. In this arrangement the resonator mirrors were deposited directly on to the laser crystal faces. The Nd-doping concentration was 1 at.% Nd/Y. The Nd:YAG crystal was 5mm long. The laser resonator without pumping radiation recuperation was investigated {the output coupler was transparent for pumping radiation. For the generated laser radiation the output coupler reflectivity was 95%@1064 nm. The diameter of the samples was 5 mm. For the laser pumping two arrangements were investigated. Firstly, a fibre coupled laser diode operating at wavelength 808nm was used in CW mode. The 400 ¹m fiber was delivering up to 14W of pump power amplitude to the microchip laser. The maximum CW output power of 7.2W @ 1064nm in close to TEM00 beam was obtained for incident pumping power 13.7W @ 808 nm. The differential efficiency in respect to the incident pump power reached 56 %. Secondly, a single-emitter, 1W laser diode operating at 808nm was used for Nd:YAG microchip pumping. The laser pumping was directly coupled into the microchip laser using free-space lens optics. Slope efficiency up to 70% was obtained in stable, high-quality, 1064nm laser beam with CW power up to 350mW. The system was successfully used for scanning of super-Gaussian laser mirrors reflectivity profile.

  2. Three-Dimensional Laser Scanning for Geometry Documentation and Construction Management of Highway Tunnels during Excavation

    Science.gov (United States)

    Gikas, Vassilis

    2012-01-01

    Driven by progress in sensor technology, computer software and data processing capabilities, terrestrial laser scanning has recently proved a revolutionary technique for high accuracy, 3D mapping and documentation of physical scenarios and man-made structures. Particularly, this is of great importance in the underground space and tunnel construction environment as surveying engineering operations have a great impact on both technical and economic aspects of a project. This paper discusses the use and explores the potential of laser scanning technology to accurately track excavation and construction activities of highway tunnels. It provides a detailed overview of the static laser scanning method, its principles of operation and applications for tunnel construction operations. Also, it discusses the planning, execution, data processing and analysis phases of laser scanning activities, with emphasis given on geo-referencing, mesh model generation and cross-section extraction. Specific case studies are considered based on two construction sites in Greece. Particularly, the potential of the method is examined for checking the tunnel profile, producing volume computations and validating the smoothness/thickness of shotcrete layers at an excavation stage and during the completion of excavation support and primary lining. An additional example of the use of the method in the geometric documentation of the concrete lining formwork is examined and comparisons against dimensional tolerances are examined. Experimental comparisons and analyses of the laser scanning method against conventional surveying techniques are also considered. PMID:23112655

  3. SOME ASPECTS OF SCANNING LASER OPHTHALMOSCOPY IN THE DIAGNOSTICS OF OPHTHALMOPATHOLOGY

    Directory of Open Access Journals (Sweden)

    S. A. Kochergin

    2017-01-01

    Full Text Available The exact diagnosis of the fundus pathology requires the most modern equipment use. This is mandatory for the selection of the most complete therapy and monitoring of ongoing treatment. At present, the method of scanning laser ophthalmoscopy is widely spread. However, for the earliest detection of the smallest pathological changes, data of the normal ocular fundus state using a scanning laser ophthalmoscope is necessary. Thus, the purpose of our research becomes relevant. Purpose: to give a characteristic of the fundus in patients without concomitant pathology with using various modes of a scanning laser ophthalmoscope. Patients and methods. 116 people (232 eyes at the age from 17 to 71 years (mean age 32.5±12 years were examined. The patients were divided into two groups. Group I: 81 patients (162 eyes with different ophthalmopathology. Group II: 35 people (70 eyes — practically healthy and did not have an anamnesis of consulting an ophthalmologist. Diagnosis of the patients’ fundus was performed using a scanning laser ophthalmoscopy with retro-mode imaging and autofluorescence registration. Results. After the conducted research features and regularities of the reflectivity distribution of laser beams from the fundus structures are revealed. Also a characteristic of various anatomical formations and zones of the fundus in the normal conditions is given when examined by a scanning laser ophthalmoscope. An algorithm for examining patients and analyzing the images was developed. Conclusion. The use of scanning laser ophthalmoscopy made possible to take a fresh look at the algorithms of diagnosing patients with fundus pathology. Understanding the normal conditions ofundus allowed an earlier detection of the smallest pathological changes in the retina. 

  4. Experimental validation of a newly designed 6 degrees of freedom scanning laser head: Application to three-dimensional beam structure

    International Nuclear Information System (INIS)

    Di Maio, D.; Copertaro, E.

    2013-01-01

    A new scanning laser head is designed to use single Laser Doppler Vibrometer (LDV) for performing measurements up to 6 degrees of freedom (DOF) at a target. The scanning head is supported by a rotating hollow shaft, which allows the laser beam to travel up to the scanning head from an opposite direction where an LDV is set up. The scanning head is made of a set of two mirrors, which deflects the laser beam with an angle so that the rotation of the scanning head produces a conical scan. When measurements are performed at the focal point of the conical scan then three translational vibration components can be measured, otherwise the very small circle scan, before and after the focal point, can measure up to 6 degrees of freedom, including three translations and three rotations. This paper presents the 6DOF scanning head and the measurements of 3D operational deflection shapes of a test structure

  5. Slope excavation quality assessment and excavated volume calculation in hydraulic projects based on laser scanning technology

    Directory of Open Access Journals (Sweden)

    Chao Hu

    2015-04-01

    Full Text Available Slope excavation is one of the most crucial steps in the construction of a hydraulic project. Excavation project quality assessment and excavated volume calculation are critical in construction management. The positioning of excavation projects using traditional instruments is inefficient and may cause error. To improve the efficiency and precision of calculation and assessment, three-dimensional laser scanning technology was used for slope excavation quality assessment. An efficient data acquisition, processing, and management workflow was presented in this study. Based on the quality control indices, including the average gradient, slope toe elevation, and overbreak and underbreak, cross-sectional quality assessment and holistic quality assessment methods were proposed to assess the slope excavation quality with laser-scanned data. An algorithm was also presented to calculate the excavated volume with laser-scanned data. A field application and a laboratory experiment were carried out to verify the feasibility of these methods for excavation quality assessment and excavated volume calculation. The results show that the quality assessment indices can be obtained rapidly and accurately with design parameters and scanned data, and the results of holistic quality assessment are consistent with those of cross-sectional quality assessment. In addition, the time consumption in excavation quality assessment with the laser scanning technology can be reduced by 70%–90%, as compared with the traditional method. The excavated volume calculated with the scanned data only slightly differs from measured data, demonstrating the applicability of the excavated volume calculation method presented in this study.

  6. Towards Robust Self-Calibration for Handheld 3d Line Laser Scanning

    Science.gov (United States)

    Bleier, M.; Nüchter, A.

    2017-11-01

    This paper studies self-calibration of a structured light system, which reconstructs 3D information using video from a static consumer camera and a handheld cross line laser projector. Intersections between the individual laser curves and geometric constraints on the relative position of the laser planes are exploited to achieve dense 3D reconstruction. This is possible without any prior knowledge of the movement of the projector. However, inaccurrately extracted laser lines introduce noise in the detected intersection positions and therefore distort the reconstruction result. Furthermore, when scanning objects with specular reflections, such as glossy painted or metalic surfaces, the reflections are often extracted from the camera image as erroneous laser curves. In this paper we investiagte how robust estimates of the parameters of the laser planes can be obtained despite of noisy detections.

  7. Purinergic receptors have different effects in rat exocrine pancreas. Calcium signals monitored by fura-2 using confocal microscopy

    DEFF Research Database (Denmark)

    Novak, Ivana; Nitschke, Roland; Amstrup, Jan

    2002-01-01

    Pancreatic ducts have several types of purinergic P2 receptors, however, nothing is known about P2 receptors in acini. The aim was to establish whether acini express functional P2 receptors coupled to intracellular Ca2+ signals and to measure the signals ratiometrically in a confocal laser scanning...

  8. Cellular scanning strategy for selective laser melting: Generating reliable, optimized scanning paths and processing parameters

    DEFF Research Database (Denmark)

    Mohanty, Sankhya; Hattel, Jesper Henri

    2015-01-01

    method based uncertainty and reliability analysis. The reliability of the scanning paths are established using cumulative probability distribution functions for process output criteria such as sample density, thermal homogeneity, etc. A customized genetic algorithm is used along with the simulation model...

  9. Performance verification of focus variation and confocal microscopes measuring tilted ultra-fine surfaces

    DEFF Research Database (Denmark)

    Quagliotti, Danilo; Baruffi, Federico; Tosello, Guido

    2016-01-01

    The behaviour of two optical instruments, scilicet a laser scanning confocal microscope and a focus-variation microscope, was investigated considering measurements of tilted surfaces. The measured samples were twelve steel artefacts for mould surface finish reference, covering Sa roughness...... parameter in the range (101—103) nm. The 3D surface texture parameters considered were Sa, Sq and Sdq. The small working distance of the confocal microscope objectives influenced the measurement setup, preventing from selecting a high tilting angle. The investigation was carried out comparing measurements...... of flat surfaces (0° tilt) with measurements of 12.5° tilted surfaces. The confocal microscope results showed a high sensitivity to tilting due to the laser beam reflection on the metal surfaces. The focus variation microscope results were more robust with respect to the considered angular variation...

  10. Functional and ophthalmoscopic observations in human laser accident cases using scanning-laser ophthalmoscopy

    Science.gov (United States)

    Zwick, Harry; Lund, David J.; Gagliano, Donald A.; Stuck, Bruce E.

    1994-06-01

    A scanning laser ophthalmoscope (SLO) equipped with an acousto- optical modulator (ACM) was used to make focal acuity and contrast sensitivity measurements in individuals with macular damage. The depth of modulation achieved by the ACM was determined by imaging the SLO raster pattern onto a Pulnix TM 745 video camera and evaluating the intensity distribution with a Big Sky BVA10 beam view analyzer. Contrast levels remained approximately constant over the entire range of SLO input raster power settings. A delta Technologies image processing system produced Landolt ring test stimuli at the center of the raster pattern. Contrast thresholds were determined at various retinal locations by having subjects fixate a specific location on a fixed grid imaged on the raster pattern. This procedure insured that the test stimuli were always imaged in the center of the raster pattern thereby avoiding peripheral variations in the raster pattern intensity distribution. Measurements of contrast sensitivity where focal test targets fell within the macular damage area demonstrated elevated contrast thresholds relative to retinal locations where focal test targets evaluated the border regions between normal and pathological retina.

  11. A low error reconstruction method for confocal holography to determine 3-dimensional properties

    Energy Technology Data Exchange (ETDEWEB)

    Jacquemin, P.B., E-mail: pbjacque@nps.edu [Mechanical Engineering, University of Victoria, EOW 548,800 Finnerty Road, Victoria, BC (Canada); Herring, R.A. [Mechanical Engineering, University of Victoria, EOW 548,800 Finnerty Road, Victoria, BC (Canada)

    2012-06-15

    A confocal holography microscope developed at the University of Victoria uniquely combines holography with a scanning confocal microscope to non-intrusively measure fluid temperatures in three-dimensions (Herring, 1997), (Abe and Iwasaki, 1999), (Jacquemin et al., 2005). The Confocal Scanning Laser Holography (CSLH) microscope was built and tested to verify the concept of 3D temperature reconstruction from scanned holograms. The CSLH microscope used a focused laser to non-intrusively probe a heated fluid specimen. The focused beam probed the specimen instead of a collimated beam in order to obtain different phase-shift data for each scan position. A collimated beam produced the same information for scanning along the optical propagation z-axis. No rotational scanning mechanisms were used in the CSLH microscope which restricted the scan angle to the cone angle of the probe beam. Limited viewing angle scanning from a single view point window produced a challenge for tomographic 3D reconstruction. The reconstruction matrices were either singular or ill-conditioned making reconstruction with significant error or impossible. Establishing boundary conditions with a particular scanning geometry resulted in a method of reconstruction with low error referred to as 'wily'. The wily reconstruction method can be applied to microscopy situations requiring 3D imaging where there is a single viewpoint window, a probe beam with high numerical aperture, and specified boundary conditions for the specimen. The issues and progress of the wily algorithm for the CSLH microscope are reported herein. -- Highlights: Black-Right-Pointing-Pointer Evaluation of an optical confocal holography device to measure 3D temperature of a heated fluid. Black-Right-Pointing-Pointer Processing of multiple holograms containing the cumulative refractive index through the fluid. Black-Right-Pointing-Pointer Reconstruction issues due to restricting angular scanning to the numerical aperture of the

  12. A low error reconstruction method for confocal holography to determine 3-dimensional properties

    International Nuclear Information System (INIS)

    Jacquemin, P.B.; Herring, R.A.

    2012-01-01

    A confocal holography microscope developed at the University of Victoria uniquely combines holography with a scanning confocal microscope to non-intrusively measure fluid temperatures in three-dimensions (Herring, 1997), (Abe and Iwasaki, 1999), (Jacquemin et al., 2005). The Confocal Scanning Laser Holography (CSLH) microscope was built and tested to verify the concept of 3D temperature reconstruction from scanned holograms. The CSLH microscope used a focused laser to non-intrusively probe a heated fluid specimen. The focused beam probed the specimen instead of a collimated beam in order to obtain different phase-shift data for each scan position. A collimated beam produced the same information for scanning along the optical propagation z-axis. No rotational scanning mechanisms were used in the CSLH microscope which restricted the scan angle to the cone angle of the probe beam. Limited viewing angle scanning from a single view point window produced a challenge for tomographic 3D reconstruction. The reconstruction matrices were either singular or ill-conditioned making reconstruction with significant error or impossible. Establishing boundary conditions with a particular scanning geometry resulted in a method of reconstruction with low error referred to as “wily”. The wily reconstruction method can be applied to microscopy situations requiring 3D imaging where there is a single viewpoint window, a probe beam with high numerical aperture, and specified boundary conditions for the specimen. The issues and progress of the wily algorithm for the CSLH microscope are reported herein. -- Highlights: ► Evaluation of an optical confocal holography device to measure 3D temperature of a heated fluid. ► Processing of multiple holograms containing the cumulative refractive index through the fluid. ► Reconstruction issues due to restricting angular scanning to the numerical aperture of the beam. ► Minimizing tomographic reconstruction error by defining boundary

  13. Scanning vs. single spot laser ablation (λ=213 nm) inductively coupled plasma mass spectrometry

    International Nuclear Information System (INIS)

    Gonzalez, Jhanis J.; Fernandez, Alberto; Mao Xianglei; Russo, Richard E.

    2004-01-01

    Sampling strategy is defined in this work as the interaction of a repetitively pulsed laser beam with a fixed position on a sample (single spot) or with a moving sample (scan). Analytical performance of these sampling strategies was compared by using 213 nm laser ablation ICP-MS. A geological rock (Tuff) was quantitatively analyzed based on NIST series 610-616 glass standard reference materials. Laser ablation data were compared to ICP-MS analysis of the dissolved samples. The scan strategy (50 μm/s) produced a flat, steady temporal ICP-MS response whereas the single spot strategy produced a signal that decayed with time (after 60 s). Single-spot sampling provided better accuracy and precision than the scan strategy when the first 15 s of the sampling time was eliminated from the data analysis. In addition, the single spot strategy showed less matrix dependence among the four NIST glasses

  14. Il laser scanning e CloudCUBE per le grotte di Naica

    Directory of Open Access Journals (Sweden)

    Erminio Paolo Canevese

    2008-03-01

    Full Text Available Laser scanning and CloudCube for Naica caves On May 2007, Virtualgeo, a geomatic software development and communication company, took part in the first official expedition to Mexico. The Project, coined "Naica", involves researchers from ten universities, four companies and several laboratories. Virtualgeo carried out the survey by applying laser scanning technology to hypogeal caves covered with selenite crystals. The data was processed using CloudCUBE, a proprietary software designed to manage and model 3D point clouds. The first results of the laser scanning survey of a spectacular “forest of crystals” are presented here.

  15. Il laser scanning e CloudCUBE per le grotte di Naica

    Directory of Open Access Journals (Sweden)

    Erminio Paolo Canevese

    2008-03-01

    Full Text Available Laser scanning and CloudCube for Naica cavesOn May 2007, Virtualgeo, a geomatic software development and communication company, took part in the first official expedition to Mexico. The Project, coined "Naica", involves researchers from ten universities, four companies and several laboratories. Virtualgeo carried out the survey by applying laser scanning technology to hypogeal caves covered with selenite crystals. The data was processed using CloudCUBE, a proprietary software designed to manage and model 3D point clouds. The first results of the laser scanning survey of a spectacular “forest of crystals” are presented here.

  16. Rapid Prototyping — A Tool for Presenting 3-Dimensional Digital Models Produced by Terrestrial Laser Scanning

    Directory of Open Access Journals (Sweden)

    Juho-Pekka Virtanen

    2014-07-01

    Full Text Available Rapid prototyping has received considerable interest with the introduction of affordable rapid prototyping machines. These machines can be used to manufacture physical models from three-dimensional digital mesh models. In this paper, we compare the results obtained with a new, affordable, rapid prototyping machine, and a traditional professional machine. Two separate data sets are used for this, both of which were acquired using terrestrial laser scanning. Both of the machines were able to produce complex and highly detailed geometries in plastic material from models based on terrestrial laser scanning. The dimensional accuracies and detail levels of the machines were comparable, and the physical artifacts caused by the fused deposition modeling (FDM technique used in the rapid prototyping machines could be found in both models. The accuracy of terrestrial laser scanning exceeded the requirements for manufacturing physical models of large statues and building segments at a 1:40 scale.

  17. Automatic concrete cracks detection and mapping of terrestrial laser scan data

    Directory of Open Access Journals (Sweden)

    Mostafa Rabah

    2013-12-01

    The current paper submits a method for automatic concrete cracks detection and mapping from the data that was obtained during laser scanning survey. The method of cracks detection and mapping is achieved by three steps, namely the step of shading correction in the original image, step of crack detection and finally step of crack mapping and processing steps. The detected crack is defined in a pixel coordinate system. To remap the crack into the referred coordinate system, a reverse engineering is used. This is achieved by a hybrid concept of terrestrial laser-scanner point clouds and the corresponding camera image, i.e. a conversion from the pixel coordinate system to the terrestrial laser-scanner or global coordinate system. The results of the experiment show that the mean differences between terrestrial laser scan and the total station are about 30.5, 16.4 and 14.3 mms in x, y and z direction, respectively.

  18. Graph Structure-Based Simultaneous Localization and Mapping Using a Hybrid Method of 2D Laser Scan and Monocular Camera Image in Environments with Laser Scan Ambiguity

    Directory of Open Access Journals (Sweden)

    Taekjun Oh

    2015-07-01

    Full Text Available Localization is an essential issue for robot navigation, allowing the robot to perform tasks autonomously. However, in environments with laser scan ambiguity, such as long corridors, the conventional SLAM (simultaneous localization and mapping algorithms exploiting a laser scanner may not estimate the robot pose robustly. To resolve this problem, we propose a novel localization approach based on a hybrid method incorporating a 2D laser scanner and a monocular camera in the framework of a graph structure-based SLAM. 3D coordinates of image feature points are acquired through the hybrid method, with the assumption that the wall is normal to the ground and vertically flat. However, this assumption can be relieved, because the subsequent feature matching process rejects the outliers on an inclined or non-flat wall. Through graph optimization with constraints generated by the hybrid method, the final robot pose is estimated. To verify the effectiveness of the proposed method, real experiments were conducted in an indoor environment with a long corridor. The experimental results were compared with those of the conventional GMappingapproach. The results demonstrate that it is possible to localize the robot in environments with laser scan ambiguity in real time, and the performance of the proposed method is superior to that of the conventional approach.

  19. Centimeter-scale MEMS scanning mirrors for high power laser application

    Science.gov (United States)

    Senger, F.; Hofmann, U.; v. Wantoch, T.; Mallas, C.; Janes, J.; Benecke, W.; Herwig, Patrick; Gawlitza, P.; Ortega-Delgado, M.; Grune, C.; Hannweber, J.; Wetzig, A.

    2015-02-01

    A higher achievable scan speed and the capability to integrate two scan axes in a very compact device are fundamental advantages of MEMS scanning mirrors over conventional galvanometric scanners. There is a growing demand for biaxial high speed scanning systems complementing the rapid progress of high power lasers for enabling the development of new high throughput manufacturing processes. This paper presents concept, design, fabrication and test of biaxial large aperture MEMS scanning mirrors (LAMM) with aperture sizes up to 20 mm for use in high-power laser applications. To keep static and dynamic deformation of the mirror acceptably low all MEMS mirrors exhibit full substrate thickness of 725 μm. The LAMM-scanners are being vacuum packaged on wafer-level based on a stack of 4 wafers. Scanners with aperture sizes up to 12 mm are designed as a 4-DOF-oscillator with amplitude magnification applying electrostatic actuation for driving a motor-frame. As an example a 7-mm-scanner is presented that achieves an optical scan angle of 32 degrees at 3.2 kHz. LAMM-scanners with apertures sizes of 20 mm are designed as passive high-Q-resonators to be externally excited by low-cost electromagnetic or piezoelectric drives. Multi-layer dielectric coatings with a reflectivity higher than 99.9 % have enabled to apply cw-laser power loads of more than 600 W without damaging the MEMS mirror. Finally, a new excitation concept for resonant scanners is presented providing advantageous shaping of intensity profiles of projected laser patterns without modulating the laser. This is of interest in lighting applications such as automotive laser headlights.

  20. Investigation of laser cleaning on bronze cultural relics

    International Nuclear Information System (INIS)

    Ling, Xiulan; Wang, Gao; Zhang, Chen

    2016-01-01

    The effects of laser cleaning on the corrosion layers of bronze cultural relics were studied using a pulsed fiber laser. The laser cleaning threshold value of the corrosion layers was obtained. It was found that the corrosion layer was removed successfully by employing a laser fluence value of 0.32 J cm −2 and scanning for three times. To obtain experimental evidence, laser con-focal scanning microscopy, scanning electron microscopy (SEM), energy dispersive x-ray spectroscopy (EDS), laser induced breakdown spectroscopy (LIBS) and laser Raman spectroscopy were employed to investigate the cleaning efficiency of corrosion layers on specimens. (paper)

  1. Distribution of Biomolecules in Porous Nitrocellulose Membrane Pads Using Confocal Laser Scanning Microscopy and High-Speed Cameras

    NARCIS (Netherlands)

    Mujawar, Liyakat Hamid; Maan, Abid Aslam; Khan, Muhammad Kashif Iqbal; Norde, Willem; van Amerongen, Aart

    2013-01-01

    The main focus of our research was to study the distribution of inkjet printed biomolecules in porous nitrocellulose membrane pads of different brands. We produced microarrays of fluorophore-labeled IgG and bovine serum albumin (BSA) on FAST, Unisart, and Oncyte-Avid slides and compared the spot

  2. QUANTITATIVE FLUORESCENCE OF 5-FU-TREATED FETAL RAT LIMBS USING CONFOCAL LASER SCANNING MICROSCOPY AND LYSOTRACKER RED

    Science.gov (United States)

    Background: LysoTracker Red (LT) is a paraformaldehyde fixable probe that concentrates into acidic compartments of cells and tissues. After cell death a high level of lysosomal activity (acidic enzyme) is expressed resulting from phagocytosis of apoptotic bodies by neighboring ce...

  3. Extrusion and intrusion evolution in cyclically strained cast superalloy Inconel 738LC using confocal laser scanning microscope and AFM

    Czech Academy of Sciences Publication Activity Database

    Obrtlík, Karel; Juliš, M.; Man, Jiří; Podrábský, T.; Polák, Jaroslav

    2010-01-01

    Roč. 241, - (2010), Art. No. 012054 ISSN 1742-6588. [ICSMA-15 (15th International Conference on the Strength of Materials). Dresden, 16.08.2009-21.08.2009] R&D Projects: GA AV ČR 1QS200410502; GA ČR GA106/07/1507 Institutional research plan: CEZ:AV0Z20410507 Keywords : fatigue * persistent slip marking * Inconel 738LC Subject RIV: JL - Materials Fatigue, Friction Mechanics http://iopscience.iop.org/1742-6596/240/1/012054

  4. Confocal Raman Microscopy; applications in tissue engineering

    NARCIS (Netherlands)

    van Apeldoorn, Aart A.

    2005-01-01

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

  5. [Application Progress of Three-dimensional Laser Scanning Technology in Medical Surface Mapping].

    Science.gov (United States)

    Zhang, Yonghong; Hou, He; Han, Yuchuan; Wang, Ning; Zhang, Ying; Zhu, Xianfeng; Wang, Mingshi

    2016-04-01

    The booming three-dimensional laser scanning technology can efficiently and effectively get spatial three-dimensional coordinates of the detected object surface and reconstruct the image at high speed,high precision and large capacity of information.Non-radiation,non-contact and the ability of visualization make it increasingly popular in three-dimensional surface medical mapping.This paper reviews the applications and developments of three-dimensional laser scanning technology in medical field,especially in stomatology,plastic surgery and orthopedics.Furthermore,the paper also discusses the application prospects in the future as well as the biomedical engineering problems it would encounter with.

  6. D Model of AL Zubarah Fortress in Qatar - Terrestrial Laser Scanning VS. Dense Image Matching

    Science.gov (United States)

    Kersten, T.; Mechelke, K.; Maziull, L.

    2015-02-01

    In September 2011 the fortress Al Zubarah, built in 1938 as a typical Arabic fortress and restored in 1987 as a museum, was recorded by the HafenCity University Hamburg using terrestrial laser scanning with the IMAGER 5006h and digital photogrammetry for the Qatar Museum Authority within the framework of the Qatar Islamic Archaeology and Heritage Project. One goal of the object recording was to provide detailed 2D/3D documentation of the fortress. This was used to complete specific detailed restoration work in the recent years. From the registered laser scanning point clouds several cuttings and 2D plans were generated as well as a 3D surface model by triangle meshing. Additionally, point clouds and surface models were automatically generated from digital imagery from a Nikon D70 using the open-source software Bundler/PMVS2, free software VisualSFM, Autodesk Web Service 123D Catch beta, and low-cost software Agisoft PhotoScan. These outputs were compared with the results from terrestrial laser scanning. The point clouds and surface models derived from imagery could not achieve the same quality of geometrical accuracy as laser scanning (i.e. 1-2 cm).

  7. MULTISPECTRAL AIRBORNE LASER SCANNING - A NEW TREND IN THE DEVELOPMENT OF LIDAR TECHNOLOGY

    Directory of Open Access Journals (Sweden)

    Bakuła Krzysztof

    2015-12-01

    Full Text Available Airborne laser scanning (ALS is the one of the most accurate remote sensing techniques for data acquisition where the terrain and its coverage is concerned. Modern scanners have been able to scan in two or more channels (frequencies of the laser recently. This gives the rise to the possibility of obtaining diverse information about an area with the different spectral properties of objects. The paper presents an example of a multispectral ALS system - Titan by Optech - with the possibility of data including the analysis of digital elevation models accuracy and data density. As a result of the study, the high relative accuracy of LiDAR acquisition in three spectral bands was proven. The mean differences between digital terrain models (DTMs were less than 0.03 m. The data density analysis showed the influence of the laser wavelength. The points clouds that were tested had average densities of 25, 23 and 20 points per square metre respectively for green (G, near-infrared (NIR and shortwave-infrared (SWIR lasers. In this paper, the possibility of the generation of colour composites using orthoimages of laser intensity reflectance and its classification capabilities using data from airborne multispectral laser scanning for land cover mapping are also discussed and compared with conventional photogrammetric techniques.

  8. A flexible 3D laser scanning system using a robotic arm

    Science.gov (United States)

    Fei, Zixuan; Zhou, Xiang; Gao, Xiaofei; Zhang, Guanliang

    2017-06-01

    In this paper, we present a flexible 3D scanning system based on a MEMS scanner mounted on an industrial arm with a turntable. This system has 7-degrees of freedom and is able to conduct a full field scan from any angle, suitable for scanning object with the complex shape. The existing non-contact 3D scanning system usually uses laser scanner that projects fixed stripe mounted on the Coordinate Measuring Machine (CMM) or industrial robot. These existing systems can't perform path planning without CAD models. The 3D scanning system presented in this paper can scan the object without CAD models, and we introduced this path planning method in the paper. We also propose a practical approach to calibrating the hand-in-eye system based on binocular stereo vision and analyzes the errors of the hand-eye calibration.

  9. Remote defect imaging for plate-like structures based on the scanning laser source technique

    Science.gov (United States)

    Hayashi, Takahiro; Maeda, Atsuya; Nakao, Shogo

    2018-04-01

    In defect imaging with a scanning laser source technique, the use of a fixed receiver realizes stable measurements of flexural waves generated by laser at multiple rastering points. This study discussed the defect imaging by remote measurements using a laser Doppler vibrometer as a receiver. Narrow-band burst waves were generated by modulating laser pulse trains of a fiber laser to enhance signal to noise ratio in frequency domain. Averaging three images obtained at three different frequencies suppressed spurious distributions due to resonance. The experimental system equipped with these newly-devised means enabled us to visualize defects and adhesive objects in plate-like structures such as a plate with complex geometries and a branch pipe.

  10. Validation of a new noniterative method for accurate position determination of a scanning laser vibrometer

    Science.gov (United States)

    Pauwels, Steven; Boucart, Nick; Dierckx, Benoit; Van Vlierberghe, Pieter

    2000-05-01

    The use of a scanning laser Doppler vibrometer for vibration testing is becoming a popular instrument. The scanning laser Doppler vibrometer is a non-contacting transducer that can measure many points at a high spatial resolution in a short time. Manually aiming the laser beam at the points that need to be measured is very time consuming. In order to use it effectively, the position of the laser Doppler vibrometer needs to be determined relative to the structure. If the position of the laser Doppler vibrometer is known, any visible point on the structure can be hit and measured automatically. A new algorithm for this position determination is developed, based on a geometry model of the structure. After manually aiming the laser beam at 4 or more known points, the laser position and orientation relative to the structure is determined. Using this calculated position and orientation a list with the mirror angles for every measurement point is generated, which is used during the measurement. The algorithm is validated using 3 practical cases. In the first case a plate is used of which the points are measured very accurately, so the geometry model is assumed to be perfect. The second case is a brake disc. Here the geometry points are measured with a ruler, thus not so accurate. The final validation is done on a body in white of a car. A reduced finite element model is used as geometry model. This calibration shows that the new algorithm is very effective and practically usable.

  11. Efficient green lasers for high-resolution scanning micro-projector displays

    Science.gov (United States)

    Bhatia, Vikram; Bauco, Anthony S.; Oubei, Hassan M.; Loeber, David A. S.

    2010-02-01

    Laser-based projectors are gaining increased acceptance in mobile device market due to their low power consumption, superior image quality and small size. The basic configuration of such micro-projectors is a miniature mirror that creates an image by raster scanning the collinear red, blue and green laser beams that are individually modulated on a pixel-bypixel basis. The image resolution of these displays can be limited by the modulation bandwidth of the laser sources, and the modulation speed of the green laser has been one of the key limitations in the development of these displays. We will discuss how this limitation is fundamental to the architecture of many laser designs and then present a green laser configuration which overcomes these difficulties. In this green laser architecture infra-red light from a distributed Bragg-reflector (DBR) laser diode undergoes conversion to green light in a waveguided second harmonic generator (SHG) crystal. The direct doubling in a single pass through the SHG crystal allows the device to operate at the large modulation bandwidth of the DBR laser. We demonstrate that the resultant product has a small footprint (9% electrical-to-optical conversion) and large modulation bandwidth (>100 MHz).

  12. Topographic laser ranging and scanning principles and processing

    CERN Document Server

    Shan, Jie

    2008-01-01

    A systematic, in-depth introduction to theories and principles of Light Detection and Ranging (LiDAR) technology is long overdue, as it is the most important geospatial data acquisition technology to be introduced in recent years. An advanced discussion, this text fills the void.Professionals in fields ranging from geology, geography and geoinformatics to physics, transportation, and law enforcement will benefit from this comprehensive discussion of topographic LiDAR principles, systems, data acquisition, and data processing techniques. The book covers ranging and scanning fundamentals, and broad, contemporary analysis of airborne LiDAR systems, as well as those situated on land and in space. The authors present data collection at the signal level in terms of waveforms and their properties; at the system level with regard to calibration and georeferencing; and at the data level to discuss error budget, quality control, and data organization. They devote the bulk of the book to LiDAR data processing and inform...

  13. Confocal Laser Endomicroscopy for the Diagnosis of Urothelial Carcinoma in the Bladder and the Upper Urinary Tract: Protocols for Two Prospective Explorative Studies.

    Science.gov (United States)

    Liem, Esmee Iml; Freund, Jan Erik; Baard, Joyce; de Bruin, D Martijn; Laguna Pes, M Pilar; Savci-Heijink, C Dilara; van Leeuwen, Ton G; de Reijke, Theo M; de la Rosette, Jean Jmch

    2018-02-07

    Visual confirmation of a suspicious lesion in the urinary tract is a major corner stone in diagnosing urothelial carcinoma. However, during cystoscopy (for bladder tumors) and ureterorenoscopy (for tumors of the upper urinary tract) no real-time histopathologic information can be obtained. Confocal laser endomicroscopy (CLE) is an optical imaging technique that allows for in vivo high-resolution imaging and may allow real-time tumor grading of urothelial lesions. The primary objective of both studies is to develop descriptive criteria for in vivo CLE images of urothelial carcinoma (low-grade, high-grade, carcinoma in situ) and normal urothelium by comparing CLE images with corresponding histopathology. In these two prospective clinical trials, CLE imaging will be performed of suspicious lesions and normal tissue in the urinary tract during surgery, prior to resection or biopsy. In the bladder study, CLE will be performed in 60 patients using the Cystoflex UHD-R probe. In the upper urinary tract study, CLE will be performed in 25 patients during ureterorenoscopy, who will undergo radical treatment (nephroureterectomy or segmental ureter resection) thereafter. All CLE images will be analyzed frame by frame by three independent, blinded observers. Histopathology and CLE-based diagnosis of the lesions will be evaluated. Both studies comply with the IDEAL stage 2b recommendations. Presently, recruitment of patients is ongoing in both studies. Results and outcomes are expected in 2018. For development of CLE-based diagnosis of urothelial carcinoma in the bladder and the upper urinary tract, a structured conduct of research is required. This study will provide more insight in tissue-specific CLE criteria for real-time tumor grading of urothelial carcinoma. Confocal Laser Endomicroscopy: ClinicalTrials.gov NCT03013894; https://clinicaltrials.gov /ct2/show/NCT03013894?term=NCT03013894&rank=1 (Archived by WebCite at http://www.webcitation.org/6wiPZ378I); and Dutch Central

  14. Detection of fluorescent organic nanoparticles by confocal laser endomicroscopy in a rat model of Barrett’s esophageal adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Dassie E

    2015-10-01

    . Non-operated and operated rats in which gastroesophageal reflux was surgically induced received both types of NPs (NP-DCM and NP-DCM-ASYNYDA by intravenous route. Localization of mucosal NPs was assessed in vivo by confocal laser endomicroscopy, a technique which enables a “real time” and in situ visualization of the tissue at a cellular level. After injection of NP-DCM and NP-DCM-ASYNYDA, fluorescence was observed in rats affected by esophageal cancer, whereas no signal was observed in control non-operated rats, or in rats with simple esophagitis or Barrett’s esophagus mucosa. Fluorescence was observable in vivo 30 minutes after the administration of NPs. Interestingly, NP-DCM-ASYNYDA induced strong fluorescence intensity 24 hours after administration. These observations suggested that NPs could reach the tumor cells, likely by enhanced permeability and retention effect, and the peptide ASYNYDA gave them high specificity for esophageal cancer cells. Thus, the combination of NP platform and confocal laser endomicroscopy could play an important role for highlighting esophageal cancer conditions. This result supports the potential of this strategy as a targeted carrier for photoactive and bioactive molecules in esophageal cancer diagnosis and treatment. Keywords: confocal laser endomicroscopy, Barrett’s esophagus, diagnostics, esophageal adenocarcinoma, fluorescent nanoparticles, heptapeptide

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

    Science.gov (United States)

    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.

  16. Fan-beam scanning laser optical computed tomography for large volume dosimetry

    Science.gov (United States)

    Dekker, K. H.; Battista, J. J.; Jordan, K. J.

    2017-05-01

    A prototype scanning-laser fan beam optical CT scanner is reported which is capable of high resolution, large volume dosimetry with reasonable scan time. An acylindrical, asymmetric aquarium design is presented which serves to 1) generate parallel-beam scan geometry, 2) focus light towards a small acceptance angle detector, and 3) avoid interference fringe-related artifacts. Preliminary experiments with uniform solution phantoms (11 and 15 cm diameter) and finger phantoms (13.5 mm diameter FEP tubing) demonstrate that the design allows accurate optical CT imaging, with optical CT measurements agreeing within 3% of independent Beer-Lambert law calculations.

  17. Fan-beam scanning laser optical computed tomography for large volume dosimetry

    International Nuclear Information System (INIS)

    Dekker, K H; Battista, J J; Jordan, K J

    2017-01-01

    A prototype scanning-laser fan beam optical CT scanner is reported which is capable of high resolution, large volume dosimetry with reasonable scan time. An acylindrical, asymmetric aquarium design is presented which serves to 1) generate parallel-beam scan geometry, 2) focus light towards a small acceptance angle detector, and 3) avoid interference fringe-related artifacts. Preliminary experiments with uniform solution phantoms (11 and 15 cm diameter) and finger phantoms (13.5 mm diameter FEP tubing) demonstrate that the design allows accurate optical CT imaging, with optical CT measurements agreeing within 3% of independent Beer-Lambert law calculations. (paper)

  18. Glacier Snowline Determination from Terrestrial Laser Scanning Intensity Data

    Directory of Open Access Journals (Sweden)

    Hannah Prantl

    2017-07-01

    Full Text Available Accurately identifying the extent of surface snow cover on glaciers is important for extrapolating end of year mass balance measurements, constraining the glacier surface radiative energy balance and evaluating model simulations of snow cover. Here, we use auxiliary information from Riegl VZ-6000 Terrestrial Laser Scanner (TLS return signals to accurately map the snow cover over a glacier throughout an ablation season. Three classification systems were compared, and we find that supervised classification based on TLS signal intensity alone is outperformed by a rule-based classification employing intensity, surface roughness and an associated optical image, which achieves classification accuracy of 68–100%. The TLS intensity signal shows no meaningful relationship with surface or bulk snow density. Finally, we have also compared our Snow Line Altitude (SLA derived from TLS with SLA derived from the model output, as well as one Landsat image. The results of the model output track the SLA from TLS well, however with a positive bias. In contrast, automatic Landsat-derived SLA slightly underestimates the SLA from TLS. To conclude, we demonstrate that the snow cover extent can be mapped successfully using TLS, although the snow mass remains elusive.

  19. Application of 3D Laser Scanning Technology in Complex Rock Foundation Design

    Science.gov (United States)

    Junjie, Ma; Dan, Lu; Zhilong, Liu

    2017-12-01

    Taking the complex landform of Tanxi Mountain Landscape Bridge as an example, the application of 3D laser scanning technology in the mapping of complex rock foundations is studied in this paper. A set of 3D laser scanning technologies are formed and several key engineering problems are solved. The first is 3D laser scanning technology of complex landforms. 3D laser scanning technology is used to obtain a complete 3D point cloud data model of the complex landform. The detailed and accurate results of the surveying and mapping decrease the measuring time and supplementary measuring times. The second is 3D collaborative modeling of the complex landform. A 3D model of the complex landform is established based on the 3D point cloud data model. The super-structural foundation model is introduced for 3D collaborative design. The optimal design plan is selected and the construction progress is accelerated. And the last is finite-element analysis technology of the complex landform foundation. A 3D model of the complex landform is introduced into ANSYS for building a finite element model to calculate anti-slide stability of the rock, and provides a basis for the landform foundation design and construction.

  20. Real-time depth monitoring and control of laser machining through scanning beam delivery system

    International Nuclear Information System (INIS)

    Ji, Yang; Grindal, Alexander W; Fraser, James M; Webster, Paul J L

    2015-01-01

    Scanning optics enable many laser applications in manufacturing because their low inertia allows rapid movement of the process beam across the sample. We describe our method of inline coherent imaging for real-time (up to 230 kHz) micron-scale (7–8 µm axial resolution) tracking and control of laser machining depth through a scanning galvo-telecentric beam delivery system. For 1 cm trench etching in stainless steel, we collect high speed intrapulse and interpulse morphology which is useful for further understanding underlying mechanisms or comparison with numerical models. We also collect overall sweep-to-sweep depth penetration which can be used for feedback depth control. For trench etching in silicon, we show the relationship of etch rate with average power and scan speed by computer processing of depth information without destructive sample post-processing. We also achieve three-dimensional infrared continuous wave (modulated) laser machining of a 3.96 × 3.96 × 0.5 mm 3 (length × width × maximum depth) pattern on steel with depth feedback. To the best of our knowledge, this is the first successful demonstration of direct real-time depth monitoring and control of laser machining with scanning optics. (paper)

  1. A STUDY ABOUT TERRESTRIAL LASER SCANNING FOR RECONSTRUCTION OF PRECAST CONCRETE TO SUPPORT QLASSIC ASSESSMENT

    Directory of Open Access Journals (Sweden)

    M. A. Aziz

    2016-09-01

    Full Text Available Nowadays, terrestrial laser scanning shows the potential to improve construction productivity by measuring the objects changes using real-time applications. This paper presents the process of implementation of an efficient framework for precast concrete using terrestrial laser scanning that enables contractors to acquire accurate data and support Quality Assessment System in Construction (QLASSIC. Leica Scanstation C10, black/white target, Autodesk Revit and Cyclone software were used in this study. The results were compared with the dimensional of based model precast concrete given by the company as a reference with the AutoDesk Revit model from the terrestrial laser scanning data and conventional method (measuring tape. To support QLASSIC, the tolerance dimensions of cast in-situ & precast elements is +10mm / -5mm. The results showed that the root mean square error for a Revit model is 2.972mm while using measuring tape is 13.687mm. The accuracy showed that terrestrial laser scanning has an advantage in construction jobs to support QLASSIC.

  2. INITIAL TESTS AND ACCURACY ASSESMENT OF A COMPACT MOBILE LASER SCANNING SYSTEM

    Directory of Open Access Journals (Sweden)

    K. Julge

    2016-06-01

    Full Text Available Mobile laser scanning (MLS is a faster and cost-effective alternative to static laser scanning, even though there is a slight trade-off in accuracy. This contribution describes a compact mobile laser scanning system mounted on a vehicle. The technical parameters of the used system components, i.e. a small LIDAR sensor Velodyne VLP-16 and a dual antenna GNSS/INS system Advanced Navigation Spatial Dual, are reviewed, along with the integration of these components for spatial data acquisition. Calculation principles of 3D coordinates from the real-time data of all the involved sensors are discussed. The field tests were carried out in a controlled environment of a parking lot and at different velocities. Experiments were carried out to test the ability of the GNSS/INS system to cope with difficult conditions, e.g. sudden movements due to cornering or swerving. The accuracy of the resulting MLS point cloud is evaluated with respect to high-accuracy static terrestrial laser scanning data. Problems regarding combining LIDAR, GNSS and INS sensors are outlined, as well as the initial accuracy assessments. Initial tests revealed errors related to insufficient quality of inertial data and a need for the trajectory post-processing calculations. Although this study was carried out while the system was mounted on a car, there is potential for operating the system on an unmanned aerial vehicle, all-terrain vehicle or in a backpack mode due to its relatively compact size.

  3. Better Visualisation of Air-borne Laser Scanning for geomorphological and archaeological interpretation

    DEFF Research Database (Denmark)

    Ljungberg, Thomas; Scott, D; Kristiansen, Søren Munch

    Digital elevation models derived from high-precision Air-borne Laser Scanning (ALS or LiDAR) point clouds are becoming increasingly available throughout the world. These elevation models presents a very valuable tool for locating and interpreting geomorphological as well as archaeological features...

  4. Method for quantifying percentage wood failure in block-shear specimens by a laser scanning profilometer

    Science.gov (United States)

    C. T. Scott; R. Hernandez; C. Frihart; R. Gleisner; T. Tice

    2005-01-01

    A new method for quantifying percentage wood failure of an adhesively bonded block-shear specimen has been developed. This method incorporates a laser displacement gage with an automated two-axis positioning system that functions as a highly sensitive profilometer. The failed specimen is continuously scanned across its width to obtain a surface failure profile. The...

  5. An evaluation of the efficiency of laser scanning technology in the ...

    African Journals Online (AJOL)

    Green-Blue (RGB) intensity values for each point. Point clouds of data can now be imported into a CAD package and compared to design specifications. In the case where “as-built” specifications differ for the initial design, laser scanning allows ...

  6. Initial Tests and Accuracy Assesment of a Compact Mobile Laser Scanning System

    Science.gov (United States)

    Julge, K.; Ellmann, A.; Vajakas, T.; Kolka, R.

    2016-06-01

    Mobile laser scanning (MLS) is a faster and cost-effective alternative to static laser scanning, even though there is a slight trade-off in accuracy. This contribution describes a compact mobile laser scanning system mounted on a vehicle. The technical parameters of the used system components, i.e. a small LIDAR sensor Velodyne VLP-16 and a dual antenna GNSS/INS system Advanced Navigation Spatial Dual, are reviewed, along with the integration of these components for spatial data acquisition. Calculation principles of 3D coordinates from the real-time data of all the involved sensors are discussed. The field tests were carried out in a controlled environment of a parking lot and at different velocities. Experiments were carried out to test the ability of the GNSS/INS system to cope with difficult conditions, e.g. sudden movements due to cornering or swerving. The accuracy of the resulting MLS point cloud is evaluated with respect to high-accuracy static terrestrial laser scanning data. Problems regarding combining LIDAR, GNSS and INS sensors are outlined, as well as the initial accuracy assessments. Initial tests revealed errors related to insufficient quality of inertial data and a need for the trajectory post-processing calculations. Although this study was carried out while the system was mounted on a car, there is potential for operating the system on an unmanned aerial vehicle, all-terrain vehicle or in a backpack mode due to its relatively compact size.

  7. Laser Scanning Technology as Part of a Comprehensive Condition Assessment for Covered Bridges

    Science.gov (United States)

    Brian K. Brashaw; Samuel Anderson; Robert J. Ross

    2015-01-01

    New noncontact technologies have been developed and implemented for determining as-built condition and current dimensions for a wide variety of objects and buildings. In this study, a three-dimensional laser scanner was used to determine the dimensions and visual condition of a historic bridge in the Amnicon Falls State Park in northern Wisconsin. 3D scanning provides...

  8. Light propagation studies on laser modified waveguides using scanning near-field optical microscopy

    DEFF Research Database (Denmark)

    Borrise, X.; Berini, Abadal Gabriel; Jimenez, D.

    2001-01-01

    By means of direct laser writing on Al, a new method to locally modify optical waveguides is proposed. This technique has been applied to silicon nitride waveguides, allowing modifications of the optical propagation along the guide. To study the formed structures, a scanning near-held optical mic...

  9. a Study about Terrestrial Laser Scanning for Reconstruction of Precast Concrete to Support Qlassic Assessment

    Science.gov (United States)

    Aziz, M. A.; Idris, K. M.; Majid, Z.; Ariff, M. F. M.; Yusoff, A. R.; Luh, L. C.; Abbas, M. A.; Chong, A. K.

    2016-09-01

    Nowadays, terrestrial laser scanning shows the potential to improve construction productivity by measuring the objects changes using real-time applications. This paper presents the process of implementation of an efficient framework for precast concrete using terrestrial laser scanning that enables contractors to acquire accurate data and support Quality Assessment System in Construction (QLASSIC). Leica Scanstation C10, black/white target, Autodesk Revit and Cyclone software were used in this study. The results were compared with the dimensional of based model precast concrete given by the company as a reference with the AutoDesk Revit model from the terrestrial laser scanning data and conventional method (measuring tape). To support QLASSIC, the tolerance dimensions of cast in-situ & precast elements is +10mm / -5mm. The results showed that the root mean square error for a Revit model is 2.972mm while using measuring tape is 13.687mm. The accuracy showed that terrestrial laser scanning has an advantage in construction jobs to support QLASSIC.

  10. Multimodal scanning laser ophthalmoscopy for image guided treatment of age-related macular degeneration

    Science.gov (United States)

    Hammer, Daniel X.; Ferguson, R. D.; Patel, Ankit H.; Iftimia, Nicusor V.; Mujat, Mircea; Husain, Deeba

    2009-02-01

    Subretinal neovascular membranes (SRNM) are a deleterious complication of laser eye injury and retinal diseases such as age-related macular degeneration (AMD), choroiditis, and myopic retinopathy. Photodynamic therapy (PDT) and anti-vascular endothelial growth factor (VEGF) drugs are approved treatment methods. PDT acts by selective dye accumulation, activation by laser light, and disruption and clotting of the new leaky vessels. However, PDT surgery is currently not image-guided, nor does it proceed in an efficient or automated manner. This may contribute to the high rate of re-treatment. We have developed a multimodal scanning laser ophthalmoscope (SLO) for automated diagnosis and image-guided treatment of SRNMs associated with AMD. The system combines line scanning laser ophthalmoscopy (LSLO), fluorescein angiography (FA), indocyanine green angiography (ICGA), PDT laser delivery, and retinal tracking in a compact, efficient platform. This paper describes the system hardware and software design, performance characterization, and automated patient imaging and treatment session procedures and algorithms. Also, we present initial imaging and tracking measurements on normal subjects and automated lesion demarcation and sizing analysis of previously acquired angiograms. Future pre-clinical testing includes line scanning angiography and PDT treatment of AMD subjects. The automated acquisition procedure, enhanced and expedited data post-processing, and innovative image visualization and interpretation tools provided by the multimodal retinal imager may eventually aid in the diagnosis, treatment, and prognosis of AMD and other retinal diseases.

  11. ANALYSIS OF TERRESTRIAL LASER SCANNING AND PHOTOGRAMMETRY DATA FOR DOCUMENTATION OF HISTORICAL ARTIFACTS

    Directory of Open Access Journals (Sweden)

    R. A. Kuçak

    2016-10-01

    Full Text Available Historical artifacts living from the past until today exposed to many destructions non-naturally or naturally. For this reason, The protection and documentation studies of Cultural Heritage to inform the next generations are accelerating day by day in the whole world. The preservation of historical artifacts using advanced 3D measurement technologies becomes an efficient tool for mapping solutions. There are many methods for documentation and restoration of historic structures. In addition to traditional methods such as simple hand measurement and tachometry, terrestrial laser scanning is rapidly becoming one of the most commonly used techniques due to its completeness, accuracy and fastness characteristics. This study evaluates terrestrial laser scanning(TLS technology and photogrammetry for documenting the historical artifacts facade data in 3D Environment. PhotoModeler software developed by Eos System was preferred for Photogrammetric method. Leica HDS 6000 laser scanner developed by Leica Geosystems and Cyclone software which is the laser data evaluation software belonging to the company is preferred for Terrestrial Laser Scanning method. Taking into account the results obtained with this software product is intended to provide a contribution to the studies for the documentation of cultural heritage.

  12. Automatic Stem Mapping by Merging Several Terrestrial Laser Scans at the Feature and Decision Levels

    Directory of Open Access Journals (Sweden)

    Juha Hyyppä

    2013-01-01

    Full Text Available Detailed up-to-date ground reference data have become increasingly important in quantitative forest inventories. Field reference data are conventionally collected at the sample plot level by means of manual measurements, which are both labor-intensive and time-consuming. In addition, the number of attributes collected from the tree stem is limited. More recently, terrestrial laser scanning (TLS, using both single-scan and multi-scan techniques, has proven to be a promising solution for efficient stem mapping at the plot level. In the single-scan method, the laser scanner is placed at the center of the plot, creating only one scan, and all trees are mapped from the single-scan point cloud. Consequently, the occlusion of stems increases as the range of the scanner increases, depending on the forest’s attributes. In the conventional multi-scan method, several scans are made simultaneously inside and outside of the plot to collect point clouds representing all trees within the plot, and these scans are accurately co-registered by using artificial reference targets manually placed throughout the plot. The additional difficulty of applying the multi-scan method is due to the point-cloud registration of several scans not being fully automated yet. This paper proposes a multi-single-scan (MSS method to map the sample plot. The method does not require artificial reference targets placed on the plot or point-level registration. The MSS method is based on the fully automated processing of each scan independently and on the merging of the stem positions automatically detected from multiple scans to accurately map the sample plot. The proposed MSS method was tested on five dense forest plots. The results show that the MSS method significantly improves the stem-detection accuracy compared with the single-scan approach and achieves a mapping accuracy similar to that achieved with the multi-scan method, without the need for the point-level registration.

  13. Sensitivity and specificity of a new scoring system for diabetic macular oedema detection using a confocal laser imaging system

    Science.gov (United States)

    Tong, L; Ang, A; Vernon, S; Zambarakji, H; Bhan, A; Sung, V; Page, S

    2001-01-01

    AIM—To assess the use of the Heidelberg retina tomograph (HRT) in screening for sight threatening diabetic macular oedema in a hospital diabetic clinic, using a new subjective analysis system (SCORE).
METHODS—200 eyes of 100 consecutive diabetic patients attending a diabetologist's clinic were studied, all eyes had an acuity of 6/9 or better. All patients underwent clinical examination by an ophthalmologist. Using the HRT, one good scan was obtained for each eye centred on the fovea. A System for Classification and Ordering of Retinal Edema (SCORE) was developed using subjective assessment of the colour map and the reflectivity image. The interobserver agreement of using this method to detect macular oedema was assessed by two observers (ophthalmic trainees) who were familiarised with SCORE by studying standard pictures of eyes not in the study. All scans were graded from 0-6 and test positive cases were defined as having a SCORE value of 0-2. The sensitivity of SCORE was assessed by pooling the data with an additional 88 scans of 88 eyes in order to reduce the confidence interval of the index.
RESULTS—12 eyes in eight out of the 100 patients had macular oedema clinically. Three scans in three patients could not be analysed because of poor scan quality. In the additional group of scans 76 out of 88 eyes had macular oedema clinically. The scoring system had a specificity of 99% (95% CI 96-100) and sensitivity of 67% (95% CI 57-76). The predictive value of a negative test was 87% (95% CI 82-99), and that of a positive test was 95% (95% CI 86-99). The mean difference of the SCORE value between two observers was -0.2 (95% CI -0.5 to +0.07).
CONCLUSIONS—These data suggest that SCORE is potentially useful for detecting diabetic macular oedema in hospital diabetic patients.

 PMID:11133709

  14. Tritium Removal from Codeposits on Carbon Tiles by a Scanning Laser

    International Nuclear Information System (INIS)

    C.H. Skinner; C.A. Gentile; A. Carpe; G. Guttadora; S. Langish; K.M. Young; W.M. Shu; H. Nakamura

    2001-01-01

    A novel method for tritium release has been demonstrated on codeposited layers on graphite and carbon-fiber-composite tiles from the Tokamak Fusion Test Reactor (TFTR). A scanning continuous wave Nd laser beam heated the codeposits to a temperature of 1200-2300 degrees C for 10 to 200 milliseconds in an argon atmosphere. The temperature rise of the codeposit was significantly higher than that of the manufactured tile material (e.g., 1770 degrees C cf. 1080 degrees C). A major fraction of tritium was thermally desorbed with minimal change to the surface appearance at a laser intensity of 8 kW/cm(superscript ''2''), peak temperatures above 1230 degrees C and heating duration 10-20 milliseconds. In two experiments, 46% and 84% of the total tritium was released during the laser scan. The application of this method for tritium removal from a tokamak reactor appears promising and has significant advantages over oxidative techniques

  15. Forest Resource Measurements by Combination of Terrestrial Laser Scanning and Drone Use

    Science.gov (United States)

    Cheung, K.; Katoh, M.; Horisawa, M.

    2017-10-01

    Using terrestrial laser scanning (TLS), forest attributes such as diameter at breast height (DBH) and tree location can be measured accurately. However, due to low penetration of laser pulses to tree tops, tree height measurements are typically underestimated. In this study, data acquired by TLS and drones were combined; DBH and tree locations were determined by TLS, and tree heights were measured by drone use. The average tree height error and root mean square error (RMSE) of tree height were 0.8 and 1.2 m, respectively, for the combined method, and -0.4 and 1.7 m using TLS alone. The tree height difference was compared using airborne laser scanning (ALS). Furthermore, a method to acquire 100 % tree detection rate based on TLS data is suggested in this study.

  16. A novel near real-time laser scanning device for geometrical determination of pleural cavity surface.

    Science.gov (United States)

    Kim, Michele M; Zhu, Timothy C

    2013-02-02

    During HPPH-mediated pleural photodynamic therapy (PDT), it is critical to determine the anatomic geometry of the pleural surface quickly as there may be movement during treatment resulting in changes with the cavity. We have developed a laser scanning device for this purpose, which has the potential to obtain the surface geometry in real-time. A red diode laser with a holographic template to create a pattern and a camera with auto-focusing abilities are used to scan the cavity. In conjunction with a calibration with a known surface, we can use methods of triangulation to reconstruct the surface. Using a chest phantom, we are able to obtain a 360 degree scan of the interior in under 1 minute. The chest phantom scan was compared to an existing CT scan to determine its accuracy. The laser-camera separation can be determined through the calibration with 2mm accuracy. The device is best suited for environments that are on the scale of a chest cavity (between 10cm and 40cm). This technique has the potential to produce cavity geometry in real-time during treatment. This would enable PDT treatment dosage to be determined with greater accuracy. Works are ongoing to build a miniaturized device that moves the light source and camera via a fiber-optics bundle commonly used for endoscopy with increased accuracy.

  17. Error analysis of motion correction method for laser scanning of moving objects

    Science.gov (United States)

    Goel, S.; Lohani, B.

    2014-05-01

    The limitation of conventional laser scanning methods is that the objects being scanned should be static. The need of scanning moving objects has resulted in the development of new methods capable of generating correct 3D geometry of moving objects. Limited literature is available showing development of very few methods capable of catering to the problem of object motion during scanning. All the existing methods utilize their own models or sensors. Any studies on error modelling or analysis of any of the motion correction methods are found to be lacking in literature. In this paper, we develop the error budget and present the analysis of one such `motion correction' method. This method assumes availability of position and orientation information of the moving object which in general can be obtained by installing a POS system on board or by use of some tracking devices. It then uses this information along with laser scanner data to apply correction to laser data, thus resulting in correct geometry despite the object being mobile during scanning. The major application of this method lie in the shipping industry to scan ships either moving or parked in the sea and to scan other objects like hot air balloons or aerostats. It is to be noted that the other methods of "motion correction" explained in literature can not be applied to scan the objects mentioned here making the chosen method quite unique. This paper presents some interesting insights in to the functioning of "motion correction" method as well as a detailed account of the behavior and variation of the error due to different sensor components alone and in combination with each other. The analysis can be used to obtain insights in to optimal utilization of available components for achieving the best results.

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

    Science.gov (United States)

    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%.

  19. Automated inspection of gaps on the free-form shape parts by laser scanning technologies

    Science.gov (United States)

    Zhou, Sen; Xu, Jian; Tao, Lei; An, Lu; Yu, Yan

    2018-01-01

    In industrial manufacturing processes, the dimensional inspection of the gaps on the free-form shape parts is critical and challenging, and is directly associated with subsequent assembly and terminal product quality. In this paper, a fast measuring method for automated gap inspection based on laser scanning technologies is presented. The proposed measuring method consists of three steps: firstly, the relative position is determined according to the geometric feature of measuring gap, which considers constraints existing in a laser scanning operation. Secondly, in order to acquire a complete gap profile, a fast and effective scanning path is designed. Finally, the range dimension of the gaps on the free-form shape parts including width, depth and flush, correspondingly, is described in a virtual environment. In the future, an appliance machine based on the proposed method will be developed for the on-line dimensional inspection of gaps on the automobile or aerospace production line.

  20. In vitro confocal imaging of the rabbit cornea.

    Science.gov (United States)

    Masters, B R; Paddock, S

    1990-05-01

    We were able to observe in vitro the fine structure of the rabbit cornea using a laser scanning confocal microscope, especially in the regions between Descemet's membrane and the epithelial basal lamina. We observed submicrometre filaments throughout the stroma with high concentrations adjacent to Descemet's membrane, and found extensive interconnecting processes between stromal keratocytes. There are numerous regions containing nerve plexuses in the stroma. We found a deeply convoluted basal lamina adjacent to the epithelium, and observed regions containing junctions between endothelial cells in fluorescent images of rabbit corneas stained with the actin-specific compound fluorescein phalloidin.

  1. Capturing and modelling high-complex alluvial topography with UAS-borne laser scanning

    Science.gov (United States)

    Mandlburger, Gottfried; Wieser, Martin; Pfennigbauer, Martin

    2015-04-01

    Due to fluvial activity alluvial forests are zones of highest complexity and relief energy. Alluvial forests are dominated by new and pristine channels in consequence of current and historic flood events. Apart from topographic features, the vegetation structure is typically very complex featuring, both, dense under story as well as high trees. Furthermore, deadwood and debris carried from upstream during periods of high discharge within the river channel are deposited in these areas. Therefore, precise modelling of the micro relief of alluvial forests using standard tools like Airborne Laser Scanning (ALS) is hardly feasible. Terrestrial Laser Scanning (TLS), in turn, is very time consuming for capturing larger areas as many scan positions are necessary for obtaining complete coverage due to view occlusions in the forest. In the recent past, the technological development of Unmanned Arial Systems (UAS) has reached a level that light-weight survey-grade laser scanners can be operated from these platforms. For capturing alluvial topography this could bridge the gap between ALS and TLS in terms of providing a very detailed description of the topography and the vegetation structure due to the achievable very high point density of >100 points per m2. In our contribution we demonstrate the feasibility to apply UAS-borne laser scanning for capturing and modelling the complex topography of the study area Neubacher Au, an alluvial forest at the pre-alpine River Pielach (Lower Austria). The area was captured with Riegl's VUX-1 compact time-of-flight laser scanner mounted on a RiCopter (X-8 array octocopter). The scanner features an effective scan rate of 500 kHz and was flown in 50-100 m above ground. At this flying height the laser footprint is 25-50 mm allowing mapping of very small surface details. Furthermore, online waveform processing of the backscattered laser energy enables the retrieval of multiple targets for single laser shots resulting in a dense point cloud of

  2. Corneal Backscatter Analysis by In Vivo Confocal Microscopy: Fellow Eye Comparison of Small Incision Lenticule Extraction and Femtosecond Laser-Assisted LASIK

    Directory of Open Access Journals (Sweden)

    Alper Agca

    2014-01-01

    Full Text Available Purpose. To evaluate and compare corneal backscatter from anterior stroma between small incision lenticule extraction (SMILE and femtosecond laser-assisted LASIK (femto-LASIK. Methods. A cohort of 60 eyes of 30 patients was randomized to receive SMILE in one eye and femto-LASIK in the fellow eye. In vivo confocal microscopy was performed at 1 week and 1, 3, and 6 months after surgery. The main outcome measurements were maximum backscattered intensity and the depth from which it was measured, the backscattered light intensity 30 μm below Bowman’s membrane at the flap interface and 150 μm below the superficial epithelium, and the number of refractive particles at the flap interface. Results. The mean backscattered light intensity (LI at all measured depths and the maximum backscattered LI were higher in the SMILE group than the femto-LASIK group at all postoperative visits. LI differences at 1 week and 1- and 3-month visits were statistically significant (P<0,05. LI differences at 6 months were not statistically significant. There was no difference in the number of refractive particles at the flap interface between the groups at any visit. Conclusions. SMILE results in increased backscattered LI in the anterior stroma when compared with femto-LASIK were evaluated.

  3. Keypoint-based 4-Points Congruent Sets - Automated marker-less registration of laser scans

    Science.gov (United States)

    Theiler, Pascal Willy; Wegner, Jan Dirk; Schindler, Konrad

    2014-10-01

    We propose a method to automatically register two point clouds acquired with a terrestrial laser scanner without placing any markers in the scene. What makes this task challenging are the strongly varying point densities caused by the line-of-sight measurement principle, and the huge amount of data. The first property leads to low point densities in potential overlap areas with scans taken from different viewpoints while the latter calls for highly efficient methods in terms of runtime and memory requirements. A crucial yet largely unsolved step is the initial coarse alignment of two scans without any simplifying assumptions, that is, point clouds are given in arbitrary local coordinates and no knowledge about their relative orientation is available. Once coarse alignment has been solved, scans can easily be fine-registered with standard methods like least-squares surface or Iterative Closest Point matching. In order to drastically thin out the original point clouds while retaining characteristic features, we resort to extracting 3D keypoints. Such clouds of keypoints, which can be viewed as a sparse but nevertheless discriminative representation of the original scans, are then used as input to a very efficient matching method originally developed in computer graphics, called 4-Points Congruent Sets (4PCS) algorithm. We adapt the 4PCS matching approach to better suit the characteristics of laser scans. The resulting Keypoint-based 4-Points Congruent Sets (K-4PCS) method is extensively evaluated on challenging indoor and outdoor scans. Beyond the evaluation on real terrestrial laser scans, we also perform experiments with simulated indoor scenes, paying particular attention to the sensitivity of the approach with respect to highly symmetric scenes.

  4. Function analysis of working integrated circuit with scanning laser microscope. Laser kenbikyo ni yoru IC no dosa kansatsu

    Energy Technology Data Exchange (ETDEWEB)

    Ode, T. (Lasertec Corp., Kanagawa (Japan))

    1992-10-20

    By scanning a laser light, the reaction of a specimen against the light is detected in some means. The optical effect can be visualized by displaying that on the CRT or the like in synchronism with the scanning. Among these, an image formed and visualized by internal photoelectric effect by light is called OBIC image, and chiefly used for evaluating and analyzing semiconductor devices. Observing this OBIC image by a high speed scanning laser microscope has been spotlighted these days as an effective means for observing the state of p-n junction of an IC in operation. This paper descries the principle, the observing method, the detecting circuit, etc. of the semiconductor observing method using a laser microscope. Further, actual examples of detecting defects of an IC by means of OBIC image are shown. As for the problem, since leak parts are displayed as negative contrast in the OBIC image to affect finding work of leak part, the necessity of improvement is pointed out. 39 refs., 11 figs.

  5. Comparison of 3d Reconstruction Services and Terrestrial Laser Scanning for Cultural Heritage Documentation

    Science.gov (United States)

    Rasztovits, S.; Dorninger, P.

    2013-07-01

    Terrestrial Laser Scanning (TLS) is an established method to reconstruct the geometrical surface of given objects. Current systems allow for fast and efficient determination of 3D models with high accuracy and richness in detail. Alternatively, 3D reconstruction services are using images to reconstruct the surface of an object. While the instrumental expenses for laser scanning systems are high, upcoming free software services as well as open source software packages enable the generation of 3D models using digital consumer cameras. In addition, processing TLS data still requires an experienced user while recent web-services operate completely automatically. An indisputable advantage of image based 3D modeling is its implicit capability for model texturing. However, the achievable accuracy and resolution of the 3D models is lower than those of laser scanning data. Within this contribution, we investigate the results of automated web-services for image based 3D model generation with respect to a TLS reference model. For this, a copper sculpture was acquired using a laser scanner and using image series of different digital cameras. Two different webservices, namely Arc3D and AutoDesk 123D Catch were used to process the image data. The geometric accuracy was compared for the entire model and for some highly structured details. The results are presented and interpreted based on difference models. Finally, an economical comparison of the generation of the models is given considering the interactive and processing time costs.

  6. Parallel detection experiment of fluorescence confocal microscopy using DMD.

    Science.gov (United States)

    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.

  7. In vivo confocal microscopy in dermatology: from research to clinical application

    Science.gov (United States)

    Ulrich, Martina; Lange-Asschenfeldt, Susanne

    2013-06-01

    Confocal laser scanning microscopy (CLSM) represents an emerging technique for the noninvasive histomorphological analysis of skin in vivo and has shown its applicability for dermatological research as well as its value as an adjunct tool in the clinical management of skin cancer patients. Herein, we aim to give an overview on the current clinical indications for CLSM in dermatology and also highlight the diverse applications of CLSM in dermatological research.

  8. Chondrocytes provide a model for in-situ confocal microscopy and 3D reconstructions

    Science.gov (United States)

    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. 3D laser scanning and modelling of the Dhow heritage for the Qatar National Museum

    Science.gov (United States)

    Wetherelt, A.; Cooper, J. P.; Zazzaro, C.

    2014-08-01

    Curating boats can be difficult. They are complex structures, often demanding to conserve whether in or out of the water; they are usually large, difficult to move on land, and demanding of gallery space. Communicating life on board to a visiting public in the terra firma context of a museum can be difficult. Boats in their native environment are inherently dynamic artifacts. In a museum they can be static and divorced from the maritime context that might inspire engagement. New technologies offer new approaches to these problems. 3D laser scanning and digital modeling offers museums a multifaceted means of recording, monitoring, studying and communicating watercraft in their care. In this paper we describe the application of 3D laser scanning and subsequent digital modeling. Laser scans were further developed using computer-generated imagery (CGI) modeling techniques to produce photorealistic 3D digital models for development into interactive, media-based museum displays. The scans were also used to generate 2D naval lines and orthographic drawings as a lasting curatorial record of the dhows held by the National Museum of Qatar.

  10. Inspection of float glass using a novel retroreflective laser scanning system

    Science.gov (United States)

    Holmes, Jonathan D.

    1997-07-01

    Since 1988, Image Automation has marketed a float glass inspection system using a novel retro-reflective laser scanning system. The (patented) instrument scans a laser beam by use of a polygon through the glass onto a retro-reflective screen, and collects the retro-reflected light off the polygon, such that a stationary image of the moving spot on the screen is produced. The spot image is then analyzed for optical effects introduced by defects within the glass, which typically distort and attenuate the scanned laser beam, by use of suitable detectors. The inspection system processing provides output of defect size, shape and severity, to the factory network for use in rejection or sorting of glass plates to the end customer. This paper briefly describes the principles of operation, the system architecture, and limitations to sensitivity and measurement repeatability. New instruments based on the retro-reflective scanning method have recently been developed. The principles and implementation are described. They include: (1) Simultaneous detection of defects within the glass and defects in a mirror coating on the glass surface using polarized light. (2) A novel distortion detector for very dark glass. (3) Measurement of optical quality (flatness/refractive homogeneity) of the glass using a position sensitive detector.

  11. Wafer-level vacuum packaged resonant micro-scanning mirrors for compact laser projection displays

    Science.gov (United States)

    Hofmann, Ulrich; Oldsen, Marten; Quenzer, Hans-Joachim; Janes, Joachim; Heller, Martin; Weiss, Manfred; Fakas, Georgios; Ratzmann, Lars; Marchetti, Eleonora; D'Ascoli, Francesco; Melani, Massimiliano; Bacciarelli, Luca; Volpi, Emilio; Battini, Francesco; Mostardini, Luca; Sechi, Francesco; De Marinis, Marco; Wagner, Bernd

    2008-02-01

    Scanning laser projection using resonant actuated MEMS scanning mirrors is expected to overcome the current limitation of small display size of mobile devices like cell phones, digital cameras and PDAs. Recent progress in the development of compact modulated RGB laser sources enables to set up very small laser projection systems that become attractive not only for consumer products but also for automotive applications like head-up and dash-board displays. Within the last years continuous progress was made in increasing MEMS scanner performance. However, only little is reported on how mass-produceability of these devices and stable functionality even under harsh environmental conditions can be guaranteed. Automotive application requires stable MEMS scanner operation over a wide temperature range from -40° to +85°Celsius. Therefore, hermetic packaging of electrostatically actuated MEMS scanning mirrors becomes essential to protect the sensitive device against particle contamination and condensing moisture. This paper reports on design, fabrication and test of a resonant actuated two-dimensional micro scanning mirror that is hermetically sealed on wafer level. With resonant frequencies of 30kHz and 1kHz, an achievable Theta-D-product of 13mm.deg and low dynamic deformation <20nm RMS it targets Lissajous projection with SVGA-resolution. Inevitable reflexes at the vacuum package surface can be seperated from the projection field by permanent inclination of the micromirror.

  12. Nano-pulsed laser irradiation scanning system for phase-change materials

    International Nuclear Information System (INIS)

    Kim, Sookyung; Li Xuezhe; Lee, Sangbin; Kim, Kyung-Ho; Lee, Seung-Yop

    2008-01-01

    Recently, the demand of a laser irradiation tester is increasing for phase change random access memory (PRAM) as well as conventional optical storage media. In this study, a nano-pulsed laser irradiation system is developed to characterize the optical property and writing performance of phase-change materials, based on a commercially available digital versatile disk (DVD) optical pick-up. The precisely controlled focusing and scanning on the material's surface are implemented using the auto-focusing mechanism and a voice coil motor (VCM) of the commercial DVD pick-up. The laser irradiation system provides various writing and reading functions such as adjustable laser power, pulse duration, recording pattern (spot, line and area), and writing/reading repetition, phase transition, and in situ reflectivity measurement before/after irradiation. Measurements of power time effect (PTE) diagram and reflectivity map of Ge 2 Sb 2 Te 5 samples show that the proposed laser irradiation system provides the powerful scanning tool to quantify the optical characteristics of phase-change materials

  13. Laser scanning endoscope via an imaging fiber bundle for fluorescence imaging

    Science.gov (United States)

    Yeboah, Lorenz D.; Nestler, Dirk; Steiner, Rudolf W.

    1994-12-01

    Based on a laser scanning endoscope via an imaging fiber bundle, a new approach for a tumor diagnostic system has been developed to assist physicians in the diagnosis before the actual PDT is carried out. Laser induced, spatially resolved fluorescence images of diseased tissue can be compared with images received by video endoscopy using a white light source. The set- up is required to produce a better contrast between infected and healthy tissue and might serve as a constructive diagnostic help for surgeons. The fundamental idea is to scan a low-power laser beam on an imaging fiber bundle and to achieve a spatially resolved projection on the tissue surface. A sufficiently high laser intensity from the diode laser is concentrated on each single spot of the tissue exciting fluorescence when a dye has previously been accumulated. Subsequently, video image of the tissue is recorded and stored. With an image processing unit, video and fluorescence images are overlaid producing a picture of the fluorescence intensity in the environment of the observed tissue.

  14. Generation of 3D Virtual Geographic Environment Based on Laser Scanning Technique

    Institute of Scientific and Technical Information of China (English)

    DU Jie; CHEN Xiaoyong; FumioYamazaki

    2003-01-01

    This paper demonstrates an experiment on the generation of 3D virtual geographic environment on the basis of experimental flight laser scanning data by a set of algorithms and methods that were developed to automatically interpret range images for extracting geo-spatial features and then to reconstruct geo-objects. The algorithms and methods for the interpretation and modeling of laser scanner data include triangulated-irregular-network (TIN)-based range image interpolation ; mathematical-morphology(MM)-based range image filtering,feature extraction and range image segmentation, feature generalization and optimization, 3D objects reconstruction and modeling; computergraphics (CG)-based visualization and animation of geographic virtual reality environment.

  15. Brief communication "Application of mobile laser scanning in snow cover profiling"

    Directory of Open Access Journals (Sweden)

    S. Kaasalainen

    2011-03-01

    Full Text Available We present a snowmobile-based mobile mapping system and its first application to snow cover roughness and change detection measurement. The ROAMER mobile mapping system, constructed at the Finnish Geodetic Institute, consists of the positioning and navigating systems, a terrestrial laser scanner, and the carrying platform (a snowmobile sledge in this application. We demonstrate the applicability of the instrument to snow cover roughness profiling and change detection by presenting preliminary results from a mobile laser scanning (MLS campaign. The results show the potential of MLS for fast and efficient snow profiling from large areas in a millimetre scale.

  16. The Investigation of Accuracy of 3 Dimensional Models Generated From Point Clouds with Terrestrial Laser Scanning

    Science.gov (United States)

    Gumus, Kutalmis; Erkaya, Halil

    2013-04-01

    In Terrestrial laser scanning (TLS) applications, it is necessary to take into consideration the conditions that affect the scanning process, especially the general characteristics of the laser scanner, geometric properties of the scanned object (shape, size, etc.), and its spatial location in the environment. Three dimensional models obtained with TLS, allow determining the geometric features and relevant magnitudes of the scanned object in an indirect way. In order to compare the spatial location and geometric accuracy of the 3-dimensional model created by Terrestrial laser scanning, it is necessary to use measurement tools that give more precise results than TLS. Geometric comparisons are performed by analyzing the differences between the distances, the angles between surfaces and the measured values taken from cross-sections between the data from the 3-dimensional model created with TLS and the values measured by other measurement devices The performance of the scanners, the size and shape of the scanned objects are tested using reference objects the sizes of which are determined with high precision. In this study, the important points to consider when choosing reference objects were highlighted. The steps up to processing the point clouds collected by scanning, regularizing these points and modeling in 3 dimensions was presented visually. In order to test the geometric correctness of the models obtained by Terrestrial laser scanners, sample objects with simple geometric shapes such as cubes, rectangular prisms and cylinders that are made of concrete were used as reference models. Three dimensional models were generated by scanning these reference models with Trimble Mensi GS 100. The dimension of the 3D model that is created from point clouds was compared with the precisely measured dimensions of the reference objects. For this purpose, horizontal and vertical cross-sections were taken from the reference objects and generated 3D models and the proximity of

  17. Probing the cellular damage in bacteria induced by GaN nanoparticles using confocal laser Raman spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Sahoo, Prasana, E-mail: prasanasahoo@gmail.com [Indira Gandhi Center for Atomic Research, Surface and Nanoscience Division (India); Murthy, P. Sriyutha [Bhabha Atomic Research Centre, Biofouling and Biofilm Processes Section, Water and Steam Chemistry Division (India); Dhara, S., E-mail: dhara@igcar.gov.in [Indira Gandhi Center for Atomic Research, Surface and Nanoscience Division (India); Venugopalan, V. P. [Bhabha Atomic Research Centre, Biofouling and Biofilm Processes Section, Water and Steam Chemistry Division (India); Das, A.; Tyagi, A. K. [Indira Gandhi Center for Atomic Research, Surface and Nanoscience Division (India)

    2013-08-15

    Understanding the mechanism of nanoparticle (NP) induced toxicity in microbes is of potential importance to a variety of disciplines including disease diagnostics, biomedical implants, and environmental analysis. In this context, toxicity to bacterial cells and inhibition of biofilm formation by GaN NPs and their functional derivatives have been investigated against gram positive and gram negative bacterial species down to single cellular level. High levels of inhibition of biofilm formation (>80 %) was observed on treatments with GaN NPs at sub-micro molar concentrations. These results were substantiated with morphological features investigated with field emission scanning electron microscope, and the observed changes in vibrational modes of microbial cells using Raman spectroscopy. Raman spectra provided molecular interpretation of cell damage by registering signatures of molecular vibrations of individual living microbial cells and mapping the interplay of proteins at the cell membrane. As compared to the untreated cells, Raman spectra of NP-treated cells showed an increase in the intensities of characteristic protein bands, which confirmed membrane damage and subsequent release of cellular contents outside the cells. Raman spectral mapping at single cellular level can facilitate understanding of the mechanistic aspect of toxicity of GaN NPs. The effect may be correlated to passive diffusion causing mechanical damage to the membrane or ingress of Ga{sup 3+} (ionic radius {approx}0.076 nm) which can potentially interfere with bacterial metabolism, as it resembles Fe{sup 2+} (ionic radius {approx}0.077 nm), which is essential for energy metabolism.

  18. Probing the cellular damage in bacteria induced by GaN nanoparticles using confocal laser Raman spectroscopy

    International Nuclear Information System (INIS)

    Sahoo, Prasana; Murthy, P. Sriyutha; Dhara, S.; Venugopalan, V. P.; Das, A.; Tyagi, A. K.

    2013-01-01

    Understanding the mechanism of nanoparticle (NP) induced toxicity in microbes is of potential importance to a variety of disciplines including disease diagnostics, biomedical implants, and environmental analysis. In this context, toxicity to bacterial cells and inhibition of biofilm formation by GaN NPs and their functional derivatives have been investigated against gram positive and gram negative bacterial species down to single cellular level. High levels of inhibition of biofilm formation (>80 %) was observed on treatments with GaN NPs at sub-micro molar concentrations. These results were substantiated with morphological features investigated with field emission scanning electron microscope, and the observed changes in vibrational modes of microbial cells using Raman spectroscopy. Raman spectra provided molecular interpretation of cell damage by registering signatures of molecular vibrations of individual living microbial cells and mapping the interplay of proteins at the cell membrane. As compared to the untreated cells, Raman spectra of NP-treated cells showed an increase in the intensities of characteristic protein bands, which confirmed membrane damage and subsequent release of cellular contents outside the cells. Raman spectral mapping at single cellular level can facilitate understanding of the mechanistic aspect of toxicity of GaN NPs. The effect may be correlated to passive diffusion causing mechanical damage to the membrane or ingress of Ga 3+ (ionic radius ∼0.076 nm) which can potentially interfere with bacterial metabolism, as it resembles Fe 2+ (ionic radius ∼0.077 nm), which is essential for energy metabolism

  19. A novel method for enhancing the lateral resolution and image SNR in confocal microscopy

    Science.gov (United States)

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

    2017-12-01

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

  20. Preliminary results on the anatomy of the larval musculature of Balanus improvisus (Darwin, 1854) (Crustacea: Cirripedia: Thecostraca) using phalloidin staining in combination with confocal laserscanning microscopy

    DEFF Research Database (Denmark)

    Semmler, Henrike; Høeg, Jens Thorvald; Scholtz, Gerhard

    2006-01-01

    The anatomy of the larval muscular systems in Balanus improvisus (Darwin, 1854) was investigated by using phalloidin staining to visualize filamentous F-actin in combination with confocal laser scanning microscopy (CLSM). The larval musculature contains an anterior muscle complex associated...

  1. Multi-objective optimization of cellular scanning strategy in selective laser melting

    DEFF Research Database (Denmark)

    Ahrari, Ali; Deb, Kalyanmoy; Mohanty, Sankhya

    2017-01-01

    The scanning strategy for selective laser melting - an additive manufacturing process - determines the temperature fields during the manufacturing process, which in turn affects residual stresses and distortions, two of the main sources of process-induced defects. The goal of this study is to dev......The scanning strategy for selective laser melting - an additive manufacturing process - determines the temperature fields during the manufacturing process, which in turn affects residual stresses and distortions, two of the main sources of process-induced defects. The goal of this study......, the problem is a combination of combinatorial and choice optimization, which makes the problem difficult to solve. On a process simulation domain consisting of 32 cells, our multi-objective evolutionary method is able to find a set of trade-off solutions for the defined conflicting objectives, which cannot...

  2. Development of Large Concrete Object Geometrical Model Based on Terrestrial Laser Scanning

    Directory of Open Access Journals (Sweden)

    Zaczek-Peplinska Janina

    2015-02-01

    Full Text Available The paper presents control periodic measurements of movements and survey of concrete dam on Dunajec River in Rożnów, Poland. Topographical survey was conducted using laser scanning technique. The goal of survey was data collection and creation of a geometrical model. Acquired cross- and horizontal sections were utilised to create a numerical model of object behaviour at various load depending of changing level of water in reservoir. Modelling was accomplished using finite elements technique. During the project an assessment was conducted to terrestrial laser scanning techniques for such type of research of large hydrotechnical objects such as gravitational water dams. Developed model can be used to define deformations and displacement prognosis.

  3. Microstructures and Microhardness Properties of CMSX-4® Additively Fabricated Through Scanning Laser Epitaxy (SLE)

    Science.gov (United States)

    Basak, Amrita; Holenarasipura Raghu, Shashank; Das, Suman

    2017-12-01

    Epitaxial CMSX-4® deposition is achieved on CMSX-4® substrates through the scanning laser epitaxy (SLE) process. A thorough analysis is performed using various advanced material characterization techniques, namely high-resolution optical microscopy, scanning electron microscopy, energy-dispersive x-ray spectroscopy, x-ray diffraction, and Vickers microhardness measurements, to characterize and compare the quality of the SLE-fabricated CMSX-4® deposits to the CMSX-4® substrates. The results show that the CMSX-4® deposits have smaller primary dendritic arm spacing, finer γ/ γ' size, weaker elemental segregation, and higher microhardness compared to the investment cast CMSX-4® substrates. The results presented here demonstrate that CMSX-4® is an attractive material for laser-based AM processing and, therefore, can be used in the fabrication of gas turbine hot-section components through AM processing.

  4. As- built inventory of the office building with the use of terrestrial laser scanning

    Science.gov (United States)

    Przyborski, Marek; Tysiąc, Paweł

    2018-01-01

    Terrestrial Laser Scanning (TLS) is an efficient tool for building inventories. Based on the red- laser beam technology it is possible to provide the high accuracy data with complete spatial information about a scanned object. In this article, authors present the solution of use a TLS in as-built inventory of the office building. Based on the provided data, it is possible to evaluate the correctness of built details of a building and provide information for further construction works, for example an area needed for Styrofoam installation. The biggest problem in this research is that an error which equals over 1cm could generate costs, which could be a problem to cover by a constructor. Based on a complicated place of the construction works (centre of a city) it was a challenge to maintain the accuracy.

  5. As- built inventory of the office building with the use of terrestrial laser scanning

    Directory of Open Access Journals (Sweden)

    Przyborski Marek

    2018-01-01

    Full Text Available Terrestrial Laser Scanning (TLS is an efficient tool for building inventories. Based on the red- laser beam technology it is possible to provide the high accuracy data with complete spatial information about a scanned object. In this article, authors present the solution of use a TLS in as-built inventory of the office building. Based on the provided data, it is possible to evaluate the correctness of built details of a building and provide information for further construction works, for example an area needed for Styrofoam installation. The biggest problem in this research is that an error which equals over 1cm could generate costs, which could be a problem to cover by a constructor. Based on a complicated place of the construction works (centre of a city it was a challenge to maintain the accuracy.

  6. Damage Detection on Thin-walled Structures Utilizing Laser Scanning and Standing Waves

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Se Hyeok; Jeon, Jun Young; Kim, Du Hwan; Park, Gyuhae [Chonnam Nat’l Univ., Gwangju (Korea, Republic of); Kang, To; Han, Soon Woo [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2017-05-15

    This paper describes wavenumber filtering for damage detection using single-frequency standing wave excitation and laser scanning sensing. An embedded piezoelectric sensor generates ultrasonic standing waves, and the responses are measured using a laser Doppler vibrometer and mirror tilting device. After scanning, newly developed damage detection techniques based on wavenumber filtering are applied to the full standing wave field. To demonstrate the performance of the proposed techniques, several experiments were performed on composite plates with delamination and aluminum plates with corrosion damage. The results demonstrated that the developed techniques could be applied to various structures to localize the damage, with the potential to improve the damage detection capability at a high interrogation speed.

  7. In vivo visualization of microneedle conduits in human skin using laser scanning microscopy

    International Nuclear Information System (INIS)

    Bal, S; Kruithof, A C; Bouwstra, J; Liebl, H; Tomerius, M; Lademann, J; Meinke, M

    2010-01-01

    Solid microneedles enhance the penetration of drugs into the viable skin but little is known about the geometry of the conduits in vivo. Therefore, laser scanning microscopy was used to visualize the conduits of a microneedle system with needles at a length of 300 μm in 6 healthy subjects over a period of time. The model drug, a fluorescent dye was applied before and after piercing. Laser scanning microscopy was evaluated as being an excellent method to monitor the geometry and closure of the conduits over time. The used microneedle system was evaluated as suitable to enhance the transport of model drugs into the viable epidermis without bleeding and a short closure time of the conduits at the skin surface

  8. In vivo visualization of microneedle conduits in human skin using laser scanning microscopy

    Science.gov (United States)

    Bal, S.; Kruithof, A. C.; Liebl, H.; Tomerius, M.; Bouwstra, J.; Lademann, J.; Meinke, M.

    2010-03-01

    Solid microneedles enhance the penetration of drugs into the viable skin but little is known about the geometry of the conduits in vivo. Therefore, laser scanning microscopy was used to visualize the conduits of a microneedle system with needles at a length of 300 μm in 6 healthy subjects over a period of time. The model drug, a fluorescent dye was applied befor