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...

Update on Development of the Potassium-Argon Laser Experiment (KArLE) Instrument for In Situ Geochronology

Science.gov (United States)

Cohen, Barbara A.; Li, Z.-H.; Miller, J. S.; Brinckerhoff, W. B.; Clegg, S. M.; Mahaffy, P. R.; Swindle, T. D.; Wiens, R. C.

2013-01-01

Absolute dating of planetary samples is an essential tool to establish the chronology of geological events, including crystallization history, magmatic evolution, and alteration. We are addressing this challenge by developing the Potassium (K) -- Argon Laser Experiment (KArLE), building on previous work to develop a K-Ar in situ instrument. KArLE ablates a rock sample, determines the K in the plasma state using laser-induced breakdown spectroscopy (LIBS), measures the liberated Ar using quadrupole mass spectrometry (QMS), and relates the two by the volume of the ablated pit using laser confocal microscopy (LCM). Our goal is for the KArLE instrument to be capable of determining the age of several kinds of planetary samples to address a wide range of geochronolgy problems in planetary science.

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.

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...

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.

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.

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...

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.

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.

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.

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

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

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.

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...

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

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.

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.

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.

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.

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.

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.

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

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

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.

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.)

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....

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.

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.

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.

Laser light scattering instrument advanced technology development

Science.gov (United States)

Wallace, J. F.

1993-01-01

The objective of this advanced technology development (ATD) project has been to provide sturdy, miniaturized laser light scattering (LLS) instrumentation for use in microgravity experiments. To do this, we assessed user requirements, explored the capabilities of existing and prospective laser light scattering hardware, and both coordinated and participated in the hardware and software advances needed for a flight hardware instrument. We have successfully breadboarded and evaluated an engineering version of a single-angle glove-box instrument which uses solid state detectors and lasers, along with fiber optics, for beam delivery and detection. Additionally, we have provided the specifications and written verification procedures necessary for procuring a miniature multi-angle LLS instrument which will be used by the flight hardware project which resulted from this work and from this project's interaction with the laser light scattering community.

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)

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).

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.

Conduction cooled compact laser for chemcam instrument

Science.gov (United States)

Faure, B.; Saccoccio, M.; Maurice, S.; Durand, E.; Derycke, C.

2017-11-01

A new conduction cooled compact laser for Laser Induced Breakdown Spectroscopy (LIBS) on Mars is presented. The laser provides pulses with energy higher than 30mJ at 1μm of wavelength with a good spatial quality. Three development prototypes of this laser have been built and functional and environmental tests have been done. Then, the Qualification and Flight models have been developed and delivered. A spare model is now developed. This laser will be mounted on the ChemCam Instrument of the NASA mission MSL 2009. ChemCam Instrument is developed in collaboration between France (CESR and CNES) and USA (LANL). The goal of this Instrument is to study the chemical composition of Martian rocks. A laser source (subject of this presentation) emits a pulse which is focused by a telescope. It creates a luminous plasma on the rock; the light of this plasma is then analysed by three spectrometers to obtain information on the composition of the rock. The laser source is developed by the French company Thales Laser, with a technical support from CNES and CESR. This development is funded by CNES. The laser is compact, designed to work in burst mode. It doesn't require any active cooling.

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.

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

Increasing Laser Stability with Improved Electronic Instruments

Science.gov (United States)

Troxel, Daylin; Bennett, Aaron; Erickson, Christopher J.; Jones, Tyler; Durfee, Dallin S.

2010-03-01

We present several electronic instruments developed to implement an ultra-stable laser lock. These instruments include a high speed, low noise homodyne photo-detector; an ultrahigh stability, low noise current driver with high modulation bandwidth and digital control; a high-speed, low noise PID controller; a low-noise piezo driver; and a laser diode temperature controller. We will present the theory of operation for these instruments, design and construction techniques, and essential characteristics for each device.

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.

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...

Advancement of High Power Quasi-CW Laser Diode Arrays For Space-based Laser Instruments

Science.gov (United States)

Amzajerdian, Farzin; Meadows, Byron L.; Baker, nathaniel R.; Baggott, Renee S.; Singh, Upendra N.; Kavaya, Michael J.

2004-01-01

Space-based laser and lidar instruments play an important role in NASA s plans for meeting its objectives in both Earth Science and Space Exploration areas. Almost all the lidar instrument concepts being considered by NASA scientist utilize moderate to high power diode-pumped solid state lasers as their transmitter source. Perhaps the most critical component of any solid state laser system is its pump laser diode array which essentially dictates instrument efficiency, reliability and lifetime. For this reason, premature failures and rapid degradation of high power laser diode arrays that have been experienced by laser system designers are of major concern to NASA. This work addresses these reliability and lifetime issues by attempting to eliminate the causes of failures and developing methods for screening laser diode arrays and qualifying them for operation in space.

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.

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.

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.

Angle measurement with laser feedback instrument.

Science.gov (United States)

Chen, Wenxue; Zhang, Shulian; Long, Xingwu

2013-04-08

An instrument for angle measurement based on laser feedback has been designed. The measurement technique is based on the principle that when a wave plate placed into a feedback cavity rotates, its phase retardation varies. Phase retardation is a function of the rotating angle of the wave plate. Hence, the angle can be converted to phase retardation. The phase retardation is measured at certain characteristic points identified in the laser outputting curve that are then modulated by laser feedback. The angle of a rotating object can be measured if it is connected to the wave plate. The main advantages of this instrument are: high resolution, compact, flexible, low cost, effective power, and fast response.

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

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.

Design of instrumentation and software for precise laser machining

Science.gov (United States)

Wyszyński, D.; Grabowski, Marcin; Lipiec, Piotr

2017-10-01

The paper concerns the design of instrumentation and software for precise laser machining. Application of advanced laser beam manipulation instrumentation enables noticeable improvement of cut quality and material loss. This factors have significant impact on process efficiency and cutting edge quality by means of machined part size and shape accuracy, wall taper, material loss reduction (e.g. diamond) and time effectiveness. The goal can be reached by integration of laser drive, observation and optical measurement system, beam manipulation system and five axis mechanical instrumentation with use of advanced tailored software enabling full laser cutting process control and monitoring.

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...

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

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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.

21 CFR 886.1360 - Visual field laser instrument.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Visual field laser instrument. 886.1360 Section 886.1360 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Diagnostic Devices § 886.1360 Visual field laser instrument...

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.)

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.

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...

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...

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

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.

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.

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

Science.gov (United States)

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

1999-11-01

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

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)

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.

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...

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.

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.

Signal and noise modeling in confocal laser scanning fluorescence microscopy.

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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.

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...

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.

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.

Confocal endomicroscopy: Is it time to move on?

Science.gov (United States)

Robles-Medranda, Carlos

2016-01-10

Confocal laser endomicroscopy permits in-vivo microscopy evaluation during endoscopy procedures. It can be used in all the parts of the gastrointestinal tract and includes: Esophagus, stomach, small bowel, colon, biliary tract through and endoscopic retrograde cholangiopancreatography and pancreas through needles during endoscopic ultrasound procedures. Many researches demonstrated a high correlation of results between confocal laser endomicroscopy and histopathology in the diagnosis of gastrointestinal lesions; with accuracy in about 86% to 96%. Moreover, in spite that histopathology remains the gold-standard technique for final diagnosis of any diseases; a considerable number of misdiagnosis rate could be present due to many factors such as interpretation mistakes, biopsy site inaccuracy, or number of biopsies. Theoretically; with the diagnostic accuracy rates of confocal laser endomicroscopy could help in a daily practice to improve diagnosis and treatment management of the patients. However, it is still not routinely used in the clinical practice due to many factors such as cost of the procedure, lack of codification and reimbursement in some countries, absence of standard of care indications, availability, physician image-interpretation training, medico-legal problems, and the role of the pathologist. These limitations are relative, and solutions could be found based on new researches focused to solve these barriers.

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.

Confocal Raman microscopy for in depth analysis in the field of cultural heritage

Science.gov (United States)

Lorenzetti, G.; Striova, J.; Zoppi, A.; Castellucci, E. M.

2011-05-01

In the field of cultural heritage, the main concern when a sample is analyzed is its safeguard, and this means that non-destructive techniques are required. In this work, we show how confocal Raman microscopy (CRM) may be successfully applied in the study of works of art as a valuable alternative to other well established techniques. CRM with a metallurgical objective was tested for the in depth study of thin samples that are of interest in the field of cultural heritage. The sensitivity of the instrumentation was first evaluated by analyzing single layers of pure polyethylene terephthalate (PET) films having a thickness of 12, 25, and 50 μm, respectively, and a multilayer sample of polypropylene (PP) and polyethylene (PE). Subsequently, the technique was applied to the analysis of historical dyed cotton yarns in order to check whether it was possible to achieve a better discrimination of the fibres' signals for an easier identification. A substantial improvement of the signal to noise ratio was found in the confocal arrangement with respect to the non-confocal one, suggesting the use of this technique for this kind of analysis in the field of cultural heritage. Furthermore, Raman spectroscopy in confocal configuration was exploited in the evaluation of cleaning performed on the mural painting specimens, treated with acrylic resin (Paraloid B72). Confocal Raman experiments were performed before and after laser cleaning (at different conditions) in order to monitor the presence and to approximate the polymer thickness: the method proved to be a valid comparative tool in assessment of cleaning efficiencies.

Confocal Raman Microscopy

CERN Document Server

Dieing, Thomas; Toporski, Jan

2011-01-01

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

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.

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)

Mineralogy and Astrobiology Detection Using Laser Remote Sensing Instrument

Science.gov (United States)

Abedin, M. Nurul; Bradley, Arthur T.; Sharma, Shiv K.; Misra, Anupam K.; Lucey, Paul G.; Mckay, Chistopher P.; Ismail, Syed; Sandford, Stephen P.

2015-01-01

A multispectral instrument based on Raman, laser-induced fluorescence (LIF), laser-induced breakdown spectroscopy (LIBS), and a lidar system provides high-fidelity scientific investigations, scientific input, and science operation constraints in the context of planetary field campaigns with the Jupiter Europa Robotic Lander and Mars Sample Return mission opportunities. This instrument conducts scientific investigations analogous to investigations anticipated for missions to Mars and Jupiter's icy moons. This combined multispectral instrument is capable of performing Raman and fluorescence spectroscopy out to a >100 m target distance from the rover system and provides single-wavelength atmospheric profiling over long ranges (>20 km). In this article, we will reveal integrated remote Raman, LIF, and lidar technologies for use in robotic and lander-based planetary remote sensing applications. Discussions are focused on recently developed Raman, LIF, and lidar systems in addition to emphasizing surface water ice, surface and subsurface minerals, organics, biogenic, biomarker identification, atmospheric aerosols and clouds distributions, i.e., near-field atmospheric thin layers detection for next robotic-lander based instruments to measure all the above-mentioned parameters. OCIS codes: (120.0280) Remote sensing and sensors; (130.0250) Optoelectronics; (280.3640) Lidar; (300.2530) Fluorescence, laser-induced; (300.6450) Spectroscopy, Raman; (300.6365) Spectroscopy, laser induced breakdown

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.

  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...

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)

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.

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

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.)

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.

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.

Comparison between carbondioxide laser and cold instruments in treatment of vocal nodule

International Nuclear Information System (INIS)

Kundi, N.A.; Qayyum, A.; Ahmed, B.; Raza, M.

2013-01-01

Background: Vocal cord nodules are one of the most frequent disorders in both children and adults who use their voice excessively. Main symptom with which patient presents is hoarseness of voice. The treatment in early stages is voice therapy. Various methods are used for its treatment e.g. surgical removal with cold instruments and carbon dioxide laser ablation. Response to the treatment is measured by improvement in voice quality. Objective: To compare the results of Carbon dioxide laser and cold instruments in the treatment of vocal nodule. Study Design: Quasi-experimental study. Place and Duration of Study: This study was conducted at Otolaryngology Department Combined Military Hospital Rawalpindi. Patients and Methods: In this study 50 patients undergoing treatment of vocal cord nodule were included, 25 patients were treated by surgical removal with cold instruments and 25 patients were treated with carbon dioxide laser ablation. The main comparative outcomes were measured by patients' perception of voice quality (worse, same, improved) one week post operatively. Results: Improvement in voice quality with carbon dioxide laser was found to be clinically superior. Voice quality was significantly improved as compared to cold surgical instruments. Conclusion: Carbon dioxide laser causes early improvement in quality of voice as compared to cold instruments in the treatment of vocal nodules. (author)

Living Matter Observations with a Novel Hyperspectral Supercontinuum Confocal Microscope for VIS to Near-IR Reflectance Spectroscopy

Directory of Open Access Journals (Sweden)

Francesca R. Bertani

2013-10-01

Full Text Available A broad range hyper-spectroscopic microscope fed by a supercontinuum laser source and equipped with an almost achromatic optical layout is illustrated with detailed explanations of the design, implementation and data. The real novelty of this instrument, a confocal spectroscopic microscope capable of recording high resolution reflectance data in the VIS-IR spectral range from about 500 nm to 2.5 μm wavelengths, is the possibility of acquiring spectral data at every physical point as defined by lateral coordinates, X and Y, as well as at a depth coordinate, Z, as obtained by the confocal optical sectioning advantage. With this apparatus we collect each single scanning point as a whole spectrum by combining two linear spectral detector arrays, one CCD for the visible range, and one InGaAs infrared array, simultaneously available at the sensor output channel of the home made instrument. This microscope has been developed for biomedical analysis of human skin and other similar applications. Results are shown illustrating the technical performances of the instrument and the capability in extracting information about the composition and the structure of different parts or compartments in biological samples as well as in solid statematter. A complete spectroscopic fingerprinting of samples at microscopic level is shown possible by using statistical analysis on raw data or analytical reflectance models based on Abelés matrix transfer methods.

Conversion efficiency of implanted ions by confocal micro-luminescence mapping

International Nuclear Information System (INIS)

Deshko, Y.; Huang, Mengbing; Gorokhovsky, A.A.

2013-01-01

We report on the further development of the statistical approach to determine the conversion efficiency of implanted ions into emitting centers and present the measurement method based on the confocal micro-luminescence mapping. It involves the micro-luminescence mapping with a narrow-open confocal aperture, followed by the statistical analysis of the photoluminescence signal from an ensemble of emitting centers. The confocal mapping method has two important advantages compared to the recently discussed aperture-free method (J. Lumin. 131 (2011) 489): it is less sensitive to errors in the laser spot size and has a well defined useful area. The confocal mapping has been applied to the Xe center in diamond. The conversion efficiency has been found to be about 0.28, which is in good agreement with the results of the aperture-free method. - Highlights: ► Conversion efficiency of implanted ions into emitting centers – statistical approach. ► Micro-luminescence mapping with open and narrow confocal aperture – comparison. ► Advantages of the confocal micro-luminescence mapping. ► Confocal micro-luminescence mapping has been applied to the Xe center in diamond. ► The conversion efficiency has been found to be about 0.28.

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.

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.

The Mercury Laser Altimeter Instrument for the MESSENGER Mission

Science.gov (United States)

Cavanaugh, John F.; Smith, James C.; Sun, Xiaoli; Bartels, Arlin E.; Ramos-Izquierdo, Luis; Krebs, Danny J.; Novo-Gradac, Anne marie; McGarry, Jan F.; Trunzo, Raymond; Britt, Jamie L.

2006-01-01

The Mercury Laser Altimeter (MLA) is one of the payload science instruments on the MErcury Surface, Space ENvironment, GEochemistry, and Ranging (MESSENGER) mission, which launched on 3 August 2004. The altimeter will measure the round trip time-of-flight of transmitted laser pulses reflected from the surface of the planet that, in combination with the spacecraft orbit position and pointing data, gives a high-precision measurement of surface topography referenced to Mercury's center of mass. The altimeter measurements will be used to determine the planet's forced librations by tracking the motion of large-scale topographic features as a function of time. MLA's laser pulse energy monitor and the echo pulse energy estimate will provide an active measurement of the surface reflectivity at 1064 nm. This paper describes the instrument design, prelaunch testing, calibration, and results of post-launch testing.

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...

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.

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.

Multimode laser beam analyzer instrument using electrically programmable optics.

Science.gov (United States)

Marraccini, Philip J; Riza, Nabeel A

2011-12-01

Presented is a novel design of a multimode laser beam analyzer using a digital micromirror device (DMD) and an electronically controlled variable focus lens (ECVFL) that serve as the digital and analog agile optics, respectively. The proposed analyzer is a broadband laser characterization instrument that uses the agile optics to smartly direct light to the required point photodetectors to enable beam measurements of minimum beam waist size, minimum waist location, divergence, and the beam propagation parameter M(2). Experimental results successfully demonstrate these measurements for a 500 mW multimode test laser beam with a wavelength of 532 nm. The minimum beam waist, divergence, and M(2) experimental results for the test laser are found to be 257.61 μm, 2.103 mrad, 1.600 and 326.67 μm, 2.682 mrad, 2.587 for the vertical and horizontal directions, respectively. These measurements are compared to a traditional scan method and the results of the beam waist are found to be within error tolerance of the demonstrated instrument.

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

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

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...

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....

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.

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

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.

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.

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

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.

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.

Spectral confocal reflection microscopy using a white light source

Science.gov (United States)

Booth, M.; Juškaitis, R.; Wilson, T.

2008-08-01

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

Simulation with Python on transverse modes of the symmetric confocal resonator

Science.gov (United States)

Wang, Qing Hua; Qi, Jing; Ji, Yun Jing; Song, Yang; Li, Zhenhua

2017-08-01

Python is a popular open-source programming language that can be used to simulate various optical phenomena. We have developed a suite of programs to help teach the course of laser principle. The complicated transverse modes of the symmetric confocal resonator can be visualized in personal computers, which is significant to help the students understand the pattern distribution of laser resonator.

Nano-displacement measurement based on virtual pinhole confocal method

International Nuclear Information System (INIS)

Li, Long; Kuang, Cuifang; Xue, Yi; Liu, Xu

2013-01-01

A virtual pinhole confocal system based on charge-coupled device (CCD) detection and image processing techniques is built to measure axial displacement with 10 nm resolution, preeminent flexibility and excellent robustness when facing spot drifting. Axial displacement of the sample surface is determined by capturing the confocal laser spot using a CCD detector and quantifying the energy collected by programmable virtual pinholes. Experiments indicate an applicable measuring range of 1000 nm (Gaussian fitting r = 0.9902) with a highly linear range of 500 nm (linear fitting r = 0.9993). A concentric subtraction algorithm is introduced to further enhance resolution. Factors affecting measuring precision, sensitivity and signal-to-noise ratio are discussed using theoretical deductions and diffraction simulations. The virtual pinhole technique has promising applications in surface profiling and confocal imaging applications which require easily-customizable pinhole configurations. (paper)

Assessment of Multiple Scattering Errors of Laser Diffraction Instruments

National Research Council Canada - National Science Library

Strakey, Peter

2003-01-01

The accuracy of two commercial laser diffraction instruments was compared under conditions of multiple scattering designed to simulate the high droplet number densities encountered in liquid propellant rocket combustors...

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.

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.

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.

Confocal Raman microspectroscopy

International Nuclear Information System (INIS)

Puppels, G.J.

1991-01-01

Raman spectroscopy is a technique that provides detailed structural information about molecules studied. In the field of molecular biophysics it has been extensively used for characterization of nucleic acids and proteins and for investigation of interactions between these molecules. It was felt that this technique would have great potential if it could be applied for in situ study of these molecules and their interactions, at the level of single living cell or a chromosome. To make this possible a highly sensitive confocal Raman microspectrometer (CRM) was developed. The instrument is described in detail in this thesis. It incorporates a number of recent technological developments. First, it employs a liquid nitrogen cooled CCD-camera. This type of detector, first used in astronomy, is the ultimate detector for Raman spectroscopy because it combines high quantum efficiency light detection with photon-noise limited operation. Second, an important factor in obtaining a high signal throughput of the spectrometer was the development of a new type of Raman notch filter. In the third place, the confocal detection principle was applied in the CRM. This limits the effective measuring volume to 3 . (author). 279 refs., 48 figs., 11 tabs

Ex Vivo (Fluorescence) Confocal Microscopy in Surgical Pathology: State of the Art.

Science.gov (United States)

Ragazzi, Moira; Longo, Caterina; Piana, Simonetta

2016-05-01

First developed in 1957, confocal microscopy is a powerful imaging tool that can be used to obtain near real-time reflected light images of untreated human tissue with nearly histologic resolution. Besides its research applications, in the last decades, confocal microscopy technology has been proposed as a useful device to improve clinical diagnosis, especially in ophthalmology, dermatology, and endomicroscopy settings, thanks to advances in instrument development. Compared with the wider use of the in vivo tissue assessment, ex vivo applications of confocal microscopy are not fully explored. A comprehensive review of the current literature was performed here, focusing on the reliable applications of ex vivo confocal microscopy in surgical pathology and on some potential evolutions of this new technique from pathologists' viewpoint.

Depth-variant blind restoration with pupil-phase constraints for 3D confocal microscopy

International Nuclear Information System (INIS)

Hadj, Saima Ben; Blanc-Féraud, Laure; Engler, Gilbert

2013-01-01

Three-dimensional images of confocal laser scanning microscopy suffer from a depth-variant blur, due to refractive index mismatch between the different mediums composing the system as well as the specimen, leading to optical aberrations. Our goal is to develop an image restoration method for 3D confocal microscopy taking into account the blur variation with depth. The difficulty is that optical aberrations depend on the refractive index of the biological specimen. The depth-variant blur function or the Point Spread Function (PSF) is thus different for each observation. A blind or semi-blind restoration method needs to be developed for this system. For that purpose, we use a previously developed algorithm for the joint estimation of the specimen function (original image) and the 3D PSF, the continuously depth-variant PSF is approximated by a convex combination of a set of space-invariant PSFs taken at different depths. We propose to add to that algorithm a pupil-phase constraint for the PSF estimation, given by the the optical instrument geometry. We thus define a blind estimation algorithm by minimizing a regularized criterion in which we integrate the Gerchberg-Saxton algorithm allowing to include these physical constraints. We show the efficiency of this method relying on some numerical tests

Parallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging.

Science.gov (United States)

Zhao, Ming; Li, Yu; Peng, Leilei

2014-05-05

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

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)

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.

Calibration of the geometrical characteristics of areal surface topography measuring instruments

International Nuclear Information System (INIS)

Giusca, C L; Leach, R K; Helery, F; Gutauskas, T

2011-01-01

The use of areal surface topography measuring instruments has increased significantly over the past ten years as industry starts to embrace the use of surface structuring to affect the function of a component. This has led to a range of areal surface topography measuring instruments being developed and becoming available commercially. For such instruments to be used as part of quality control during production, it is essential for them to be calibrated according to international standards. The ISO 25178 suite of specification standards on areal surface topography measurement presents a series of tests that can be used to calibrate the metrological characteristics of an areal surface topography measuring instrument. Calibration artefacts and test procedures have been developed that are compliant with ISO 25178. The material measures include crossed gratings, resolution artefacts and pseudorandom surfaces. Traceability is achieved through the NPL Areal Instrument - a primary stylus-based instrument that uses laser interferometers to measure the displacement of the stylus tip. Good practice guides on areal calibration have also been drafted for stylus instruments, coherence scanning interferometers, scanning confocal microscopes and focus variation instruments.

A compact narrow-linewidth laser with a low-Q monolithic cavity

International Nuclear Information System (INIS)

Peng, Yu

2013-01-01

We demonstrate an approach to narrowing the linewidth of a diode laser to around 15×10 3 Hz with a compact setup of confocal and parallel monolithic Fabry–Perot cavities (MFCs). Resonances of the confocal and parallel MFCs with low finesse are obtained. Diode lasers with optical feedback from confocal and parallel monolithic MFCs are demonstrated. The frequency could be tuned 80×10 6 Hz by changing the grating position of the external cavity diode laser based on the confocal MFC, and 100×10 6 Hz by tuning the temperature of the plane MFC over 0.02 ° C for the external cavity diode laser based on the parallel MFC. (paper)

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.

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.

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.

A multi-axis confocal rheoscope for studying shear flow of structured fluids

KAUST Repository

Lin, Neil Y. C.

2014-03-01

We present a new design for a confocal rheoscope that enables uniform uniaxial or biaxial shear. The design consists of two precisely positioned parallel plates with a gap that can be adjusted down to 2 ±0.1 μm, allowing for the exploration of confinement effects. By using our shear cell in conjunction with a biaxial force measurement device and a high-speed confocal microscope, we are able to measure the real-time biaxial stress while simultaneously imaging the material three-dimensional structure. We illustrate the importance of the instrument capabilities by discussing the applications of this instrument in current and future research topics in colloidal suspensions. © 2014 AIP Publishing LLC.

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)

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)

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.

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.

Local high precision 3D measurement based on line laser measuring instrument

Science.gov (United States)

Zhang, Renwei; Liu, Wei; Lu, Yongkang; Zhang, Yang; Ma, Jianwei; Jia, Zhenyuan

2018-03-01

In order to realize the precision machining and assembly of the parts, the geometrical dimensions of the surface of the local assembly surfaces need to be strictly guaranteed. In this paper, a local high-precision three-dimensional measurement method based on line laser measuring instrument is proposed to achieve a high degree of accuracy of the three-dimensional reconstruction of the surface. Aiming at the problem of two-dimensional line laser measuring instrument which lacks one-dimensional high-precision information, a local three-dimensional profile measuring system based on an accurate single-axis controller is proposed. First of all, a three-dimensional data compensation method based on spatial multi-angle line laser measuring instrument is proposed to achieve the high-precision measurement of the default axis. Through the pretreatment of the 3D point cloud information, the measurement points can be restored accurately. Finally, the target spherical surface is needed to make local three-dimensional scanning measurements for accuracy verification. The experimental results show that this scheme can get the local three-dimensional information of the target quickly and accurately, and achieves the purpose of gaining the information and compensating the error for laser scanner information, and improves the local measurement accuracy.

Development of a versatile laser light scattering instrument

Science.gov (United States)

Meyer, William V.; Ansari, Rafat R.

1990-10-01

A versatile laser light scattering (LLS) instrument is developed for use in microgravity to measure microscopic particles of 30 A to above 3 microns. Since it is an optical technique, LLS does not affect the sample being studied. A LLS instrument built from modules allows several configurations, each optimized for a particular experiment. The multiangle LLS instrument can be mounted in the rack in the Space Shuttle and on Space Station Freedom. It is possible that a Space Shuttle glove-box and a lap-top computer containing a correlator card can be used to perform a number of experiments and to demonstrate the technology needed for more elaborate investigations. This offers simple means of flying a great number of experiments without the additional requirements of full-scale flight hardware experiments.

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.

Site-specific confocal fluorescence imaging of biological microstructures in a turbid medium

International Nuclear Information System (INIS)

Saloma, Caesar; Palmes-Saloma, Cynthia; Kondoh, Hisato

1998-01-01

Normally transparent biological structures in a turbid medium are imaged using a laser confocal microscope and multiwavelength site-specific fluorescence labelling. The spatial filtering capability of the detector pinhole in the confocal microscope limits the number of scattered fluorescent photons that reach the photodetector. Simultaneous application of different fluorescent markers on the same sample site minimizes photobleaching by reducing the excitation time for each marker. A high-contrast grey-level image is also produced by summing confocal images of the same site taken at different fluorescence wavelengths. Monte Carlo simulations are performed to obtain the quantitative behaviour of confocal fluorescence imaging in turbid media. Confocal images of the following samples were also obtained: (i) 15 μm diameter fluorescent spheres placed 1.16 mm deep beneath an aqueous suspension of 0.0823 μm diameter polystyrene latex spheres, and (ii) hindbrain of a whole-mount mouse embryo (age 10 days) that was stained to fluoresce at 515 nm and 580 nm peak wavelengths. Expression of RNA transcripts of a gene within the embryo hindbrain was detected by a fluorescence-based whole-mount in situ hybridization procedure that we recently tested. (author)

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.

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...

In vivo confocal Raman spectroscopy of the human cornea.

Science.gov (United States)

Bauer, N J; Hendrikse, F; March, W F

1999-07-01

To investigate the feasibility of a confocal Raman spectroscopic technique for the noninvasive assessment of corneal hydration in vivo in two legally blind subjects. A laser beam (632.8 nm; 15 mJ) was maintained on the cornea by using a microscope objective lens (x25 magnification, NA = 0.5, f = 10 mm) both for focusing the incident light as well as collecting the Raman backscattered light, in a 180 degrees backscatter configuration. An optical fiber, acting as the confocal pinhole for elimination of light from out-of-focus places, was coupled to a spectrometer that dispersed the collected light onto a sensitive array detector for rapid spectral data acquisition over a range from 2,890 to 3,590/cm(-1). Raman spectra were recorded from the anterior 100-150 microm of the cornea over a period before and after topical application of a mild dehydrating solution. The ratio between the amplitudes of the signals at 3,400/cm(-1) (OH-vibrational mode of water) and 2,940/cm(-1) (CH-vibrational mode of proteins) was used as a measure for corneal hydration. High signal-to-noise ratio (SNR = 25) Raman spectra were obtained from the human corneas by using 15 mJ of laser light energy. Qualitative changes in the hydration of the anteriormost part of the corneas could be observed as a result of the dehydrating agent. With adequate improvements in system safety, confocal Raman spectroscopy could potentially be applied clinically as a noninvasive tool for the assessment of corneal hydration in vivo.

Cavity-enhanced quantum-cascade laser-based instrument for carbon monoxide measurements.

Science.gov (United States)

Provencal, Robert; Gupta, Manish; Owano, Thomas G; Baer, Douglas S; Ricci, Kenneth N; O'Keefe, Anthony; Podolske, James R

2005-11-01

An autonomous instrument based on off-axis integrated cavity output spectroscopy has been developed and successfully deployed for measurements of carbon monoxide in the troposphere and tropopause onboard a NASA DC-8 aircraft. The instrument (Carbon Monoxide Gas Analyzer) consists of a measurement cell comprised of two high-reflectivity mirrors, a continuous-wave quantum-cascade laser, gas sampling system, control and data-acquisition electronics, and data-analysis software. CO measurements were determined from high-resolution CO absorption line shapes obtained by tuning the laser wavelength over the R(7) transition of the fundamental vibration band near 2172.8 cm(-1). The instrument reports CO mixing ratio (mole fraction) at a 1-Hz rate based on measured absorption, gas temperature, and pressure using Beer's Law. During several flights in May-June 2004 and January 2005 that reached altitudes of 41,000 ft (12.5 km), the instrument recorded CO values with a precision of 0.2 ppbv (1-s averaging time) and an accuracy limited by the reference CO gas cylinder (uncertainty < 1.0%). Despite moderate turbulence and measurements of particulate-laden airflows, the instrument operated consistently and did not require any maintenance, mirror cleaning, or optical realignment during the flights.

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.

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

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.

Imaging subsurface damage of grinded fused silica optics by confocal fluorescence microscopy

International Nuclear Information System (INIS)

Neauport, J.; Cormont, P.; Destribats, J.; Legros, P.; Ambard, C.

2009-01-01

We report an experimental investigation of fluorescence confocal microscopy as a tool to measure subsurface damage on grinded fused silica optics. Confocal fluorescence microscopy was performed with an excitation at the wavelength of 405 nm on fixed abrasive diamond grinded fused silica samples. We detail the measured fluorescence spectrums and compare them to those of oil based coolants and grinding slurries. We evidence that oil based coolant used in diamond grinding induces a fluorescence that marks the subsurface damages and eases its observation. Such residual traces might also be involved in the laser damage process. (authors)

RELIABILITY OF CONFOCAL MICROSCOPY SPECTRAL IMAGING SYSTEMS: USE OF MULTISPECTRAL BEADS

Science.gov (United States)

Background: There is a need for a standardized, impartial calibration, and validation protocol on confocal spectral imaging (CSI) microscope systems. To achieve this goal, it is necessary to have testing tools to provide a reproducible way to evaluate instrument performance. ...

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...

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...

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...

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

Confocal detection of Rayleigh scattering for residual stress measurement in chemically tempered glass

Energy Technology Data Exchange (ETDEWEB)

Hödemann, S., E-mail: siim.hodemann@ut.ee; Möls, P.; Kiisk, V.; Saar, R.; Kikas, J. [Institute of Physics, University of Tartu, Wilhelm Ostwald st., Tartu 50411 (Estonia); Murata, T. [Nippon Electric Glass Co., 7-1 Seiran 2-chome, Otsu-shi, Shiga 520-8639 (Japan)

2015-12-28

A new optical method is presented for evaluation of the stress profile in chemically tempered (chemically strengthened) glass based on confocal detection of scattered laser beam. Theoretically, a lateral resolution of 0.2 μm and a depth resolution of 0.6 μm could be achieved by using a confocal microscope with high-NA immersion objective. The stress profile in the 250 μm thick surface layer of chemically tempered lithium aluminosilicate glass was measured with a high spatial resolution to illustrate the capability of the method. The confocal method is validated using transmission photoelastic and Na{sup +} ion concentration profile measurement. Compositional influence on the stress-optic coefficient is calculated and discussed. Our method opens up new possibilities for three-dimensional scattered light tomography of mechanical imaging in birefringent materials.

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.

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®

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.

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.

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.

Electrophoretic Detection and Confocal Microscopic Imaging of Tyrosine Nitrated Proteins in Plant Tissue.

Science.gov (United States)

Arora, Dhara; Singh, Neha; Bhatla, Satish C

2018-01-01

Tyrosine nitrated proteins can be detected in plant cells electrophoretically and their distribution can be monitored by confocal laser scanning microscopy (CLSM) imaging. One-dimensional polyacrylamide gel electrophoresis (1D PAGE) followed by Western blotting using polyclonal antibody against 3-nitrotyrosine residues enables detection of tyrosine nitrated proteins in plant cells. Here we describe detection of tyrosine nitrated proteins in the homogenates derived from sunflower (Helianthus annuus L.) seedling cotyledons. Total soluble proteins obtained from tissue homogenates are resolved using vertical gel electrophoresis followed by their electrophoretic transfer on to a microporous membrane support for immunodetection. Spatial distribution of tyrosine nitrated proteins can be visualized using an antibody against 3-nitrotyrosine residues. Immunofluorescent localization is performed by cutting 7 μm thick wax sections of tissue followed by incubation in primary anti-nitrotyrosine antibody (dilution 1:200) and secondary Cy-3 labeled anti-rabbit IgG antibody (dilution 1:1500). Confocal laser scanning microscopy analysis is undertaken using argon lasers (ex: 530-550 nm and em: 570 nm) at pinhole 1. Modulation in the abundance and spatial localization of tyrosine nitrated proteins in plant tissues can be monitored using these techniques.

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.

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.

The laser absorption spectrometer - A new remote sensing instrument for atmospheric pollution monitoring

Science.gov (United States)

Shumate, M. S.

1974-01-01

An instrument capable of remotely monitoring trace atmospheric constituents is described. The instrument, called a laser absorption spectrometer, can be operated from an aircraft or spacecraft to measure the concentration of selected gases in three dimensions. This device will be particularly useful for rapid determination of pollutant levels in urban areas.

Confocal Raman spectrocopy for the analysis of nail polish evidence.

Science.gov (United States)

López-López, Maria; Vaz, Joana; García-Ruiz, Carmen

2015-06-01

Nail polishes are cosmetic paints that may be susceptible of forensic analysis offering useful information to assist in a crime reconstruction. Although the nail polish appearance could allow a quick visual identification of the sample, this analysis is subjected to the perception and subjective interpretation of the forensic examiner. The chemical analysis of the nail polishes offers great deal of information not subjected to analyst interpretation. Confocal Raman spectroscopy is a well-suited technique for the analysis of paints due to its non-invasive and non-destructive nature and its ability to supply information about the organic and inorganic components of the sample. In this work, 77 regular and gel nail polishes were analyzed with confocal Raman spectroscopy using two laser wavelengths (532 and 780 nm). The sample behavior under the two laser wavelengths and the differences in the spectra taken at different points of the sample were studied for each nail polish. Additionally, the spectra obtained for all the nail polishes were visually compared. The results concluded that the longer laser wavelength prevents sample burning and fluorescence effects; the similarity among the spectra collected within the sample is not directly related with the presence of glitter particles; and 64% of the samples analyzed showed a characteristic spectrum. Additionally, the use of confocal Raman spectroscopy for the forensic analysis of nail polishes evidence in the form of flakes or smudges on different surfaces were studied. The results showed that both types of evidence can be analyzed by the technique. Also, two non-invasive sampling methods for the collection of the evidence from the nails of the suspect or the victim were proposed: (i) to use acetone-soaked cotton swabs to remove the nail varnishes and (ii) to scrape the nail polish from the nail with a blade. Both approaches, each exhibiting advantages and drawbacks in terms of transport and handling were appropriate

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.

Atmospheric effects on laser eye safety and damage to instrumentation

Science.gov (United States)

Zilberman, Arkadi; Kopeika, Natan S.

2017-10-01

Electro-optical sensors as well as unprotected human eyes are extremely sensitive to laser radiation and can be permanently damaged from direct or reflected beams. Laser detector/eye hazard depends on the interaction between the laser beam and the media in which it traverses. The environmental conditions including terrain features, atmospheric particulate and water content, and turbulence, may alter the laser's effect on the detector/eye. It is possible to estimate the performance of an electro-optical system as long as the atmospheric propagation of the laser beam can be adequately modeled. More recent experiments and modeling of atmospheric optics phenomena such as inner scale effect, aperture averaging, atmospheric attenuation in NIR-SWIR, and Cn2 modeling justify an update of previous eye/detector safety modeling. In the present work, the influence of the atmospheric channel on laser safety for personnel and instrumentation is shown on the basis of theoretical and experimental data of laser irradiance statistics for different atmospheric conditions. A method for evaluating the probability of damage and hazard distances associated with the use of laser systems in a turbulent atmosphere operating in the visible and NIR-SWIR portions of the electromagnetic spectrum is presented. It can be used as a performance prediction model for directed energy engagement of ground-based or air-based systems.

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.

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.

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.

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.

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.

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

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.

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

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.

Fluorescence confocal microscopy for pathologists.

Science.gov (United States)

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

2014-03-01

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

Design considerations of a real-time clinical confocal microscope

Science.gov (United States)

Masters, Barry R.

1991-06-01

A real-time clinical confocal light microscope provides the ophthalmologist with a new tool for the observation of the cornea and the ocular lens. In addition, the ciliary body, the iris, and the sclera can be observed. The real-time light microscopic images have high contrast and resolution. The transverse resolution is about one half micron and the range resolution is one micron. The following observations were made with visible light: corneal epithelial cells, wing cells, basal cells, Bowman's membrane, nerve fibers, basal lamina, fibroblast nuclei, Descemet's membrane, endothelial cells. Observation of the in situ ocular lens showed lens capsule, lens epithelium, lens fibrils, the interior of lens fibrils. The applications of the confocal microscope include: eye banking, laser refractive surgery, observation of wound healing, observation of the iris, the sciera, the ciliary body, the ocular lens, and the intraocular lens. Digital image processing can produce three-dimensional reconstructions of the cornea and the ocular lens.

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

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

Confocal fluorescence microscopy for minimal-invasive tumor diagnosis

International Nuclear Information System (INIS)

Zenzinger, M.; Bille, J.

2000-01-01

The goal of the project ''stereotactic laser-neurosurgery'' is the development of a system for careful and minimal-invasive resection of brain tumors with ultrashort laser pulses through a thin probe. A confocal laser-scanning-microscope is integrated in the probe. In this paper, the simulation of its optical properties by a laboratory setup and the expansion by the ability for fluorescence microscopy are reported. For a valuation of the imaging properties, the point-spread-function in three dimensions and the axial depth-transfer-function were measured and thus, among other things, the resolving power and the capacity for depth discrimination were analysed. The microscope will enable intra-operative detection of tumor cells by the method of immunofluorescence. As a first model of the application in the brain, cell cultures, that fluorescein-labelled antibodies were bound to specifically, were used in this work. Due to the fluorescence signal, it was possible to detect and identify clearly the areas that had been marked in this manner, proving the suitability of the setup for minimal-invasive tumor diagnosis. (orig.)

3D Volumetric Analysis of Fluid Inclusions Using Confocal Microscopy

Science.gov (United States)

Proussevitch, A.; Mulukutla, G.; Sahagian, D.; Bodnar, B.

2009-05-01

Fluid inclusions preserve valuable information regarding hydrothermal, metamorphic, and magmatic processes. The molar quantities of liquid and gaseous components in the inclusions can be estimated from their volumetric measurements at room temperatures combined with knowledge of the PVTX properties of the fluid and homogenization temperatures. Thus, accurate measurements of inclusion volumes and their two phase components are critical. One of the greatest advantages of the Laser Scanning Confocal Microscopy (LSCM) in application to fluid inclsion analsyis is that it is affordable for large numbers of samples, given the appropriate software analysis tools and methodology. Our present work is directed toward developing those tools and methods. For the last decade LSCM has been considered as a potential method for inclusion volume measurements. Nevertheless, the adequate and accurate measurement by LSCM has not yet been successful for fluid inclusions containing non-fluorescing fluids due to many technical challenges in image analysis despite the fact that the cost of collecting raw LSCM imagery has dramatically decreased in recent years. These problems mostly relate to image analysis methodology and software tools that are needed for pre-processing and image segmentation, which enable solid, liquid and gaseous components to be delineated. Other challenges involve image quality and contrast, which is controlled by fluorescence of the material (most aqueous fluid inclusions do not fluoresce at the appropriate laser wavelengths), material optical properties, and application of transmitted and/or reflected confocal illumination. In this work we have identified the key problems of image analysis and propose some potential solutions. For instance, we found that better contrast of pseudo-confocal transmitted light images could be overlayed with poor-contrast true-confocal reflected light images within the same stack of z-ordered slices. This approach allows one to narrow

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.

Implementação de um sistema de eletroforese capilar com detecção de fluorescência induzida por laser

Directory of Open Access Journals (Sweden)

Santos Marcia R.

2000-01-01

Full Text Available A capillary electrophoresis system using laser induced fluorescence detection is described. A Raman system equipped with a microscope has been used to focus the laser beam on the capillary giving a lateral resolution of 1.5 mm. The fluorescence signal of the analyte (ZnPcTS - tetrasulfonated zinc-phthalocyanine was collected by the microscope objectives and analysed by a monochromator with confocal characteristics equipped with a CCD detector. Electropherograms obtained with this system were compared to those obtained on a commercial instrument, showing that the described system presents a lower detection limit and better resolution.

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.

Multi-spectral confocal microendoscope for in-vivo imaging

Science.gov (United States)

Rouse, Andrew Robert

The concept of in-vivo multi-spectral confocal microscopy is introduced. A slit-scanning multi-spectral confocal microendoscope (MCME) was built to demonstrate the technique. The MCME employs a flexible fiber-optic catheter coupled to a custom built slit-scan confocal microscope fitted with a custom built imaging spectrometer. The catheter consists of a fiber-optic imaging bundle linked to a miniature objective and focus assembly. The design and performance of the miniature objective and focus assembly are discussed. The 3mm diameter catheter may be used on its own or routed though the instrument channel of a commercial endoscope. The confocal nature of the system provides optical sectioning with 3mum lateral resolution and 30mum axial resolution. The prism based multi-spectral detection assembly is typically configured to collect 30 spectral samples over the visible chromatic range. The spectral sampling rate varies from 4nm/pixel at 490nm to 8nm/pixel at 660nm and the minimum resolvable wavelength difference varies from 7nm to 18nm over the same spectral range. Each of these characteristics are primarily dictated by the dispersive power of the prism. The MCME is designed to examine cellular structures during optical biopsy and to exploit the diagnostic information contained within the spectral domain. The primary applications for the system include diagnosis of disease in the gastro-intestinal tract and female reproductive system. Recent data from the grayscale imaging mode are presented. Preliminary multi-spectral results from phantoms, cell cultures, and excised human tissue are presented to demonstrate the potential of in-vivo multi-spectral imaging.

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

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.

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.

Analysis of cell-tissue grafts under weightless conditions using confocal fluorescence microscopy

Science.gov (United States)

Volova, L. T.; Milyakova, M. N.; Rossinskaya, V. V.; Boltovskaya, V. V.; Kulagina, L. N.; Kurganskaya, L. V.; Timchenko, P. E.; Timchenko, E. V.; Zherdeva Taskina, Larisa A.

2015-03-01

The research results of monitoring of viable cells in a cellular-tissue graft using confocal laser fluorescence microscopy at 488 nm and 561 nm with the use of fluorophore propidium iodide (propidium iodide, PI Sigma Aldrich USA) are presented. The processing of the received images was carried out using the software ANDOR. It is experimentally shown that the method of confocal fluorescence microscopy is one of the informational methods for detecting cells populated in a 3-D bio-carrier with a resolution of at least 400 nm. Analysis of the received micrographs suggests that the cells that were in a bio-carrier for 30 days in a synchronous ground-based experiment retained their viability compared to a similar space-based experiment in which the cells were hardly detected in a bio-carrier.

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.

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.

Influence of instrument conditions on the evaporation behavior of uranium dioxide with UV laser-assisted atom probe tomography

International Nuclear Information System (INIS)

2015-01-01

Atom probe tomography (APT) provides the ability to detect subnanometer chemical variations spatially with high accuracy. Due to its ability to spatially characterize chemistry in non-conducting materials, such as oxides, provides the opportunity to characterize stoichiometry, which strongly is tied to material performance. However, accuracy has been correlated with instrument run parameters. A systematic study of the effect of laser energy, temperature, and detection rate is performed on the evaporation behavior of a model oxide, uranium dioxide (UO 2 ). Modifying the detection rate and temperature did not affect its evaporation behavior as laser energy. It was discovered that three laser evaporation regimes are present in UO 2 . Very low laser energy produces a behavior similar to DC-field evaporation, moderate laser energy produces the desired laser assisted field evaporation and high laser energy produces thermal effects in the evaporation behavior. Laser energy had the greatest impact on evaporation and the optimal instrument condition for UO 2 was determined to be 50K, 10 pJ laser energy, 0.3% detection rate, and a 100 kHz repetition rate. These conditions provide the best combination of mass resolution, accurate stoichiometry, and evaporation behavior.

Transverse mode selection in a monolithic microchip laser

CSIR Research Space (South Africa)

Naidoo, Darryl

2011-11-01

Full Text Available selection in a monolithic microchip laser Darryl Naidooa,b, Thomas Godinc, Michael Fromagerc, Emmanuel Cagniotc, Nicolas Passillyd, Andrew Forbesa,b and Kamel A?t-Ameurc1 a:CSIR National Laser Centre, P. O. Box 395, Pretoria 0001, South Africa b.... Lett. 77 (2000) 34-36. [14] W. Zhao, J. Tan and L. Qui, ?Improvement of confocal microscope performance by shaped annular beam and heterodyne confocal techniques,? Optik 116 (2005) 111-117. [15] T. Shiina, K. Yoshida, M. Ito and Y. Okamura, ?Long...

Development of Mechanical Sealing and Laser Welding Technology to Instrument Thermocouple for Nuclear Fuel Test Rod

Energy Technology Data Exchange (ETDEWEB)

Joung, Chang-Young; Ahn, Sung-Ho; Hong, Jin-Tae; Kim, Ka-Hye; Huh, Sung-Ho [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2015-05-15

Zircaloy-4 of the nuclear fuel test rod, AISI 316L of the mechanical sealing parts, and the MI (mineral insulated) cable at a thermocouple instrumentation are hetero-metals, and are difficult to weld to dissimilar materials. Therefore, a mechanical sealing method to instrument the thermocouple should be conducted using two kinds of sealing process as follows: One is a mechanical sealing process using Swagelok, which is composed of sealing components that consists of an end-cap, a seal tube, a compression ring and a Swagelok nut. The other is a laser welding process used to join a seal tube, and an MI cable, which are made of the same material. The mechanical sealing process should be sealed up with the mechanical contact compressed by the strength forced between a seal tube and an end-cap, and the laser welding process should be conducted to have no defects on the sealing area between a seal tube and an MI cable. Therefore, the mechanical sealing and laser welding techniques need to be developed to accurately measure the centerline temperature of the nuclear fuel test rod in an experimental reactor. The mechanical sealing and laser welding tests were conducted to develop the thermocouple instrumentation techniques for the nuclear fuel test rod. The optimum torque value of a Swagelok nut to seal the mechanical sealing part between the end-cap and seal tube was established through various torque tests using a torque wrench. The optimum laser welding conditions to seal the welding part between a seal tube and an MI cable were obtained through various welding tests using a laser welding system.

Development of Mechanical Sealing and Laser Welding Technology to Instrument Thermocouple for Nuclear Fuel Test Rod

International Nuclear Information System (INIS)

Joung, Chang-Young; Ahn, Sung-Ho; Hong, Jin-Tae; Kim, Ka-Hye; Huh, Sung-Ho

2015-01-01

Zircaloy-4 of the nuclear fuel test rod, AISI 316L of the mechanical sealing parts, and the MI (mineral insulated) cable at a thermocouple instrumentation are hetero-metals, and are difficult to weld to dissimilar materials. Therefore, a mechanical sealing method to instrument the thermocouple should be conducted using two kinds of sealing process as follows: One is a mechanical sealing process using Swagelok, which is composed of sealing components that consists of an end-cap, a seal tube, a compression ring and a Swagelok nut. The other is a laser welding process used to join a seal tube, and an MI cable, which are made of the same material. The mechanical sealing process should be sealed up with the mechanical contact compressed by the strength forced between a seal tube and an end-cap, and the laser welding process should be conducted to have no defects on the sealing area between a seal tube and an MI cable. Therefore, the mechanical sealing and laser welding techniques need to be developed to accurately measure the centerline temperature of the nuclear fuel test rod in an experimental reactor. The mechanical sealing and laser welding tests were conducted to develop the thermocouple instrumentation techniques for the nuclear fuel test rod. The optimum torque value of a Swagelok nut to seal the mechanical sealing part between the end-cap and seal tube was established through various torque tests using a torque wrench. The optimum laser welding conditions to seal the welding part between a seal tube and an MI cable were obtained through various welding tests using a laser welding system

ALADIN: an atmospheric laser Doppler wind lidar instrument for wind velocity measurements from space

Science.gov (United States)

Krawczyk, R.; Ghibaudo, JB.; Labandibar, JY.; Willetts, D.; Vaughan, M.; Pearson, G.; Harris, M.; Flamant, P. H.; Salamitou, P.; Dabas, A.; Charasse, R.; Midavaine, T.; Royer, M.; Heimel, H.

2018-04-01

This paper, "ALADIN: an atmospheric laser Doppler wind lidar instrument for wind velocity measurements from space," was presented as part of International Conference on Space Optics—ICSO 1997, held in Toulouse, France.

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.

Control of the positional relationship between a sample collection instrument and a surface to be analyzed during a sampling procedure using a laser sensor

Science.gov (United States)

Van Berkel, Gary J [Clinton, TN; Kertesz, Vilmos [Knoxville, TN

2012-02-21

A system and method utilizes distance-measuring equipment including a laser sensor for controlling the collection instrument-to-surface distance during a sample collection process for use, for example, with mass spectrometric detection. The laser sensor is arranged in a fixed positional relationship with the collection instrument, and a signal is generated by way of the laser sensor which corresponds to the actual distance between the laser sensor and the surface. The actual distance between the laser sensor and the surface is compared to a target distance between the laser sensor and the surface when the collection instrument is arranged at a desired distance from the surface for sample collecting purposes, and adjustments are made, if necessary, so that the actual distance approaches the target distance.

Metrological characterization methods for confocal chromatic line sensors and optical topography sensors

Science.gov (United States)

Seppä, Jeremias; Niemelä, Karri; Lassila, Antti

2018-05-01

The increasing use of chromatic confocal technology for, e.g. fast, in-line optical topography, and measuring thickness, roughness and profiles implies a need for the characterization of various aspects of the sensors. Single-point, line and matrix versions of chromatic confocal technology, encoding depth information into wavelength, have been developed. Of these, line sensors are particularly suitable for in-line process measurement. Metrological characterization and development of practical methods for calibration and checking is needed for new optical methods and devices. Compared to, e.g. tactile methods, optical topography measurement techniques have limitations related to light wavelength and coherence, optical properties of the sample including reflectivity, specularity, roughness and colour, and definition of optical versus mechanical surfaces. In this work, metrological characterization methods for optical line sensors were developed for scale magnification and linearity, sensitivity to sample properties, and dynamic characteristics. An accurate depth scale calibration method using a single prototype groove depth sample was developed for a line sensor and validated with laser-interferometric sample tracking, attaining (sub)micrometre level or better than 0.1% scale accuracy. Furthermore, the effect of different surfaces and materials on the measurement and depth scale was studied, in particular slope angle, specularity and colour. In addition, dynamic performance, noise, lateral scale and resolution were measured using the developed methods. In the case of the LCI1200 sensor used in this study, which has a 11.3 mm  ×  2.8 mm measurement range, the instrument depth scale was found to depend only minimally on sample colour, whereas measuring steeply sloped specular surfaces in the peripheral measurement area, in the worst case, caused a somewhat larger relative sample-dependent change (1%) in scale.

High resolution 3D confocal microscope imaging of volcanic ash particles.

Science.gov (United States)

Wertheim, David; Gillmore, Gavin; Gill, Ian; Petford, Nick

2017-07-15

We present initial results from a novel high resolution confocal microscopy study of the 3D surface structure of volcanic ash particles from two recent explosive basaltic eruptions, Eyjafjallajökull (2010) and Grimsvötn (2011), in Iceland. The majority of particles imaged are less than 100μm in size and include PM 10 s, known to be harmful to humans if inhaled. Previous studies have mainly used 2D microscopy to examine volcanic particles. The aim of this study was to test the potential of 3D laser scanning confocal microscopy as a reliable analysis tool for these materials and if so to what degree high resolution surface and volume data could be obtained that would further aid in their classification. First results obtained using an Olympus LEXT scanning confocal microscope with a ×50 and ×100 objective lens are highly encouraging. They reveal a range of discrete particle types characterised by sharp or concave edges consistent with explosive formation and sudden rupture of magma. Initial surface area/volume ratios are given that may prove useful in subsequent modelling of damage to aircraft engines and human tissue where inhalation has occurred. Copyright © 2017 Elsevier B.V. All rights reserved.

Multifocus confocal Raman microspectroscopy for fast multimode vibrational imaging of living cells.

Science.gov (United States)

Okuno, Masanari; Hamaguchi, Hiro-o

2010-12-15

We have developed a multifocus confocal Raman microspectroscopic system for the fast multimode vibrational imaging of living cells. It consists of an inverted microscope equipped with a microlens array, a pinhole array, a fiber bundle, and a multichannel Raman spectrometer. Forty-eight Raman spectra from 48 foci under the microscope are simultaneously obtained by using multifocus excitation and image-compression techniques. The multifocus confocal configuration suppresses the background generated from the cover glass and the cell culturing medium so that high-contrast images are obtainable with a short accumulation time. The system enables us to obtain multimode (10 different vibrational modes) vibrational images of living cells in tens of seconds with only 1 mW laser power at one focal point. This image acquisition time is more than 10 times faster than that in conventional single-focus Raman microspectroscopy.

Laser stimulation can activate autophagy in HeLa cells

International Nuclear Information System (INIS)

Wang, Yisen; Hu, Minglie; Wang, Chingyue; Lan, Bei; Cao, Youjia; He, Hao

2014-01-01

For decades, lasers have been a daily tool in most biological research for fluorescent excitation by confocal or multiphoton microscopy. More than 20 years ago, cell photodamage caused by intense laser stimulation was noticed by generating reactive oxygen species, which was then thought as the main damage effect by photons. In this study, we show that laser stimulation can induce autophagy, an important cell lysosomal pathway responding to immune stimulation and starvation, without any biochemical treatment. Two different types of laser stimulations are found to be capable of activating autophagy: continuous scanning by continuous-wave visible lasers and a short-time flash of femtosecond laser irradiation. The autophagy generation is independent from wavelength, power, and scanning duration of the visible lasers. In contrast, the power of femtosecond laser is very critical to autophagy because the multiphoton excited Ca 2+ dominates autophagy signaling. In general, we show here the different mechanisms of autophagy generation by such laser stimulation, which correspond to confocal microscopy and cell surgery, respectively. Those results can help further understanding of photodamage and autophagy signaling.

Laser stimulation can activate autophagy in HeLa cells

Energy Technology Data Exchange (ETDEWEB)

Wang, Yisen; Hu, Minglie; Wang, Chingyue [Ultrafast Laser Laboratory, Key Laboratory of Optoelectronic Information Technology (Ministry of Education), College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin (China); Lan, Bei; Cao, Youjia [Key Laboratory of Microbial Functional Genomics of Ministry of Education, College of Life Sciences, Nankai University, Tianjin (China); He, Hao, E-mail: haohe@tju.edu.cn [Ultrafast Laser Laboratory, Key Laboratory of Optoelectronic Information Technology (Ministry of Education), College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin (China); Med-X Research Institute, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai (China)

2014-10-27

For decades, lasers have been a daily tool in most biological research for fluorescent excitation by confocal or multiphoton microscopy. More than 20 years ago, cell photodamage caused by intense laser stimulation was noticed by generating reactive oxygen species, which was then thought as the main damage effect by photons. In this study, we show that laser stimulation can induce autophagy, an important cell lysosomal pathway responding to immune stimulation and starvation, without any biochemical treatment. Two different types of laser stimulations are found to be capable of activating autophagy: continuous scanning by continuous-wave visible lasers and a short-time flash of femtosecond laser irradiation. The autophagy generation is independent from wavelength, power, and scanning duration of the visible lasers. In contrast, the power of femtosecond laser is very critical to autophagy because the multiphoton excited Ca{sup 2+} dominates autophagy signaling. In general, we show here the different mechanisms of autophagy generation by such laser stimulation, which correspond to confocal microscopy and cell surgery, respectively. Those results can help further understanding of photodamage and autophagy signaling.

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

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.

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)

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.

3D Image Analysis of Geomaterials using Confocal Microscopy

Science.gov (United States)

Mulukutla, G.; Proussevitch, A.; Sahagian, D.

2009-05-01

Confocal microscopy is one of the most significant advances in optical microscopy of the last century. It is widely used in biological sciences but its application to geomaterials lingers due to a number of technical problems. Potentially the technique can perform non-invasive testing on a laser illuminated sample that fluoresces using a unique optical sectioning capability that rejects out-of-focus light reaching the confocal aperture. Fluorescence in geomaterials is commonly induced using epoxy doped with a fluorochrome that is impregnated into the sample to enable discrimination of various features such as void space or material boundaries. However, for many geomaterials, this method cannot be used because they do not naturally fluoresce and because epoxy cannot be impregnated into inaccessible parts of the sample due to lack of permeability. As a result, the confocal images of most geomaterials that have not been pre-processed with extensive sample preparation techniques are of poor quality and lack the necessary image and edge contrast necessary to apply any commonly used segmentation techniques to conduct any quantitative study of its features such as vesicularity, internal structure, etc. In our present work, we are developing a methodology to conduct a quantitative 3D analysis of images of geomaterials collected using a confocal microscope with minimal amount of prior sample preparation and no addition of fluorescence. Two sample geomaterials, a volcanic melt sample and a crystal chip containing fluid inclusions are used to assess the feasibility of the method. A step-by-step process of image analysis includes application of image filtration to enhance the edges or material interfaces and is based on two segmentation techniques: geodesic active contours and region competition. Both techniques have been applied extensively to the analysis of medical MRI images to segment anatomical structures. Preliminary analysis suggests that there is distortion in the

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.

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.

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.

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.

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.

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.

Laser-induced emission, fluorescence and Raman hybrid setup: A versatile instrument to analyze materials from cultural heritage

Science.gov (United States)

Syvilay, D.; Bai, X. S.; Wilkie-Chancellier, N.; Texier, A.; Martinez, L.; Serfaty, S.; Detalle, V.

2018-02-01

The aim of this research project was the development of a hybrid system in laboratory coupling together three analytical techniques, namely laser-induced breakdown spectroscopy (LIBS), laser-induced fluorescence (LIF) and Raman spectroscopy in a single instrument. The rationale for combining these three spectroscopies was to identify a material (molecular and elemental analysis) without any preliminary preparation, regardless of its organic or inorganic nature, on the surface and in depth, without any surrounding light interference thanks to time resolution. Such instrumentation would allow characterizing different materials from cultural heritage. A complete study on LIBS-LIF-Raman hybrid was carried out, from its conception to instrumental achievement, in order to elaborate a strategy of analysis according to the material and to be able to address conservation issues. From an instrumental point of view, condensing the three spectroscopies was achieved by using a single laser for excitation and two spectrometers (time-integrated and not time-integrated) for light collection. A parabolic mirror was used as collecting system, while three excitation sources directed through this optical system ensured the examination of a similar probe area. Two categories of materials were chosen to test the hybrid instrumentation on cultural heritage applications (copper corrosion products and wall paintings). Some examples are reported to illustrate the wealth of information provided by the hybrid, thus demonstrating its great potential to be used for cultural heritage issues. Finally, several considerations are outlined aimed at further improving the hybrid.

Development of confocal 3D micro-XRF spectrometer with dual Cr-Mo excitation

International Nuclear Information System (INIS)

Kouichi Tsuji; Kazuhiko Nakano

2007-01-01

A new 3D micro-XRF instrument based on a confocal setup using two independent poly-capillary x-ray lenses and two x-ray sources (Cr and Mo targets) was developed. A full poly-capillary x-ray lens was attached to each x-ray tube. Another half poly-capillary lens was attached to a silicon drift x-ray detector (SDD). The focal spots of the three lenses were adjusted to a common position. The depth resolutions that were evaluated by use of a 10-μm thick Au foil were approximately 90 μm for the x-ray energy of Au Lα. The effects of the dual Cr-Mo x-ray beam excitation were investigated. It was confirmed that the XRF intensity of light elements was increased by applying the Cr-target x-ray tube in a confocal configuration. In the proposed confocal configuration, 3D elemental mapping of the major elements of an amaranth seed was performed nondestructively at ambient air pressure. Each element of the seed showed different mapping images in the different depth layers. (authors)

Development of confocal 3D micro-XRF spectrometer with dual Cr-Mo excitation

Energy Technology Data Exchange (ETDEWEB)

Kouichi Tsuji [Department of Applied Chemistry, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku Osaka 558-8585 (Japan); PRESTO-JST - Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency, 4-1-8 Honcho Kawaguchi, Saitama 332-0012 (Japan); Kazuhiko Nakano [Department of Applied Chemistry, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku Osaka 558-8585 (Japan)

2007-05-15

A new 3D micro-XRF instrument based on a confocal setup using two independent poly-capillary x-ray lenses and two x-ray sources (Cr and Mo targets) was developed. A full poly-capillary x-ray lens was attached to each x-ray tube. Another half poly-capillary lens was attached to a silicon drift x-ray detector (SDD). The focal spots of the three lenses were adjusted to a common position. The depth resolutions that were evaluated by use of a 10-{mu}m thick Au foil were approximately 90 {mu}m for the x-ray energy of Au L{alpha}. The effects of the dual Cr-Mo x-ray beam excitation were investigated. It was confirmed that the XRF intensity of light elements was increased by applying the Cr-target x-ray tube in a confocal configuration. In the proposed confocal configuration, 3D elemental mapping of the major elements of an amaranth seed was performed nondestructively at ambient air pressure. Each element of the seed showed different mapping images in the different depth layers. (authors)

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.

Confocal Raman spectroscopy to trace lipstick with their smudges on different surfaces.

Science.gov (United States)

López-López, Maria; Özbek, Nil; García-Ruiz, Carmen

2014-06-01

Lipsticks are very popular cosmetic products that can be transferred by contact to different surfaces, being important forensic evidence with an intricate analysis if they are found in a crime scene. This study evaluates the use of confocal Raman microscopy at 780 nm excitation wavelength for the nondestructive identification of 49 lipsticks of different brands and colors, overcoming the lipstick fluorescence problem reported by previous works using other laser wavelengths. Although the lipsticks samples showed some fluorescence, this effect was not so intense to completely overwhelm the Raman spectra. Lipsticks smudges on twelve different surfaces commonly stained with these samples were also analyzed. In the case of the surfaces, some of them provided several bands to the smudge spectra compromising the identification of the lipstick. For these samples spectral subtraction of the interfering bands from the surface was performed. Finally, five different red lipsticks with very similar color were measured on different surfaces to evaluate the lipstick traceability with their smudges even on interfering surfaces. Although previous spectral subtraction was needed in some cases, all the smudged were linked to their corresponding lipsticks even when they are smeared on the interfering surfaces. As a consequence, confocal Raman microscopy using the 780 nm excitation laser is presented as a nondestructive powerful tool for the identification of these tricky samples. Copyright © 2014 Elsevier B.V. All rights reserved.

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.

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...

Confocal Raman microscopy

CERN Document Server

Dieing, Thomas; Hollricher, Olaf

2018-01-01

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

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.

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.

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

3D confocal imaging in CUBIC-cleared mouse heart

Energy Technology Data Exchange (ETDEWEB)

Nehrhoff, I.; Bocancea, D.; Vaquero, J.; Vaquero, J.J.; Lorrio, M.T.; Ripoll, J.; Desco, M.; Gomez-Gaviro, M.V.

2016-07-01

Acquiring high resolution 3D images of the heart enables the ability to study heart diseases more in detail. Here, the CUBIC (clear, unobstructed brain imaging cocktails and computational analysis) clearing protocol was adapted for thick mouse heart sections to increase the penetration depth of the confocal microscope lasers into the tissue. The adapted CUBIC clearing of the heart lets the antibody penetrate deeper into the tissue by a factor of five. The here shown protocol enables deep 3D highresolution image acquisition in the heart. This allows a much more accurate assessment of the cellular and structural changes that underlie heart diseases. (Author)

3D confocal imaging in CUBIC-cleared mouse heart

International Nuclear Information System (INIS)

Nehrhoff, I.; Bocancea, D.; Vaquero, J.; Vaquero, J.J.; Lorrio, M.T.; Ripoll, J.; Desco, M.; Gomez-Gaviro, M.V.

2016-01-01

Acquiring high resolution 3D images of the heart enables the ability to study heart diseases more in detail. Here, the CUBIC (clear, unobstructed brain imaging cocktails and computational analysis) clearing protocol was adapted for thick mouse heart sections to increase the penetration depth of the confocal microscope lasers into the tissue. The adapted CUBIC clearing of the heart lets the antibody penetrate deeper into the tissue by a factor of five. The here shown protocol enables deep 3D highresolution image acquisition in the heart. This allows a much more accurate assessment of the cellular and structural changes that underlie heart diseases. (Author)

An international comparison of surface texture parameters quantification on polymer artefacts using optical instruments

DEFF Research Database (Denmark)

Tosello, Guido; Haitjema, H.; Leach, R.K.

2016-01-01

An international comparison of optical instruments measuring polymer surfaces with arithmetic mean height values in the sub-micrometre range has been carried out. The comparison involved sixteen optical surface texture instruments (focus variation instruments, confocal microscopes and coherent...

Role of laser fluence in protein synthesis of cultured DRG neurons following low-level laser irradiation

Science.gov (United States)

Zheng, Liqin; Qiu, Caimin; Wang, Yuhua; Zeng, Yixiu; Yang, Hongqin; Zhang, Yanding; Xie, Shusen

2014-11-01

Low-level lasers have been used to relieve pain in clinical for many years. But the mechanism is not fully clear. In animal models, nitric oxide (NO) has been reported involving in the transmission and modulation of nociceptive signals. So the objective of this study was to establish whether low-level laser with different fluence could stimulate the production of nitric oxide synthese (NOS), which produces NO in cultured primary dorsal root ganglion neurons (DRG neurons). The primary DRG neurons were isolated from healthy Sprague Dawley rats (8-12 weeks of age) and spread on 35 mm culture dishes specially used for confocal microscopy. 24 hours after spreading, cells were irradiated with 658 nm laser for two consecutive days at the energy density of 20, 40, 60 and 80 mJ·cm-2 respectively. Control groups were not exposed to the laser, but were kept under the same conditions as the irradiated ones. The synthesis of NOS after laser irradiation was detected by immunofluorescence assay, and the changes of NOS were evaluated using confocal microscopy and Image J software. The results showed that all the laser fluence could promote the production of NOS in DRG neurons, especially the 60 mJ·cm-2 . These results demonstrated that low-level laser irradiation could modify protein synthesis in a dose- or fluence- dependent manner, and indicated that low-level laser irradiation might achieve the analgesic effect through modulation of NO production.

Instrument-independent flux units for laser Doppler perfusion monitoring assessed in a multi-device study on the renal cortex

NARCIS (Netherlands)

Petoukhova, AL; Steenbergen, W; Morales, F; Graaff, R; de Jong, ED; Elstrodt, JM; de Mul, FFM; Rakhorst, G

To investigate the feasibility of instrument-independent perfusion units for laser Doppler flowmetry, a comparison was performed of two commercial fiberoptic laser Doppler perfusion monitors measuring the same flux situation for two different types of probes. In vivo measurements were performed on

Instrument-independent flux units for laser Doppler perfusion monitoring assessed in a multi-device study on the renal cortex

NARCIS (Netherlands)

Petoukhova, Anna; Steenbergen, Wiendelt; Morales, F.; Graaff, R.; de Jong, Ed; Elstrodt, J.M.; de Mul, F.F.M.; Rakhorst, G.

2003-01-01

To investigate the feasibility of instrument-independent perfusion units for laser Doppler flowmetry, a comparison was performed of two commercial fiberoptic laser Doppler perfusion monitors measuring the same flux situation for two different types of probes. In vivo measurements were performed on

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

A Laser-Induced Fluorescence Instrument for Aircraft Measurements of Sulfur Dioxide in the Upper Troposphere and Lower Stratosphere

Science.gov (United States)

Rollins, Andrew W.; Thornberry, Troy D.; Ciciora, Steven J.; McLaughlin, Richard J.; Watts, Laurel A.; Hanisco, Thomas F.; Baumann, Esther; Giorgetta, Fabrizio R.; Bui, Thaopaul V.; Fahey, David W.

2016-01-01

This work describes the development and testing of a new instrument for in situ measurements of sulfur dioxide (SO2) on airborne platforms in the upper troposphere and lower stratosphere (UTLS). The instrument is based on the laser-induced fluorescence technique and uses the fifth harmonic of a tunable fiber-amplified semiconductor diode laser system at 1084.5 nm to excite SO2 at 216.9 nm. Sensitivity and background checks are achieved in flight by additions of SO2 calibration gas and zero air, respectively. Aircraft demonstration was performed during the NASA Volcano Plume Investigation Readiness and Gas-Phase and Aerosol Sulfur (VIRGAS) experiment, which was a series of flights using the NASA WB-57F during October 2015 based at Ellington Field and Harlingen, Texas. During these flights, the instrument successfully measured SO2 in the UTLS at background (non-volcanic) conditions with a precision of 2 ppt at 10 s and an overall uncertainty determined primarily by instrument drifts of +/- (16% + 0.9 ppt).

High incidence of rainbow glare after femtosecond laser assisted-LASIK using the upgraded FS200 femtosecond laser.

Science.gov (United States)

Zhang, Yu; Chen, Yue-Guo

2018-03-05

To compare the incidence of rainbow glare (RG) after femtosecond laser assisted-LASIK (FS-LASIK) using the upgraded FS200 femtosecond laser with different flap cut parameter settings. A consecutive series of 129 patients (255 eyes) who underwent FS-LASIK for correcting myopia and/or astigmatism using upgraded WaveLight FS200 femtosecond laser with the original settings was included in group A. Another consecutive series of 129 patients (255 eyes) who underwent FS-LASIK using upgraded WaveLight FS200 femtosecond laser with flap cut parameter settings changed (decreased pulse energy, spot and line separation) was included in group B. The incidence and fading time of RG, confocal microscopic image and postoperative clinical results were compared between the two groups. There were no differences between the two groups in age, baseline refraction, excimer laser ablation depth, postoperative uncorrected visual acuity and refraction. The incidence rate of RG in group A (35/255, 13.73%) was significantly higher than that in group B (4/255, 1.57%) (P  0.05).The confocal microscopic images showed wider laser spot spacing in group A than group B. The incidence of RG was significantly correlated with age and grouping (P laser with original flap cut parameter settings could increase the incidence of RG. The narrower grating size and lower pulse energy could ameliorate this side effect.

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.

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.

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.

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.

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

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

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

A Clinical and Confocal Microscopic Comparison of Transepithelial PRK and LASEK for Myopia

Directory of Open Access Journals (Sweden)

Safak Korkmaz

2014-01-01

Full Text Available Purpose. To compare the clinical and confocal microscopic results of transepithelial PRK versus LASEK for correction of myopia. Materials and Methods. Twelve patients with myopia received transepithelial PRK in one eye and LASEK in the other. In transepithelial PRK-treated eyes, the corneal epithelium was removed with 40 microns of excimer laser ablation and in LASEK-treated eyes with 25-second application of 18% ethanol. Time to epithelial healing, ocular discomfort, uncorrected and best corrected visual acuities, manifest refraction, haze, greyscale value, and keratocyte apoptosis in confocal microscopy were recorded. Results. The mean time to epithelial healing was significantly longer after LASEK (4.00 ± 0.43 versus 3.17 ± 0.6 days. On day 1, ocular discomfort was significantly higher after transepithelial PRK. The grade of haze, keratocyte apoptosis, and greyscale value in confocal microscopy were significantly higher in transepithelial PRK-treated eyes at 1 month. All transepithelial PRK- and LASEK-treated eyes achieved 20/25 or better UCVA and were within ±1.00 D of emmetropia at final visits. Conclusions. Both transepithelial PRK and LASEK offer effective correction of myopia at 1 year. However, LASEK appeared to induce less discomfort and less intense wound healing in the early postoperative period.

A Clinical and Confocal Microscopic Comparison of Transepithelial PRK and LASEK for Myopia.

Science.gov (United States)

Korkmaz, Safak; Bilgihan, Kamil; Sul, Sabahattin; Hondur, Ahmet

2014-01-01

Purpose. To compare the clinical and confocal microscopic results of transepithelial PRK versus LASEK for correction of myopia. Materials and Methods. Twelve patients with myopia received transepithelial PRK in one eye and LASEK in the other. In transepithelial PRK-treated eyes, the corneal epithelium was removed with 40 microns of excimer laser ablation and in LASEK-treated eyes with 25-second application of 18% ethanol. Time to epithelial healing, ocular discomfort, uncorrected and best corrected visual acuities, manifest refraction, haze, greyscale value, and keratocyte apoptosis in confocal microscopy were recorded. Results. The mean time to epithelial healing was significantly longer after LASEK (4.00 ± 0.43 versus 3.17 ± 0.6 days). On day 1, ocular discomfort was significantly higher after transepithelial PRK. The grade of haze, keratocyte apoptosis, and greyscale value in confocal microscopy were significantly higher in transepithelial PRK-treated eyes at 1 month. All transepithelial PRK- and LASEK-treated eyes achieved 20/25 or better UCVA and were within ±1.00 D of emmetropia at final visits. Conclusions. Both transepithelial PRK and LASEK offer effective correction of myopia at 1 year. However, LASEK appeared to induce less discomfort and less intense wound healing in the early postoperative period.

Compensation of inhomogeneous fluorescence signal distribution in 2D images acquired by confocal microscopy

Czech Academy of Sciences Publication Activity Database

Michálek, Jan; Čapek, Martin; Kubínová, Lucie

2011-01-01

Roč. 74, č. 9 (2011), s. 831-838 ISSN 1059-910X R&D Projects: GA ČR(CZ) GA102/08/0691; GA ČR(CZ) GA304/09/0733; GA MŠk(CZ) LC06063; GA MŠk(CZ) ME09010 Institutional research plan: CEZ:AV0Z50110509 Keywords : confocal laser scanning microscopy * image enhancement * morphology filters Subject RIV: JC - Computer Hardware ; Software Impact factor: 1.792, year: 2011

Visualization and quantitation of abundant macroautophagy in virus-infected cells by confocal three-dimensional fluorescence imaging.

Science.gov (United States)

Jackson, Wallen; Yamada, Masaki; Moninger, Thomas; Grose, Charles

2013-10-01

Varicella-zoster virus (VZV) is a human herpesvirus. Primary infection causes varicella (chickenpox), a viremic illness typified by an exanthem consisting of several hundred vesicles. When VZV reactivates from latency in the spinal ganglia during late adulthood, the emerging virus causes a vesicular dermatomal rash (herpes zoster or shingles). To expand investigations of autophagy during varicella and zoster, newer 3D imaging technology was combined with laser scanning confocal microscopy to provide animations of autophagosomes in the vesicular rash. First, the cells were immunolabeled with antibodies against VZV proteins and the LC3 protein, an integral autophagosomal protein. Antibody reagents lacking activity against the human blood group A1 antigen were selected. After laser excitation of the samples, optimized emission detection bandwidths were configured by Zeiss Zen control software. Confocal Z-stacks comprising up to 40 optical slices were reconstructed into 3D animations with the aid of Imaris software. With this imaging technology, individual autophagosomes were clearly detectable as spheres within each vesicular cell. To enumerate the number of autophagosomes, data sets from 50 cells were reconstructed as 3D fluorescence images and analyzed with MeasurementPro software. The mean number of autophagosomes per infected vesicular cell was >100, although over 200 autophagosomes were seen in a few cells. In summary, macroautophagy was easily quantitated within VZV-infected cells after immunolabeling and imaging by 3D confocal animation technology. These same 3D imaging techniques will be applicable for investigations of autophagy in other virus-infected cells. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

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

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.

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.

4Pi-confocal microscopy of live cells

Science.gov (United States)

Bahlmann, Karsten; Jakobs, Stefan; Hell, Stefan W.

2002-06-01

By coherently adding the spherical wavefronts of two opposing lenses, two-photon excitation 4Pi-confocal fluorescence microscopy has achieved three-dimensional imaging with an axial resolution 3-7 times better than confocal microscopy. So far this improvement was possible only in glycerol-mounted, fixed cells. Here we report 4Pi-confocal microscopy of watery objects and its application to the imaging of live cells. Water immersion 4Pi-confocal microscopy of membrane stained live Escherichia coli bacteria attains a 4.3 fold better axial resolution as compared to the best water immersion confocal microscope. The resolution enhancement results into a vastly improved three-dimensional representation of the bacteria. The first images of live biological samples with an all-directional resolution in the 190-280 nm range are presented here, thus establishing a new resolution benchmark in live cell microscopy.

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

Insights into esophagus tissue architecture using two-photon confocal microscopy

Science.gov (United States)

Liu, Nenrong; Wang, Yue; Feng, Shangyuan; Chen, Rong

2013-08-01

In this paper, microstructures of human esophageal mucosa were evaluated using the two-photon laser scanning confocal microscopy (TPLSCM), based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). The distribution of epithelial cells, muscle fibers of muscularis mucosae has been distinctly obtained. Furthermore, esophageal submucosa characteristics with cancer cells invading into were detected. The variation of collagen, elastin and cancer cells is very relevant to the pathology in esophagus, especially early esophageal cancer. Our experimental results indicate that the MPM technique has the much more advantages for label-free imaging, and has the potential application in vivo in the clinical diagnosis and monitoring of early esophageal cancer.

Microsphere imaging with confocal microscopy and two photon microscopy

International Nuclear Information System (INIS)

Chun, Hyung Su; An, Kyung Won; Lee, Jai Hyung

2002-01-01

We have acquired images of polystyrene and fused-silica microsphere by using conventional optical microscopy, confocal microscopy and two-photon microscopy, and performed comparative analysis of these images. Different from conventional optical microscopy, confocal and two-photon microscopy had good optical sectioning capability. In addition, confocal microscopy and two-photon microscopy had better lateral resolution than conventional optical microscopy. These results are attributed to confocality and nonlinearity of confocal microscopy and two photon microscopy, respectively.

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)

How to improve a critical performance for an ExoMars 2020 Scientific Instrument (RLS). Raman Laser Spectrometer Signal to Noise Ratio (SNR) Optimization

Science.gov (United States)

Canora, C. P.; Moral, A. G.; Rull, F.; Maurice, S.; Hutchinson, I.; Ramos, G.; López-Reyes, G.; Belenguer, T.; Canchal, R.; Prieto, J. A. R.; Rodriguez, P.; Santamaria, P.; Berrocal, A.; Colombo, M.; Gallago, P.; Seoane, L.; Quintana, C.; Ibarmia, S.; Zafra, J.; Saiz, J.; Santiago, A.; Marin, A.; Gordillo, C.; Escribano, D.; Sanz-Palominoa, M.

2017-09-01

The Raman Laser Spectrometer (RLS) is one of the Pasteur Payload instruments, within the ESA's Aurora Exploration Programme, ExoMars mission. Raman spectroscopy is based on the analysis of spectral fingerprints due to the inelastic scattering of light when interacting with matter. RLS is composed by Units: SPU (Spectrometer Unit), iOH (Internal Optical Head), and ICEU (Instrument Control and Excitation Unit) and the harnesses (EH and OH). The iOH focuses the excitation laser on the samples and collects the Raman emission from the sample via SPU (CCD) and the video data (analog) is received, digitalizing it and transmiting it to the processor module (ICEU). The main sources of noise arise from the sample, the background, and the instrument (Laser, CCD, focuss, acquisition parameters, operation control). In this last case the sources are mainly perturbations from the optics, dark signal and readout noise. Also flicker noise arising from laser emission fluctuations can be considered as instrument noise. In order to evaluate the SNR of a Raman instrument in a practical manner it is useful to perform end-to-end measurements on given standards samples. These measurements have to be compared with radiometric simulations using Raman efficiency values from literature and taking into account the different instrumental contributions to the SNR. The RLS EQM instrument performances results and its functionalities have been demonstrated in accordance with the science expectations. The Instrument obtained SNR performances in the RLS EQM will be compared experimentally and via analysis, with the Instrument Radiometric Model tool. The characterization process for SNR optimization is still on going. The operational parameters and RLS algorithms (fluorescence removal and acquisition parameters estimation) will be improved in future models (EQM-2) until FM Model delivery.

Morphological change study on root surfaces treated with curettes, sonic instruments or Er:YAG laser

International Nuclear Information System (INIS)

Guimaraes Filho, Arlindo Lopes

2004-01-01

Periodontal disease is caused by dental plaque and dental calculus on roots surfaces, specially on cervical areas. As dental plaque is the main cause and dental calculus a secondary one, it is practically impossible to separate one factor to the other one. In order to get periodontal tissue health it is necessary to eliminate dental plaque and calculus from root surfaces. In this sense, Er:YAG laser comes in as an excellent way to control periodontal disease, not only, by removing calculus and dental plaque but also for its bacteria reduction. The aim of this study is to compare, by S.E.M., root surfaces changing when they are treated with curettes and ultrasonic scaling or Er:YAG laser irradiation with two different energy levels of 60 mJ/pulse and 100 mJ/pulse and repetition tax of 10 Hz (in the display). It is also objective of this study to check a possible thermic damage to pulp tissue when the roots surfaces are irradiated with Er:YAG laser. We used for this study, five human dental roots, each one of them were cut into four samples, giving us a total of twenty samples, which were divided in five groups of four samples each one. The control group, we did not indicated any kind of treatment. The first group, the roots samples were scaled and planned with Gracey curettes 5/6 and 7/8. The second group, the roots samples were treated with ultrasonic instruments. The third group was irradiated with Er:YAG laser using 60 mJ/pulse , 10 Hz and energy density of 4 J/cm 2 (approximated). The fourth group was irradiated with Er:YAG laser using 100 mJ/pulse, 10 Hz and energy density of 7 J/cm 2 (approximated). The results analysis showed that roots scaling either with Gracey curettes or with ultrasonic instruments created smear layer covering roots surfaces; roots surfaces irradiated with Er:YAG laser showed few roughness in the third group; roots surfaces irradiated with Er:YAG laser showed no smear layer and the Er:YAG laser irradiation did not bring any thermic damage

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

Mars Science Laboratory Using Laser Instrument, Artist's Concept

Science.gov (United States)

2007-01-01

This artist's conception of NASA's Mars Science Laboratory portrays use of the rover's ChemCam instrument to identify the chemical composition of a rock sample on the surface of Mars. ChemCam is innovative for planetary exploration in using a technique referred to as laser breakdown spectroscopy to determine the chemical composition of samples from distances of up to about 8 meters (25 feet) away. ChemCam is led by a team at the Los Alamos National Laboratory and the Centre d'Etude Spatiale des Rayonnements in Toulouse, France. Mars Science Laboratory, a mobile robot for investigating Mars' past or present ability to sustain microbial life, is in development at NASA's Jet Propulsion Laboratory for a launch opportunity in 2009. The mission is managed by JPL, a division of the California Institute of Technology, Pasadena, Calif., for the NASA Science Mission Directorate, Washington.

The Fiber Optic System for the Advanced Topographic Laser Altimeter System (ATLAS) Instrument

Science.gov (United States)

Ott, Melanie N.; Thomes, Joe; Onuma, Eleanya; Switzer, Robert; Chuska, Richard; Blair, Diana; Frese, Erich; Matyseck, Marc

2016-01-01

The Advanced Topographic Laser Altimeter System (ATLAS) Instrument has been in integration and testing over the past 18 months in preparation for the Ice, Cloud and Land Elevation Satellite - 2 (ICESat-2) Mission, scheduled to launch in 2017. ICESat-2 is the follow on to ICESat which launched in 2003 and operated until 2009. ATLAS will measure the elevation of ice sheets, glaciers and sea ice or the "cryosphere" (as well as terrain) to provide data for assessing the earth's global climate changes. Where ICESat's instrument, the Geo-Science Laser Altimeter (GLAS) used a single beam measured with a 70 m spot on the ground and a distance between spots of 170 m, ATLAS will measure a spot size of 10 m with a spacing of 70 cm using six beams to measure terrain height changes as small as 4 mm. The ATLAS pulsed transmission system consists of two lasers operating at 532 nm with transmitter optics for beam steering, a diffractive optical element that splits the signal into 6 separate beams, receivers for start pulse detection and a wavelength tracking system. The optical receiver telescope system consists of optics that focus all six beams into optical fibers that feed a filter system that transmits the signal via fiber assemblies to the detectors. Also included on the instrument is a system that calibrates the alignment of the transmitted pulses to the receiver optics for precise signal capture. The larger electro optical subsystems for transmission, calibration, and signal receive, stay aligned and transmitting sufficiently due to the optical fiber system that links them together. The robust design of the fiber optic system, consisting of a variety of multi fiber arrays and simplex assemblies with multiple fiber core sizes and types, will enable the system to maintain consistent critical alignments for the entire life of the mission. Some of the development approaches used to meet the challenging optical system requirements for ATLAS are discussed here.

The fiber optic system for the Advanced Topographic Laser Altimeter System (ATLAS) instrument.

Science.gov (United States)

Ott, Melanie N; Thomes, Joe; Onuma, Eleanya; Switzer, Robert; Chuska, Richard; Blair, Diana; Frese, Erich; Matyseck, Marc

2016-08-28

The Advanced Topographic Laser Altimeter System (ATLAS) Instrument has been in integration and testing over the past 18 months in preparation for the Ice, Cloud and Land Elevation Satellite - 2 (ICESat-2) Mission, scheduled to launch in 2017. ICESat-2 is the follow on to ICESat which launched in 2003 and operated until 2009. ATLAS will measure the elevation of ice sheets, glaciers and sea ice or the "cryosphere" (as well as terrain) to provide data for assessing the earth's global climate changes. Where ICESat's instrument, the Geo-Science Laser Altimeter (GLAS) used a single beam measured with a 70 m spot on the ground and a distance between spots of 170 m, ATLAS will measure a spot size of 10 m with a spacing of 70 cm using six beams to measure terrain height changes as small as 4 mm.[1] The ATLAS pulsed transmission system consists of two lasers operating at 532 nm with transmitter optics for beam steering, a diffractive optical element that splits the signal into 6 separate beams, receivers for start pulse detection and a wavelength tracking system. The optical receiver telescope system consists of optics that focus all six beams into optical fibers that feed a filter system that transmits the signal via fiber assemblies to the detectors. Also included on the instrument is a system that calibrates the alignment of the transmitted pulses to the receiver optics for precise signal capture. The larger electro optical subsystems for transmission, calibration, and signal receive, stay aligned and transmitting sufficiently due to the optical fiber system that links them together. The robust design of the fiber optic system, consisting of a variety of multi fiber arrays and simplex assemblies with multiple fiber core sizes and types, will enable the system to maintain consistent critical alignments for the entire life of the mission. Some of the development approaches used to meet the challenging optical system requirements for ATLAS are discussed here.

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.

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.

Microscopia confocal en operados de queratoplastia perforante Confocal microscopy in patients operated from penetrating keratoplasty

Directory of Open Access Journals (Sweden)

Zulema Gómez Castillo

2009-06-01

Full Text Available La microscopia confocal es un examen exploratorio, práctico y poco invasivo que permite conocer las características microscópicas del tejido corneal después del trasplante, por lo que constituye una herramienta muy útil en el manejo de los pacientes operados de queratoplastia. El presente trabajo tiene como finalidad describir las características del tejido corneal en pacientes operados de este tipo de trasplante, mediante la microscopia confocal in vivo. MÉTODOS: Se realizó un estudio descriptivo, de corte transversal, en 40 ojos de 40 pacientes operados de queratoplastia perforante, en el Servicio de Córnea del Instituto Cubano de Oftalmología "Ramón Pando Ferrer", de marzo de 2006 a marzo de 2007. Se confeccionó una historia clínica oftalmológica y se les realizó a todos el examen de microscopia confocal en el injerto corneal con el microscopio confocal CONFOSCAN 4. RESULTADOS: La queratopatía bullosa pseudofáquica fue la afección más frecuente previa a la cirugía y estuvo presente en el 77,5 % de los pacientes. En el 72,5 % de los intervenidos se encontró una disminución del grosor corneal. El epitelio presentó alteraciones en el 62,5 % de los pacientes. Todos presentaron afectación de la forma y el tamaño celular endotelial. En el 82,5 % de los pacientes se observó ausencia de plexos nerviosos. CONCLUSIONES: La microscopia confocal como nueva ciencia en el campo de la oftalmología, favorece el seguimiento evolutivo de las queratoplastias perforantes y con esto no solo a prevenir la aparición de posibles complicaciones, sino además de garantizar el éxito de la cirugía y la función refractiva de la córnea.Confocal microscopy is a practical, exploratory and less invassive examination that allows finding out the microscopic characteristics of the corneal tissue after transplantation, so it is a very useful tool for the management of patients operated from keratoplasty. The present paper was aimed at describing

Application of function-oriented roughness parameters using confocal microscopy

Directory of Open Access Journals (Sweden)

K. Klauer

2018-06-01

Full Text Available Optical measuring instruments are widely used for the functional characterization of surface topography. However, due to the interaction of the surface with the incident light, effects occur that can influence the measured topography height values and the obtained surface texture parameters. Therefore, we describe a systematic investigation of the influences of optical surface topography measurement on the acquisition of function-oriented roughness parameters. The same evaluation areas of varying cylinder liners which represent a typical application of function-oriented roughness parameters were measured with a confocal microscope and a stylus instrument. Functional surface texture parameters as given in the standards ISO 13565–2, ISO 13565–3 and ISO 25178–2 were evaluated for both measurement methods and compared. The transmission of specific surface features was described and a correlation analysis for the surface topographies obtained with the different measurement methods and their resulting functional roughness parameters was carried out. Keywords: Functional surface characterization, Optical metrology, Topography measurement, Roughness

The Signal Detection and Control Circuit Design for Confocal Auto-Focus System

OpenAIRE

Yin Liu; Jin Yu; Zeqiang Mo

2016-01-01

Based on the demands of Confocal Auto-Focus system, the implementation method of signal measurement circuit and control circuit is given. Using the high performance instrumental amplifier AD620BN, low noise precision FET Op amplifier AD795JRZ and ultralow offset voltage Op amplifier OP07EP, a signal measurement circuit used to converse the two differential light intensity signal to electric signal is designed. And a control circuit which takes MCU MSP430F149 as core processes the former signa...

Automated Microscopy: Macro Language Controlling a Confocal Microscope and its External Illumination: Adaptation for Photosynthetic Organisms

Czech Academy of Sciences Publication Activity Database

Steinbach, Gabor; Kaňa, Radek

2016-01-01

Roč. 22, č. 2 (2016), s. 258-263 ISSN 1431-9276 R&D Projects: GA ČR GAP501/12/0304; GA MŠk EE2.3.30.0059; GA MŠk ED2.1.00/03.0110; GA MŠk(CZ) LO1416 Institutional support: RVO:61388971 Keywords : automated microscopy * remote controlled microscopy * confocal microscopy Subject RIV: BH - Optics, Masers, Lasers Impact factor: 1.891, year: 2016

Combining total internal reflection sum frequency spectroscopy spectral imaging and confocal fluorescence microscopy.

Science.gov (United States)

Allgeyer, Edward S; Sterling, Sarah M; Gunewardene, Mudalige S; Hess, Samuel T; Neivandt, David J; Mason, Michael D

2015-01-27

Understanding surface and interfacial lateral organization in material and biological systems is critical in nearly every field of science. The continued development of tools and techniques viable for elucidation of interfacial and surface information is therefore necessary to address new questions and further current investigations. Sum frequency spectroscopy (SFS) is a label-free, nonlinear optical technique with inherent surface specificity that can yield critical organizational information on interfacial species. Unfortunately, SFS provides no spatial information on a surface; small scale heterogeneities that may exist are averaged over the large areas typically probed. Over the past decade, this has begun to be addressed with the advent of SFS microscopy. Here we detail the construction and function of a total internal reflection (TIR) SFS spectral and confocal fluorescence imaging microscope directly amenable to surface investigations. This instrument combines, for the first time, sample scanning TIR-SFS imaging with confocal fluorescence microscopy.

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.

Optical feedback effects on terahertz quantum cascade lasers: modelling and applications

Science.gov (United States)

Rakić, Aleksandar D.; Lim, Yah Leng; Taimre, Thomas; Agnew, Gary; Qi, Xiaoqiong; Bertling, Karl; Han, She; Wilson, Stephen J.; Kundu, Iman; Grier, Andrew; Ikonić, Zoran; Valavanis, Alexander; Demić, Aleksandar; Keeley, James; Li, Lianhe H.; Linfield, Edmund H.; Davies, A. Giles; Harrison, Paul; Ferguson, Blake; Walker, Graeme; Prow, Tarl; Indjin, Dragan; Soyer, H. Peter

2016-11-01

Terahertz (THz) quantum cascade lasers (QCLs) are compact sources of radiation in the 1-5 THz range with significant potential for applications in sensing and imaging. Laser feedback interferometry (LFI) with THz QCLs is a technique utilizing the sensitivity of the QCL to the radiation reflected back into the laser cavity from an external target. We will discuss modelling techniques and explore the applications of LFI in biological tissue imaging and will show that the confocal nature of the QCL in LFI systems, with their innate capacity for depth sectioning, makes them suitable for skin diagnostics with the well-known advantages of more conventional confocal microscopes. A demonstration of discrimination of neoplasia from healthy tissue using a THz, LFI-based system in the context of melanoma is presented using a transgenic mouse model.

Development of the Potassium-Argon Laser Experiment (KArLE) Instrument for In Situ Geochronology

Science.gov (United States)

Cohen, Barbara A.; Li, Z.-H.; Miller, J. S.; Brinckerhoff, W. B.; Clegg, S. M.; Mahaffy, P. R.; Swindle, T. D.; Wiens, R. C.

2012-01-01

Absolute dating of planetary samples is an essential tool to establish the chronology of geological events, including crystallization history, magmatic evolution, and alteration. Traditionally, geochronology has only been accomplishable on samples from dedicated sample return missions or meteorites. The capability for in situ geochronology is highly desired, because it will allow one-way planetary missions to perform dating of large numbers of samples. The success of an in situ geochronology package will not only yield data on absolute ages, but can also complement sample return missions by identifying the most interesting rocks to cache and/or return to Earth. In situ dating instruments have been proposed, but none have yet reached TRL 6 because the required high-resolution isotopic measurements are very challenging. Our team is now addressing this challenge by developing the Potassium (K) - Argon Laser Experiment (KArLE) under the NASA Planetary Instrument Definition and Development Program (PIDDP), building on previous work to develop a K-Ar in situ instrument [1]. KArLE uses a combination of several flight-proven components that enable accurate K-Ar isochron dating of planetary rocks. KArLE will ablate a rock sample, determine the K in the plasma state using laser-induced breakdown spectroscopy (LIBS), measure the liberated Ar using quadrupole mass spectrometry (QMS), and relate the two by the volume of the ablated pit using an optical method such as a vertical scanning interferometer (VSI). Our preliminary work indicates that the KArLE instrument will be capable of determining the age of several kinds of planetary samples to +/-100 Myr, sufficient to address a wide range of geochronology problems in planetary science.

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.

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.

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.

Confocal microlaparoscope for imaging the fallopian tube

Science.gov (United States)

Wu, Tzu-Yu; Rouse, Andrew R.; Chambers, Setsuko K.; Hatch, Kenneth D.; Gmitro, Arthur F.

2014-11-01

Recent evidence suggests that ovarian cancer can originate in the fallopian tube. Unlike many other cancers, poor access to the ovary and fallopian tubes has limited the ability to study the progression of this deadly disease and to diagnosis it during the early stage when it is most amenable to therapy. A rigid confocal microlaparoscope system designed to image the epithelial surface of the ovary in vivo was previously reported. A new confocal microlaparoscope with an articulating distal tip has been developed to enable in vivo access to human fallopian tubes. The new microlaparoscope is compatible with 5-mm trocars and includes a 2.2-mm-diameter articulating distal tip consisting of a bare fiber bundle and an automated dye delivery system for fluorescence confocal imaging. This small articulating device should enable the confocal microlaparoscope to image early stage ovarian cancer arising inside the fallopian tube. Ex vivo images of animal tissue and human fallopian tube using the new articulating device are presented along with in vivo imaging results using the rigid confocal microlaparoscope system.

High harmonic terahertz confocal gyrotron with nonuniform electron beam

Energy Technology Data Exchange (ETDEWEB)

Fu, Wenjie; Guan, Xiaotong; Yan, Yang [THz Research Center, School of Physical Electronics, University of Electronic Science and Technology of China, Chengdu 610054 (China)

2016-01-15

The harmonic confocal gyrotron with nonuniform electron beam is proposed in this paper in order to develop compact and high power terahertz radiation source. A 0.56 THz third harmonic confocal gyrotron with a dual arc section nonuniform electron beam has been designed and investigated. The studies show that confocal cavity has extremely low mode density, and has great advantage to operate at high harmonic. Nonuniform electron beam is an approach to improve output power and interaction efficiency of confocal gyrotron. A dual arc beam magnetron injection gun for designed confocal gyrotron has been developed and presented in this paper.

A new instrument of VUV laser desorption/ionization mass spectrometry imaging with micrometer spatial resolution and low level of molecular fragmentation.

Science.gov (United States)

Wang, Jia; Liu, Feng; Mo, Yuxiang; Wang, Zhaoying; Zhang, Sichun; Zhang, Xinrong

2017-11-01

Mass spectrometry imaging (MSI) has important applications in material research, biology, and medicine. The MSI method based on UV laser desorption/ionization (UVLDI) can obtain images of intact samples, but has a high level of molecular fragmentation. In this work, we report a new MSI instrument that uses a VUV laser (125.3 nm) as a desorption/ionization source to exploit its advantages of high single photon energy and small focus size. The new instrument was tested by the mass spectra of Nile red and FGB (Fibrinogen beta chain) samples and mass spectrometric images of a fly brain section. For the tested samples, the VUVDI method offers lower levels of molecular fragmentations and higher sensitivities than those of the UVLDI method and second ion mass spectrometry imaging method using a Bi 3 + beam. The ablation crater produced by the focused VUV laser on a quartz plate has an area of 10 μm 2 . The VUV laser is prepared based on the four-wave mixing method using three collimated laser beams and a heated Hg cell.

A new instrument of VUV laser desorption/ionization mass spectrometry imaging with micrometer spatial resolution and low level of molecular fragmentation

Science.gov (United States)

Wang, Jia; Liu, Feng; Mo, Yuxiang; Wang, Zhaoying; Zhang, Sichun; Zhang, Xinrong

2017-11-01

Mass spectrometry imaging (MSI) has important applications in material research, biology, and medicine. The MSI method based on UV laser desorption/ionization (UVLDI) can obtain images of intact samples, but has a high level of molecular fragmentation. In this work, we report a new MSI instrument that uses a VUV laser (125.3 nm) as a desorption/ionization source to exploit its advantages of high single photon energy and small focus size. The new instrument was tested by the mass spectra of Nile red and FGB (Fibrinogen beta chain) samples and mass spectrometric images of a fly brain section. For the tested samples, the VUVDI method offers lower levels of molecular fragmentations and higher sensitivities than those of the UVLDI method and second ion mass spectrometry imaging method using a Bi3+ beam. The ablation crater produced by the focused VUV laser on a quartz plate has an area of 10 μm2. The VUV laser is prepared based on the four-wave mixing method using three collimated laser beams and a heated Hg cell.

The Development and Deployment of a Ground-Based, Laser-Induced Fluorescence Instrument for the In Situ Detection of Iodine Monoxide Radicals

Science.gov (United States)

Thurlow, M. E.; Co, D. T.; O'Brien, A. S.; Hannun, R. A.; Lapson, L. B.; Hanisco, T. F.; Anderson, J. G.

2014-01-01

High abundances of iodine monoxide (IO) are known to exist and to participate in local photochemistry of the marine boundary layer. Of particular interest are the roles IO plays in the formation of new particles in coastal marine environments and in depletion episodes of ozone and mercury in the Arctic polar spring. This paper describes a ground-based instrument that measures IO at mixing ratios less than one part in 1012. The IO radical is measured by detecting laser-induced fluorescence at wavelengths longer that 500 nm. Tunable visible light is used to pump the A23/2 (v = 2) ? X23/2 (v = 0) transition of IO near 445 nm. The laser light is produced by a solid-state, Nd:YAG-pumped Ti:Sapphire laser at 5 kHz repetition rate. The laser-induced fluorescence instrument performs reliably with very high signal-to-noise ratios (>10) achieved in short integration times (<1 min). The observations from a validation deployment to the Shoals Marine Lab on Appledore Island, ME are presented and are broadly consistent with in situ observations from European Coastal Sites. Mixing ratios ranged from the instrumental detection limit (<1 pptv) to 10 pptv. These data represent the first in situ point measurements of IO in North America.

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.

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.

Green-synthetized silver nanoparticles for Nanoparticle-Enhanced Laser Induced Breakdown Spectroscopy (NELIBS) using a mobile instrument

Science.gov (United States)

Poggialini, F.; Campanella, B.; Giannarelli, S.; Grifoni, E.; Legnaioli, S.; Lorenzetti, G.; Pagnotta, S.; Safi, A.; Palleschi, V.

2018-03-01

When compared to other analytical techniques, LIBS shows relatively low precision and, generally, high Limits of Detection (LODs). Until recently, the attempts in improving the LIBS performances have been based on the use of more stable/powerful lasers, high sensitivity detectors or controlled environmental parameters. This can hinder the competitiveness of LIBS by increasing the instrumental setup cost and the difficulty of operation. Sample treatment has proved to be a viable and simple way to increase the LIBS signal; in particular, the Nanoparticle-Enhanced Laser Induced Breakdown Spectroscopy (NELIBS) methodology uses a deposition of metal nanoparticles on the sample to greatly increase the emission of the LIBS plasma. In this work, we used a simple, fast, "green" and low-cost method to synthetize silver nanoparticles by using coffee extract as reducing agents for a silver nitrate solution. This allowed us to obtain nanoparticles of about 25 nm in diameter. We then explored the application of such nanoparticles to the NELIBS analysis of metallic samples with a mobile LIBS instrument. By adjusting the laser parameters and optimizing the sample preparation procedure, we obtained a NELIBS signal that is 4 times the LIBS one. This showed the potential of green-synthetized nanoparticle for NELIBS applications and suggests the possibility of an in-situ application of the technique.

SU-F-T-665: Confocal Microscopy Imaging of Cell Cycle Distribution in Cells Treated with Pegylated Gold Nanoshells

Energy Technology Data Exchange (ETDEWEB)

Sadetaporn, D [Rice University, Houston, TX (United States); The University of Texas MD Anderson Cancer Center, Houston, TX (United States); Flint, D; McFadden, C; Sawakuchi, G [The University of Texas MD Anderson Cancer Center, Houston, TX (United States); Asaithamby, A [UT Southwestern Medical Center, Dallas, TX (United States)

2016-06-15

Purpose: To use confocal microscopy to distinguish cells in different phases of the cell cycle before and after treatment with pegylated gold nanoshells (PEG-AuNSs). Methods: Transfected fibrosarcoma cells (HT1080-EYFP-53BP1-FUCCI) were cultured in T-25 flasks and seeded in glass bottom dishes. These cells express the fluorescent probe AmCyan during the G2/S phases of the cell cycle, mCherry during the G1 phase, and EYFP tagged to the DNA repair protein 53BP1. After allowing cells 4 h to adhere to dishes, PEG-AuNS (Nanospectra Biosciences, Houston, TX) at a concentration of 0.15 OD were administered. At time points of 8, 16 and 24 h following treatment, the PEG-AuNS-treated and control samples were washed with phosphate buffered saline (PBS) and fixed using 4% paraformaldehyde in PBS. Samples were imaged with an Olympus FV1200 confocal microscope using 473, 543, and 641 nm excitation lasers. We used band-pass filters to select AmCyan and mCherry fluorescence. Reflection from the 641 nm laser was used to detect PEG-AuNSs. Z-stack images were analyzed to assess cell cycle distribution through fluorescent probe expression. Live cells were imaged after PEG-AuNS treatment using a confocal microscope with a stage top CO2 incubator. Results: We were able to obtain high-resolution images of cells with internalized AuNSs. We were also able to distinguish cells in different phases of the cell cycle. Conclusion: This work demonstrates a new assay to investigate the effect of AuNSs on the cell cycle phase in live cells. Future work will employ confocal microscopy and flow cytometry to focus on effects of AuNS treatment on cell cycle distribution. This research was supported by the Sister Institution Network Fund and the Center for Radiation Oncology Research at The University of Texas MD Anderson Cancer Center and Cancer Prevention and Research Institute of Texas. Gabriel Sawakuchi has research support from Elekta Inc.

SU-F-T-665: Confocal Microscopy Imaging of Cell Cycle Distribution in Cells Treated with Pegylated Gold Nanoshells

International Nuclear Information System (INIS)

Sadetaporn, D; Flint, D; McFadden, C; Sawakuchi, G; Asaithamby, A

2016-01-01

Purpose: To use confocal microscopy to distinguish cells in different phases of the cell cycle before and after treatment with pegylated gold nanoshells (PEG-AuNSs). Methods: Transfected fibrosarcoma cells (HT1080-EYFP-53BP1-FUCCI) were cultured in T-25 flasks and seeded in glass bottom dishes. These cells express the fluorescent probe AmCyan during the G2/S phases of the cell cycle, mCherry during the G1 phase, and EYFP tagged to the DNA repair protein 53BP1. After allowing cells 4 h to adhere to dishes, PEG-AuNS (Nanospectra Biosciences, Houston, TX) at a concentration of 0.15 OD were administered. At time points of 8, 16 and 24 h following treatment, the PEG-AuNS-treated and control samples were washed with phosphate buffered saline (PBS) and fixed using 4% paraformaldehyde in PBS. Samples were imaged with an Olympus FV1200 confocal microscope using 473, 543, and 641 nm excitation lasers. We used band-pass filters to select AmCyan and mCherry fluorescence. Reflection from the 641 nm laser was used to detect PEG-AuNSs. Z-stack images were analyzed to assess cell cycle distribution through fluorescent probe expression. Live cells were imaged after PEG-AuNS treatment using a confocal microscope with a stage top CO2 incubator. Results: We were able to obtain high-resolution images of cells with internalized AuNSs. We were also able to distinguish cells in different phases of the cell cycle. Conclusion: This work demonstrates a new assay to investigate the effect of AuNSs on the cell cycle phase in live cells. Future work will employ confocal microscopy and flow cytometry to focus on effects of AuNS treatment on cell cycle distribution. This research was supported by the Sister Institution Network Fund and the Center for Radiation Oncology Research at The University of Texas MD Anderson Cancer Center and Cancer Prevention and Research Institute of Texas. Gabriel Sawakuchi has research support from Elekta Inc.

Confocal filtering in cathodoluminescence microscopy of nanostructures

Science.gov (United States)

Narváez, Angela C.; Weppelman, I. Gerward C.; Moerland, Robert J.; Hoogenboom, Jacob P.; Kruit, Pieter

2014-06-01

Cathodoluminescence (CL) microscopy allows optical characterization of nanostructures at high spatial resolution. At the nanoscale, a main challenge of the technique is related to the background CL generated within the sample substrate. Here, we implement confocal detection of the CL signal to minimize the background contribution to the measurement. Nano-phosphors were used as point sources to evaluate the filtering capabilities of our confocal CL system, obtaining an axial intensity profile with 2.7 μm full width at half maximum for the central peak, in good correspondence with theoretical expectations. Considering the electron interaction volume, we found that the confocal filter becomes effective for electron energies above 20 keV, when using a 25 μm pinhole (0.86 Airy units). To illustrate our approach, we present confocal CL imaging of gold nanowires and triangular shaped plates deposited on an indium-tin oxide covered glass substrate, comparing the images with those obtained in standard unfiltered CL detection. The results show that confocal CL microscopy is a valuable tool for the investigation of nanostructures on highly cathodoluminescent substrates, widely used in biological and optical applications.

Confocal filtering in cathodoluminescence microscopy of nanostructures

Energy Technology Data Exchange (ETDEWEB)

Narváez, Angela C., E-mail: a.c.narvaez@tudelft.nl, E-mail: j.p.hoogenboom@tudelft.nl; Weppelman, I. Gerward C.; Moerland, Robert J.; Hoogenboom, Jacob P., E-mail: a.c.narvaez@tudelft.nl, E-mail: j.p.hoogenboom@tudelft.nl; Kruit, Pieter [Imaging Physics, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628CJ Delft (Netherlands)

2014-06-23

Cathodoluminescence (CL) microscopy allows optical characterization of nanostructures at high spatial resolution. At the nanoscale, a main challenge of the technique is related to the background CL generated within the sample substrate. Here, we implement confocal detection of the CL signal to minimize the background contribution to the measurement. Nano-phosphors were used as point sources to evaluate the filtering capabilities of our confocal CL system, obtaining an axial intensity profile with 2.7 μm full width at half maximum for the central peak, in good correspondence with theoretical expectations. Considering the electron interaction volume, we found that the confocal filter becomes effective for electron energies above 20 keV, when using a 25 μm pinhole (0.86 Airy units). To illustrate our approach, we present confocal CL imaging of gold nanowires and triangular shaped plates deposited on an indium-tin oxide covered glass substrate, comparing the images with those obtained in standard unfiltered CL detection. The results show that confocal CL microscopy is a valuable tool for the investigation of nanostructures on highly cathodoluminescent substrates, widely used in biological and optical applications.

Confocal endomicroscopy for in vivo microscopic analysis of upper gastrointestinal tract premalignant and malignant lesions.

Science.gov (United States)

Gheorghe, Cristian; Iacob, Razvan; Becheanu, Gabriel; Dumbrav Abreve, Mona

2008-03-01

Confocal LASER endomicroscopy (CLE) is a new endoscopic technique which allows subsurface in vivo microscopic analysis during ongoing endoscopy, using systemically or topically administered fluorescent agents. It allows targeted biopsies to be taken, potentially improving the diagnostic rate in certain gastrointestinal diseases. Worldwide experience with CLE for upper gastrointestinal malignant and premalignant lesions is still reduced. Potential clinical applications are presented, including diagnosis of NERD, Barrett's esophagus, atrophic gatritis, gastric intestinal metaplasia and dysplasia, gastric adenomatous or hyperplastic polyps, gastric cancer.

Image-guided, Laser-based Fabrication of Vascular-derived Microfluidic Networks

OpenAIRE

Heintz, Keely A.; Mayerich, David; Slater, John H.

2017-01-01

This detailed protocol outlines the implementation of image-guided, laser-based hydrogel degradation for the fabrication of vascular-derived microfluidic networks embedded in PEGDA hydrogels. Here, we describe the creation of virtual masks that allow for image-guided laser control; the photopolymerization of a micromolded PEGDA hydrogel, suitable for microfluidic network fabrication and pressure head-driven flow; the setup and use of a commercially available laser scanning confocal microscope...

Digital differential confocal microscopy based on spatial shift transformation.

Science.gov (United States)

Liu, J; Wang, Y; Liu, C; Wilson, T; Wang, H; Tan, J

2014-11-01

Differential confocal microscopy is a particularly powerful surface profilometry technique in industrial metrology due to its high axial sensitivity and insensitivity to noise. However, the practical implementation of the technique requires the accurate positioning of point detectors in three-dimensions. We describe a simple alternative based on spatial transformation of a through-focus series of images obtained from a homemade beam scanning confocal microscope. This digital differential confocal microscopy approach is described and compared with the traditional Differential confocal microscopy approach. The ease of use of the digital differential confocal microscopy system is illustrated by performing measurements on a 3D standard specimen. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

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.

Advanced Laser Architecture for Two-Step Laser Tandem Mass Spectrometer

Science.gov (United States)

Fahey, Molly E.; Li, Steven X.; Yu, Anthony W.; Getty, Stephanie A.

2016-01-01

Future astrobiology missions will focus on planets with significant astrochemical or potential astrobiological features, such as small, primitive bodies and the icy moons of the outer planets that may host diverse organic compounds. These missions require advanced instrument techniques to fully and unambiguously characterize the composition of surface and dust materials. Laser desorptionionization mass spectrometry (LDMS) is an emerging instrument technology for in situ mass analysis of non-volatile sample composition. A recent Goddard LDMS advancement is the two-step laser tandem mass spectrometer (L2MS) instrument to address the need for future flight instrumentation to deconvolve complex organic signatures. The L2MS prototype uses a resonance enhanced multi-photon laser ionization mechanism to selectively detect aromatic species from a more complex sample. By neglecting the aliphatic and inorganic mineral signatures in the two-step mass spectrum, the L2MS approach can provide both mass assignments and clues to structural information for an in situ investigation of non-volatile sample composition. In this paper we will describe our development effort on a new laser architecture that is based on the previously flown Lunar Orbiter Laser Altimeter (LOLA) laser transmitter for the L2MS instrument. The laser provides two discrete midinfrared wavelengths (2.8 m and 3.4 m) using monolithic optical parametric oscillators and ultraviolet (UV) wavelength (266 nm) on a single laser bench with a straightforward development path toward flight readiness.

Mode-mismatched confocal thermal-lens microscope with collimated probe beam

Energy Technology Data Exchange (ETDEWEB)

Cabrera, Humberto, E-mail: hcabrera@ictp.it [SPIE-ICTP Anchor Research Laboratory, International Centre for Theoretical Physics (ICTP), Strada Costiera 11, Trieste (Italy); Centro Multidisciplinartio de Ciencias, Instituto Venezolano de Investigaciones Científicas (IVIC), Mérida 5101 (Venezuela, Bolivarian Republic of); Korte, Dorota; Franko, Mladen [Laboratory for Environmental Research, University of Nova Gorica, Vipavska 13, 5000 Nova Gorica (Slovenia)

2015-05-15

We report a thermal lens microscope (TLM) based on an optimized mode-mismatched configuration. It takes advantage of the coaxial counter propagating tightly focused excitation and collimated probe beams, instead of both focused at the sample, as it is in currently known TLM setups. A simple mathematical model that takes into account the main features of the instrument is presented. The confocal detection scheme and the introduction of highly collimated probe beam allow enhancing the versatility, limit of detection (LOD), and sensitivity of the instrument. The theory is experimentally verified measuring ethanol’s absorption coefficient at 532.8 nm. Additionally, the presented technique is applied for detection of ultra-trace amounts of Cr(III) in liquid solution. The achieved LOD is 1.3 ppb, which represents 20-fold enhancement compared to transmission mode spectrometric techniques and a 7.5-fold improvement compared to previously reported methods for Cr(III) based on thermal lens effect.

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

EVIDÊNCIAS CIENTIFICAS SOBRE O USO DA ESPECTROSCOPIA RAMAN CONFOCAL IN VIVO NA PELE HUMANA

Directory of Open Access Journals (Sweden)

Aline Campos Pereira

2017-04-01

Full Text Available A Espectroscopia Raman Confocal (ERC é uma técnica totalmente não invasiva, eficaz na caracterização em tempo real dos arranjos químicos dos tecidos biológicos vivos. Com isso, o objetivo desse trabalho é destacar as pesquisas com uso da ERC. Foram selecionados e analisados das bases de dados: PubMed e Web of Science: 18 artigos científicos. Foram apresentados em dois quadros, obedecendo a ordem: nome dos autores, ano, revista, número de participantes, região espectral, tipo de sistema Raman Confocal, tipo e potência dos lasers. Todos os artigos reportados neste trabalham ressaltam que a ERC trata se de uma ferramenta valiosa, a qual fornece dados confiáveis. Conclui-se que existem poucos estudos científicos utilizando a ERC na pele humana, principalmente in vivo, apesar de fornecer informações em diferentes profundidades e obter dados com uma metodologia totalmente invasiva.

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

Self-Raman Nd:YVO4 Laser and Electro-Optic Technology for Space-Based Sodium Lidar Instrument

Science.gov (United States)

Krainak, Michael A.; Yu, Anthony W.; Janches, Diego; Jones, Sarah L.; Blagojevic, Branimir; Chen, Jeffrey

2014-01-01

We are developing a laser and electro-optic technology to remotely measure Sodium (Na) by adapting existing lidar technology with space flight heritage. The developed instrumentation will serve as the core for the planning of an Heliophysics mission targeted to study the composition and dynamics of Earth's mesosphere based on a spaceborne lidar that will measure the mesospheric Na layer. We present performance results from our diode-pumped tunable Q-switched self-Raman c-cut Nd:YVO4 laser with intra-cavity frequency doubling that produces multi-watt 589 nm wavelength output. The c-cut Nd:YVO4 laser has a fundamental wavelength that is tunable from 1063-1067 nanometers. A CW (Continuous Wave) External Cavity diode laser is used as a injection seeder to provide single-frequency grating tunable output around 1066 nanometers. The injection-seeded self-Raman shifted Nd:VO4 laser is tuned across the sodium vapor D2 line at 589 nanometers. We will review technologies that provide strong leverage for the sodium lidar laser system with strong heritage from the Ice Cloud and Land Elevation Satellite-2 (ICESat-2) Advanced Topographic Laser Altimeter System (ATLAS). These include a space-qualified frequency-doubled 9 watts-at-532-nanometer wavelength Nd:YVO4 laser, a tandem interference filter temperature-stabilized fused-silica-etalon receiver and high-bandwidth photon-counting detectors.

Confocal imaging of protein distributions in porous silicon optical structures

International Nuclear Information System (INIS)

De Stefano, Luca; D'Auria, Sabato

2007-01-01

The performances of porous silicon optical biosensors depend strongly on the arrangement of the biological probes into their sponge-like structures: it is well known that in this case the sensing species do not fill the pores but instead cover their internal surface. In this paper, the direct imaging of labelled proteins into different porous silicon structures by using a confocal laser microscope is reported. The distribution of the biological matter in the nanostructured material follows a Gaussian behaviour which is typical of the diffusion process in the porous media but with substantial differences between a porous silicon monolayer and a multilayer such as a Bragg mirror. Even if semi-quantitative, the results can be very useful in the design of the porous silicon based biosensing devices

Estudo comparativo histológico na prega vocal após incisão com instrumental a frio e com laser de CO2 em modelo animal Comparative histology study of the vocal folds after incision with cold instruments and CO2 laser in an animal model

Directory of Open Access Journals (Sweden)

Fernando C. C. Santos

2003-12-01

Full Text Available Dentre os vários tipos de laser usados em Medicina, o laser de CO2 é o mais usado na Otorrinolaringologia e Cirurgia de Cabeça e Pescoço. As vantagens de seu uso são a diminuição do sangramento, a diminuição do edema no pós-operatório e a facilidade de acesso ao campo operatório, entre outras. Desde os trabalhos de Jako e Strong em 1972(1,2, quando o laser de CO2 passou a ser usado no tratamento de papilomatose laríngea e de lesões malignas glóticas iniciais, suas indicações têm aumetado, principalmente em lesões benignas, a partir da alta tecnologia desenvolvida dos últimos anos como, por exemplo, a diminuição do microspot e o uso do superpulso, reduzindo conseqüentemente seu efeito térmico sobre os tecidos. MÉTODOS: Neste trabalho foram realizadas incisões com instrumental a frio e com laser de CO2 1 watt de modo contínuo e superpulso, em pregas vocais caninas e observado, através de cortes histológicos corados pelo método de Sirius Red, a quantidade de colágeno depositada sobre as mesmas. RESULTADOS: A quantidade de colágeno das pregas vocais foi maior do que no grupo controle, e estatisticamente maior no grupo de animais submetidos a procedimentos com instrumental a frio do que com laser de CO2. Não houve diferença estatística entre o grupo controle e o grupo submetido a incisões com instrumentos a frio. CONCLUSÃO: A microcirurgia de laringe com o laser de CO2, quando este é usado em baixa potência, com pequeno "microspot" e com superpulso, é um método seguro em relação à deposição de colágeno, quando comparado com instrumentos com lâmina a frio, obedecendo os princípios da fonomicrocirurgia.Among the several types of lasers employed in medicine today, the CO2 is the most widely used in otorhinolaryngology and head and neck surgeries. The advantages of its use are: the reduction in bleeding, reduction in post-operative edema and the easy access to the operation site, among others. Ever since

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...

The development and deployment of a ground-based, laser-induced fluorescence instrument for the in situ detection of iodine monoxide radicals

Energy Technology Data Exchange (ETDEWEB)

Thurlow, M. E., E-mail: thurlow@huarp.harvard.edu; Hannun, R. A.; Lapson, L. B.; Anderson, J. G. [Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138 (United States); Co, D. T. [Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138 (United States); Argonne-Northwestern Solar Energy Research Center and Department of Chemistry, Northwestern University, 2145 Sheridan Road, Evanston, Illinois 60208-3113 (United States); O' Brien, A. S. [Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138 (United States); Department of Civil and Environmental Engineering, Princeton University, Princeton, New Jersey 08544 (United States); Hanisco, T. F. [Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138 (United States); NASA Goddard Space Flight Center, Code 614, Greenbelt, Maryland 20771 (United States)

2014-04-15

High abundances of iodine monoxide (IO) are known to exist and to participate in local photochemistry of the marine boundary layer. Of particular interest are the roles IO plays in the formation of new particles in coastal marine environments and in depletion episodes of ozone and mercury in the Arctic polar spring. This paper describes a ground-based instrument that measures IO at mixing ratios less than one part in 10{sup 12}. The IO radical is measured by detecting laser-induced fluorescence at wavelengths longer that 500 nm. Tunable visible light is used to pump the A{sup 2}Π{sub 3/2} (v{sup ′} = 2) ← X{sup 2}Π{sub 3/2} (v{sup ″} = 0) transition of IO near 445 nm. The laser light is produced by a solid-state, Nd:YAG-pumped Ti:Sapphire laser at 5 kHz repetition rate. The laser-induced fluorescence instrument performs reliably with very high signal-to-noise ratios (>10) achieved in short integration times (<1 min). The observations from a validation deployment to the Shoals Marine Lab on Appledore Island, ME are presented and are broadly consistent with in situ observations from European Coastal Sites. Mixing ratios ranged from the instrumental detection limit (<1 pptv) to 10 pptv. These data represent the first in situ point measurements of IO in North America.

The development and deployment of a ground-based, laser-induced fluorescence instrument for the in situ detection of iodine monoxide radicals

International Nuclear Information System (INIS)

Thurlow, M. E.; Hannun, R. A.; Lapson, L. B.; Anderson, J. G.; Co, D. T.; O'Brien, A. S.; Hanisco, T. F.

2014-01-01

High abundances of iodine monoxide (IO) are known to exist and to participate in local photochemistry of the marine boundary layer. Of particular interest are the roles IO plays in the formation of new particles in coastal marine environments and in depletion episodes of ozone and mercury in the Arctic polar spring. This paper describes a ground-based instrument that measures IO at mixing ratios less than one part in 10 12 . The IO radical is measured by detecting laser-induced fluorescence at wavelengths longer that 500 nm. Tunable visible light is used to pump the A 2 Π 3/2 (v ′ = 2) ← X 2 Π 3/2 (v ″ = 0) transition of IO near 445 nm. The laser light is produced by a solid-state, Nd:YAG-pumped Ti:Sapphire laser at 5 kHz repetition rate. The laser-induced fluorescence instrument performs reliably with very high signal-to-noise ratios (>10) achieved in short integration times (<1 min). The observations from a validation deployment to the Shoals Marine Lab on Appledore Island, ME are presented and are broadly consistent with in situ observations from European Coastal Sites. Mixing ratios ranged from the instrumental detection limit (<1 pptv) to 10 pptv. These data represent the first in situ point measurements of IO in North America

Tunable laser optics

CERN Document Server

Duarte, FJ

2015-01-01

This Second Edition of a bestselling book describes the optics and optical principles needed to build lasers. It also highlights the optics instrumentation necessary to characterize laser emissions and focuses on laser-based optical instrumentation. The book emphasizes practical and utilitarian aspects of relevant optics including the essential theory. This revised, expanded, and improved edition contains new material on tunable lasers and discusses relevant topics in quantum optics.

Effect of laser pulse energies in laser induced breakdown spectroscopy in double-pulse configuration

International Nuclear Information System (INIS)

Benedetti, P.A.; Cristoforetti, G.; Legnaioli, S.; Palleschi, V.; Pardini, L.; Salvetti, A.; Tognoni, E.

2005-01-01

In this paper, the effect of laser pulse energy on double-pulse laser induced breakdown spectroscopy signal is studied. In particular, the energy of the first pulse has been changed, while the second pulse energy is held fixed. A systematic study of the laser induced breakdown spectroscopy signal dependence on the interpulse delay is performed, and the results are compared with the ones obtained with a single laser pulse of energy corresponding to the sum of the two pulses. At the same time, the crater formed at the target surface is studied by video-confocal microscopy, and the variation in crater dimensions is correlated to the enhancement of the laser induced breakdown spectroscopy signal. The results obtained are consistent with the interpretation of the double-pulse laser induced breakdown spectroscopy signal enhancement in terms of the changes in ambient gas pressure produced by the shock wave induced by the first laser pulse

Using Photoshop with images created by a confocal system.

Science.gov (United States)

Sedgewick, Jerry

2014-01-01

Many pure colors and grayscales tones that result from confocal imaging are not reproducible to output devices, such as printing presses, laptop projectors, and laser jet printers. Part of the difficulty in predicting the colors and tones that will reproduce lies in both the computer display, and in the display of unreproducible colors chosen for fluorophores. The use of a grayscale display for confocal channels and a LUT display to show saturated (clipped) tonal values aids visualization in the former instance and image integrity in the latter. Computer monitors used for post-processing in order to conform the image to the output device can be placed in darkened rooms, and the gamma for the display can be set to create darker shadow regions, and to control the display of color. These conditions aid in visualization of images so that blacks are set to grayer values that are more amenable to faithful reproduction. Preferences can be set in Photoshop for consistent display of colors, along with other settings to optimize use of memory. The Info window is opened so that tonal information can be shown via readouts. Images that are saved as indexed color are converted to grayscale or RGB Color, 16-bit is converted to 8-bit when desired, and colorized images from confocal software is returned to grayscale and re-colorized according to presented methods so that reproducible colors are made. Images may also be sharpened and noise may be reduced, or more than one image layered to show colocalization according to specific methods. Images are then converted to CMYK (Cyan, Magenta, Yellow and Black) for consequent assignment of pigment percentages for printing presses. Changes to single images and multiple images from image stacks are automated for efficient and consistent image processing changes. Some additional changes are done to those images destined for 3D visualization to better separate regions of interest from background. Files are returned to image stacks, saved and

Confocal Microscopy of Unfixed Breast Needle Core Biopsies: A Comparison to Fixed and Stained Sections

Directory of Open Access Journals (Sweden)

Zavislan James M

2009-08-01

Full Text Available Abstract Background Needle core biopsy, often in conjunction with ultrasonic or stereotactic guided techniques, is frequently used to diagnose breast carcinoma in women. Confocal scanning laser microscopy (CSLM is a technology that provides real-time digital images of tissues with cellular resolution. This paper reports the progress in developing techniques to rapidly screen needle core breast biopsy and surgical specimens at the point of care. CSLM requires minimal tissue processing and has the potential to reduce the time from excision to diagnosis. Following imaging, specimens can still be submitted for standard histopathological preparation. Methods Needle core breast specimens from 49 patients were imaged at the time of biopsy. These lesions had been characterized under the Breast Imaging Reporting And Data System (BI-RADS as category 3, 4 or 5. The core biopsies were imaged with the CSLM before fixation. Samples were treated with 5% citric acid and glycerin USP to enhance nuclear visibility in the reflectance confocal images. Immediately following imaging, the specimens were fixed in buffered formalin and submitted for histological processing and pathological diagnosis. CSLM images were then compared to the standard histology. Results The pathologic diagnoses by standard histology were 7 invasive ductal carcinomas, 2 invasive lobular carcinomas, 3 ductal carcinomas in-situ (CIS, 21 fibrocystic changes/proliferative conditions, 9 fibroadenomas, and 5 other/benign; two were excluded due to imaging difficulties. Morphologic and cellular features of benign and cancerous lesions were identified in the confocal images and were comparable to standard histologic sections of the same tissue. Conclusion CSLM is a technique with the potential to screen needle core biopsy specimens in real-time. The confocal images contained sufficient information to identify stromal reactions such as fibrosis and cellular proliferations such as intra-ductal and

Confocal Microscopy of Unfixed Breast Needle Core Biopsies: A Comparison to Fixed and Stained Sections

Science.gov (United States)

2009-01-01

Background Needle core biopsy, often in conjunction with ultrasonic or stereotactic guided techniques, is frequently used to diagnose breast carcinoma in women. Confocal scanning laser microscopy (CSLM) is a technology that provides real-time digital images of tissues with cellular resolution. This paper reports the progress in developing techniques to rapidly screen needle core breast biopsy and surgical specimens at the point of care. CSLM requires minimal tissue processing and has the potential to reduce the time from excision to diagnosis. Following imaging, specimens can still be submitted for standard histopathological preparation. Methods Needle core breast specimens from 49 patients were imaged at the time of biopsy. These lesions had been characterized under the Breast Imaging Reporting And Data System (BI-RADS) as category 3, 4 or 5. The core biopsies were imaged with the CSLM before fixation. Samples were treated with 5% citric acid and glycerin USP to enhance nuclear visibility in the reflectance confocal images. Immediately following imaging, the specimens were fixed in buffered formalin and submitted for histological processing and pathological diagnosis. CSLM images were then compared to the standard histology. Results The pathologic diagnoses by standard histology were 7 invasive ductal carcinomas, 2 invasive lobular carcinomas, 3 ductal carcinomas in-situ (CIS), 21 fibrocystic changes/proliferative conditions, 9 fibroadenomas, and 5 other/benign; two were excluded due to imaging difficulties. Morphologic and cellular features of benign and cancerous lesions were identified in the confocal images and were comparable to standard histologic sections of the same tissue. Conclusion CSLM is a technique with the potential to screen needle core biopsy specimens in real-time. The confocal images contained sufficient information to identify stromal reactions such as fibrosis and cellular proliferations such as intra-ductal and infiltrating carcinoma, and

Intercomparison of Terrestrial Laser Scanning Instruments for Assessing Forested Ecosystems: A Brisbane Field Experiment

Science.gov (United States)

Armston, J.; Newnham, G.; Strahler, A. H.; Schaaf, C.; Danson, M.; Gaulton, R.; Zhang, Z.; Disney, M.; Sparrow, B.; Phinn, S. R.; Schaefer, M.; Burt, A.; Counter, S.; Erb, A.; Goodwin, N.; Hancock, S.; Howe, G.; Johansen, K.; Li, Z.; Lollback, G.; Martel, J.; Muir, J.; Paynter, I.; Saenz, E.; Scarth, P.; Tindall, D.; Walker, L.; Witte, C.; Woodgate, W.; Wu, S.

2013-12-01

During 28th July - 3rd August, 2013, an international group of researchers brought five terrestrial laser scanners (TLS) to long-term monitoring plots in three eucalyptus-dominated woodland sites near Brisbane, Queensland, Australia, to acquire scans at common locations for calibration and intercomparison.They included: DWEL - a dual-wavelength full-waveform laser scanner (Boston U., U. Massachusetts Lowell, U. Massachusetts Boston, USA) SALCA - a dual-wavelength full-waveform laser scanner (U. Salford, UK) CBL - a canopy biomass lidar, a small ultraportable low-cost multiple discrete return scanner (U. Massachusetts Boston, USA) Riegl VZ400 - a survey-grade commercial waveform scanner (Queensland Government and TERN, U. Queensland, Australia) FARO Focus 3D - a lightweight commercial phase-shift ranging laser scanner (U. Southern Queensland) Two plots were scanned at Karawatha Forest Park, a Terrestrial Ecosystem Research Network (TERN) Supersite, and one plot at D'Aguilar National Park. At each 50 x 100 m plot, a center scan point was surrounded by four scan points located 25 m away in a cross pattern allowing for 3-D reconstructions of scan sites in the form of point clouds. At several center points, multiple instrument configurations (i.e. different beam divergence, angular resolution, pulse rate) were acquired to test the impact of instrument specifications on separation of woody and non-woody materials and estimation of vegetation structure parameters. Three-dimensional Photopoint photographic panoramas were also acquired, providing reconstructions of stems in the form of point clouds using photogrammetric correlation methods. Calibrated reflectance targets were also scanned to compare instrument geometric and radiometric performance. Ancillary data included hemispherical photos, TRAC LAI/clumping measurements, spectra of leaves, bark, litter, and other target components. Wet and dry leaf weights determined water content. Planned intercomparison topics and

Cell Attachment Following Instrumentation with Titanium and Plastic Instruments, Diode Laser, and Titanium Brush on Titanium, Titanium-Zirconium, and Zirconia Surfaces.

Science.gov (United States)

Lang, Melissa S; Cerutis, D Roselyn; Miyamoto, Takanari; Nunn, Martha E

2016-01-01

The aim of this study was to evaluate the surface characteristics and gingival fibroblast adhesion of disks composed of implant and abutment materials following brief and repeated instrumentation with instruments commonly used in procedures for implant maintenance, stage-two implant surgery, and periimplantitis treatment. One hundred twenty disks (40 titanium, 40 titaniumzirconium, 40 zirconia) were grouped into treatment categories of instrumentation by plastic curette, titanium curette, diode microlaser, rotary titanium brush, and no treatment. Twenty strokes were applied to half of the disks in the plastic and titanium curette treatment categories, while half of the disks received 100 strokes each to simulate implant maintenance occurring on a repetitive basis. Following analysis of the disks by optical laser profilometry, disks were cultured with human gingival fibroblasts. Cell counts were conducted from scanning electron microscopy (SEM) images. Differences in surface roughness across all instruments tested for zirconia disks were negligible, while both titanium disks and titaniumzirconium disks showed large differences in surface roughness across the spectrum of instruments tested. The rotary titanium brush and the titanium curette yielded the greatest overall mean surface roughness, while the plastic curette yielded the lowest mean surface roughness. The greatest mean cell counts for each disk type were as follows: titanium disks with plastic curettes, titanium-zirconium disks with titanium curettes, and zirconia disks with the diode microlaser. Repeated instrumentation did not result in cumulative changes in surface roughness of implant materials made of titanium, titanium-zirconium, or zirconia. Instrumentation with plastic implant curettes on titanium and zirconia surfaces appeared to be more favorable than titanium implant curettes in terms of gingival fibroblast attachment on these surfaces.

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.

An overview of the laser ranging method of space laser altimeter

Science.gov (United States)

Zhou, Hui; Chen, Yuwei; Hyyppä, Juha; Li, Song

2017-11-01

Space laser altimeter is an active remote sensing instrument to measure topographic map of Earth, Moon and planetary. The space laser altimeter determines the range between the instrument and laser footprint by measuring round trip time of laser pulse. The return pulse reflected from ground surface is gathered by the receiver of space laser altimeter, the pulsewidth and amplitude of which are changeable with the variability of the ground relief. Meantime, several kinds of noise overlapped on the return pulse signal affect its signal-to-noise ratio. To eliminate the influence of these factors that cause range walk and range uncertainty, the reliable laser ranging methods need to be implemented to obtain high-precision range results. Based on typical space laser altimeters in the past few decades, various ranging methods are expounded in detail according to the operational principle of instruments and timing method. By illustrating the concrete procedure of determining time of flight of laser pulse, this overview provides the comparison of the employed technologies in previous and undergoing research programs and prospect innovative technology for space laser altimeters in future.

Cell volume and geometric parameters determination in living cells using confocal microscopy and 3D reconstruction

OpenAIRE

sprotocols

2015-01-01

Authors: David Hevia, Aida Rodriguez-Garcia, Marta Alonso-Gervós, Isabel Quirós-González, Henar M Cimadevilla, Carmen Gómez-Cordovés, Rosa M Sainz & Juan C Mayo ### Abstract The protocol reported here describes a simple, easy, fast and reproducible method aimed to know the geometric parameters of living cells based on confocal laser scanning microscopy combined with 3D reconstruction software. Briefly, the method is based on intrinsic fluorescence properties of acridine orange (AO), a...

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

The OSIRIS-REx Laser Altimeter (OLA) Investigation and Instrument

Science.gov (United States)

Daly, M. G.; Barnouin, O. S.; Dickinson, C.; Seabrook, J.; Johnson, C. L.; Cunningham, G.; Haltigin, T.; Gaudreau, D.; Brunet, C.; Aslam, I.; Taylor, A.; Bierhaus, E. B.; Boynton, W.; Nolan, M.; Lauretta, D. S.

2017-10-01

The Canadian Space Agency (CSA) has contributed to the Origins Spectral Interpretation Resource Identification Security-Regolith Explorer (OSIRIS-REx) spacecraft the OSIRIS-REx Laser Altimeter (OLA). The OSIRIS-REx mission will sample asteroid 101955 Bennu, the first B-type asteroid to be visited by a spacecraft. Bennu is thought to be primitive, carbonaceous, and spectrally most closely related to CI and/or CM meteorites. As a scanning laser altimeter, the OLA instrument will measure the range between the OSIRIS-REx spacecraft and the surface of Bennu to produce digital terrain maps of unprecedented spatial scales for a planetary mission. The digital terrain maps produced will measure ˜7 cm per pixel globally, and ˜3 cm per pixel at specific sample sites. In addition, OLA data will be used to constrain and refine the spacecraft trajectories. Global maps and highly accurate spacecraft trajectory estimates are critical to infer the internal structure of the asteroid. The global and regional maps also are key to gain new insights into the surface processes acting across Bennu, which inform the selection of the OSIRIS-REx sample site. These, in turn, are essential for understanding the provenance of the regolith sample collected by the OSIRIS-REx spacecraft. The OLA data also are important for quantifying any hazards near the selected OSIRIS-REx sample site and for evaluating the range of tilts at the sampling site for comparison against the capabilities of the sample acquisition device.

Systematic review of the adjunctive use of diode and Nd:YAG lasers for nonsurgical periodontal instrumentation.

Science.gov (United States)

Roncati, Marisa; Gariffo, Annalisa

2014-04-01

The aims of this study were (1) to conduct a literature search and systematically evaluate the additional therapeutic effects of pulsed Nd:YAG or diode laser use in patients with periodontitis, (2) to assess evidence supporting the additional benefit of laser-mediated periodontal treatment in conjunction with scaling and root planning (SRP) (not as monotherapy), and (3) to interpret the evidence presented in retrieved publications. Opinions about the additional use of diode lasers in the nonsurgical treatment of plaque-induced periodontal lesions are conflicting. The April 2011 American Academy of Periodontology's "Statement on the Efficacy of Lasers in the Non-Surgical Treatment of Inflammatory Periodontal Disease" asserted that the use of a laser as monotherapy or in addition to nonsurgical periodontal instrumentation conveyed no advantage. After initial screening, 23/77 potentially relevant articles and abstracts identified through electronic and manual searches of the MEDLINE(®)/PubMed database and the Cochrane Central Register of Controlled Trials (1990-2012) were included in this review. A meta-analysis could be performed. The results indicate that Nd:YAG or diode laser, used in an adjunctive capacity to SRP, may provide some additional benefit, in 6 month studies, compared with mechanical debridement. The results show the adjunctive benefits that diode laser treatment can provide when it is used as an adjunct to nonsurgical periodontal treatment in adults with chronic periodontitis. Further long-term, well-designed, parallel randomized clinical trials are needed to assess the effectiveness of the adjunctive use of the diode laser, as well as the appropriate dosimetry and laser settings.

Prototype of a Laser-Induced Fluorescence Ground-Based Instrument for Measurements of Atmospheric Iodine Monoxide (IO)

Science.gov (United States)

Thurlow, M. E.; Co, D. T.; Hanisco, T. F.; Lapson, L. B.; Anderson, J. G.

2008-12-01

High abundances of iodine monoxide (IO) are known to exist and to participate in local photochemistry of the marine boundary layer: (1) IO participates in depletion episodes of O3 and in the removal of mercury in the Arctic polar spring by enhancing atomic Br mixing ratios. Recent observations and computer simulations suggest that mercury sequestration is closely tied to halogen photochemistry and that gaseous atomic Hg depletion can be enhanced significantly by the presence of small amounts of iodine-containing compounds. (2) IO and higher- order iodine oxides are involved in the formation of new particles in coastal marine environments. Studies using smog chamber experiments simulating coastal atmospheric conditions have demonstrated that new particles can form from condensable iodine-containing vapors and that their concentrations over the open ocean are sufficient to influence marine particle formation. (3) IO has also been shown to affect the oxidizing capacity of the troposphere by altering the partitioning of NO2/NO and HO2/HO and by activating chlorine and bromine in sea salt aerosols. In the stratosphere, these same processes can lead to enhanced ozone loss rates. Detailed photochemical models that include iodine photochemistry, however, are hampered by the lack of observational data. The distribution of IO in vertical, horizontal, and temporal coordinates is unknown, so the impact of IO on global photochemistry cannot be predicted. The resolution of these important scientific issues requires an in situ IO instrument. A fully functional nanosecond Nd:YAG-pumped Ti:Sapphire laser system and a prototype IO ground-based instrument have been built in our lab. With the current setup, the laser system was situated 10 m from the field station, and the laser light was coupled via an optical fiber. With the use of highly efficient fluorescence detection optics and photon counting techniques, sensitivities of better than 0.1 ppt in 1 s for IO was achieved in the

Instrumentation for low noise nanopore-based ionic current recording under laser illumination

Science.gov (United States)

Roelen, Zachary; Bustamante, José A.; Carlsen, Autumn; Baker-Murray, Aidan; Tabard-Cossa, Vincent

2018-01-01

We describe a nanopore-based optofluidic instrument capable of performing low-noise ionic current recordings of individual biomolecules under laser illumination. In such systems, simultaneous optical measurements generally introduce significant parasitic noise in the electrical signal, which can severely reduce the instrument sensitivity, critically hindering the monitoring of single-molecule events in the ionic current traces. Here, we present design rules and describe simple adjustments to the experimental setup to mitigate the different noise sources encountered when integrating optical components to an electrical nanopore system. In particular, we address the contributions to the electrical noise spectra from illuminating the nanopore during ionic current recording and mitigate those effects through control of the illumination source and the use of a PDMS layer on the SiNx membrane. We demonstrate the effectiveness of our noise minimization strategies by showing the detection of DNA translocation events during membrane illumination with a signal-to-noise ratio of ˜10 at 10 kHz bandwidth. The instrumental guidelines for noise minimization that we report are applicable to a wide range of nanopore-based optofluidic systems and offer the possibility of enhancing the quality of synchronous optical and electrical signals obtained during single-molecule nanopore-based analysis.

Time- and Space-Resolved Spectroscopic Investigation on Pi-Conjugated Nanostructures - 2

Science.gov (United States)

2016-01-12

Defocused wide-field fluorescence (DWFI) microscopy suggests that molecular heterogeneities and flexibilities clearly depend on ring size and that site... Confocal   Microscopy   Setup Wild‐field  Microscopy   Setup Femtosecond Z‐scan  experiment Setup Figure 3. Instruments of Time- and space-resolved...approved for public release. 3. Space-Resolved Laser Spectroscopy - Confocal Microscopy - Wild-field Microscopy 4. Non-Linear Spectroscopy

Fault localization and analysis in semiconductor devices with optical-feedback infrared confocal microscopy

International Nuclear Information System (INIS)

Sarmiento, Raymund; Cemine, Vernon Julius; Tagaca, Imee Rose; Salvador, Arnel; Mar Blanca, Carlo; Saloma, Caesar

2007-01-01

We report on a cost-effective optical setup for characterizing light-emitting semiconductor devices with optical-feedback confocal infrared microscopy and optical beam-induced resistance change.We utilize the focused beam from an infrared laser diode to induce local thermal resistance changes across the surface of a biased integrated circuit (IC) sample. Variations in the multiple current paths are mapped by scanning the IC across the focused beam. The high-contrast current maps allow accurate differentiation of the functional and defective sites, or the isolation of the surface-emittingp-i-n devices in the IC. Optical beam-induced current (OBIC) is not generated since the incident beam energy is lower than the bandgap energy of the p-i-n device. Inhomogeneous current distributions in the IC become apparent without the strong OBIC background. They are located at a diffraction-limited resolution by referencing the current maps against the confocal reflectance image that is simultaneously acquired via optical-feedback detection. Our technique permits the accurate identification of metal and semiconductor sites as well as the classification of different metallic structures according to thickness, composition, or spatial inhomogeneity

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.

Confocal Endomicroscopy of Colorectal Polyps

Directory of Open Access Journals (Sweden)

Vivian M. Ussui

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 micron scale via endoscopes. CLE has the potential to be a disruptive technology in that it can change the current algorithms that depend on biopsy to perform surveillance of high-risk conditions. Furthermore, it allows on-table decision making that has the potential to guide therapy in real time and reduce the need for repeated procedures. CLE and related technologies are often termed “virtual biopsy” as they simulate the images seen in traditional histology. However, the imaging of living tissue allows more than just pragmatic convenience; it also allows imaging of living tissue such as active capillary circulation, cellular death, and vascular and endothelial translocation, thus extending beyond what is capable in traditional biopsy. Immediate potential applications of CLE are to guide biopsy sampling in Barrett's esophagus and inflammatory bowel disease surveillance, evaluation of colorectal polyps, and intraductal imaging of the pancreas and bile duct. Data on these applications is rapidly emerging, and more is needed to clearly demonstrate the optimal applications of CLE. In this paper, we will focus on the role of CLE as applied to colorectal polyps detected during colonoscopy.

Dual filtered backprojection for micro-rotation confocal microscopy

International Nuclear Information System (INIS)

Laksameethanasan, Danai; Brandt, Sami S; Renaud, Olivier; Shorte, Spencer L

2009-01-01

Micro-rotation confocal microscopy is a novel optical imaging technique which employs dielectric fields to trap and rotate individual cells to facilitate 3D fluorescence imaging using a confocal microscope. In contrast to computed tomography (CT) where an image can be modelled as parallel projection of an object, the ideal confocal image is recorded as a central slice of the object corresponding to the focal plane. In CT, the projection images and the 3D object are related by the Fourier slice theorem which states that the Fourier transform of a CT image is equal to the central slice of the Fourier transform of the 3D object. In the micro-rotation application, we have a dual form of this setting, i.e. the Fourier transform of the confocal image equals the parallel projection of the Fourier transform of the 3D object. Based on the observed duality, we present here the dual of the classical filtered back projection (FBP) algorithm and apply it in micro-rotation confocal imaging. Our experiments on real data demonstrate that the proposed method is a fast and reliable algorithm for the micro-rotation application, as FBP is for CT application

Femtosecond laser inscribed cladding waveguides in Nd:YAG ceramics: fabrication, fluorescence imaging and laser performance.

Science.gov (United States)

Liu, Hongliang; Jia, Yuechen; Vázquez de Aldana, Javier Rodríguez; Jaque, Daniel; Chen, Feng

2012-08-13

We report on the fabrication of depressed cladding waveguide lasers in Nd:YAG (neodymium doped yttrium aluminum garnet, Nd:Y3Al5O12) ceramics microstructured by femtosecond laser pulses. Full control over the confined light spatial distribution is demonstrated by the fabrication of high contrast waveguides with hexagonal, circular and trapezoidal configurations. The confocal fluorescence measurements of the waveguides reveal that the original luminescence features of Nd3+ ions are well-preserved in the waveguide regions. Under optical pump at 808 nm, cladding waveguides showed continuous wave efficient laser oscillation. The maximum output power obtained at 1064.5 nm is ~181 mW with a slope efficiency as high as 44%, which suggests that the fabricated Nd:YAG ceramic waveguides are promising candidates for efficient integrated laser sources.

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.

Microfabrication of biomaterials by the sub-ps laser-induced forward transfer process

International Nuclear Information System (INIS)

Karaiskou, A.; Zergioti, I.; Fotakis, C.; Kapsetaki, M.; Kafetzopoulos, D.

2003-01-01

The precise fabrication of micro-dimensioned patterns of biomaterials by the laser microprinting technique using a sub-ps UV laser is described. An ultrashort UV laser has been used to transfer the biomaterial, with low angular divergence, and deposit it onto the substrate with minimum spread and high spatial resolution. The laser-transferred features of 100 μmx100 μm size have been studied by means of scanning electron microscopy and scanning laser confocal fluorescence microscopy. The analysis of DNA and protein microarrays provides an excellent tool to expand our knowledge of genome functions

Synchrotron light sources and free-electron lasers accelerator physics, instrumentation and science applications

CERN Document Server

Khan, Shaukat; Schneider, Jochen; Hastings, Jerome

2016-01-01

Hardly any other discovery of the nineteenth century did have such an impact on science and technology as Wilhelm Conrad Röntgen’s seminal find of the X-rays. X-ray tubes soon made their way as excellent instruments for numerous applications in medicine, biology, materials science and testing, chemistry and public security. Developing new radiation sources with higher brilliance and much extended spectral range resulted in stunning developments like the electron synchrotron and electron storage ring and the freeelectron laser. This handbook highlights these developments in fifty chapters. The reader is given not only an inside view of exciting science areas but also of design concepts for the most advanced light sources. The theory of synchrotron radiation and of the freeelectron laser, design examples and the technology basis are presented. The handbook presents advanced concepts like seeding and harmonic generation, the booming field of Terahertz radiation sources and upcoming brilliant light sources dri...

WE-FG-BRA-04: A Portable Confocal Microscope to Image Live Cell Damage Response Induced by Therapeutic Radiation

Energy Technology Data Exchange (ETDEWEB)

McFadden, C; Flint, D; Grosshans, D; Sawakuchi, G [The University of Texas MD Anderson Cancer Center, Houston, TX (United States); Sadetaporn, D [The University of Texas MD Anderson Cancer Center, Houston, TX (United States); Rice University, Houston, TX (United States); Asaithamby, A [UT Southwestern Medical Center, Dallas, TX (United States)

2016-06-15

Purpose: To construct a custom and portable fluorescence confocal laser-scanning microscope (FCLSM) that can be placed in the path of therapeutic radiation beams to study real-time radiation-induced damage response in live cells. Methods: We designed and constructed a portable FCLSM with three laser diodes for excitation (405, 488, and 635 nm). An objective lens focuses the excitation light and collects fluorescence from the sample. A pair of galvanometer mirrors scans/collects the laser beam/fluorescence along the focal plane (x/y-directions). A stepper motor stage scans in the axial direction and positions the x/y of the image field. Barrier filters and dichroic mirrors are used to route the spectral emission bands to the appropriate photodetector. An avalanche photodiode collects near-infrared fluorescence; a photodiode collects back-reflected 635 nm light; and a photomultiplier tube collects green fluorescence in the range of eGFP/eYFP. A 200-µm diameter pinhole was used to implement the confocal geometry for near-infrared and red channels and a 150-µm diameter pinhole for the green channel. Data acquisition and system control were achieved using a high-throughput data acquisition card. In-house software developed in LabVIEW was used to control the hardware, collect data from the photodetectors and reconstruct the confocal images. Results: 6 frames/s can be acquired for a 25 µm{sup 2} (128×128 pixels) field of view, visualizing the entire volume of the cell nucleus (∼10 µm depth) in <10 s. To demonstrate the usefulness of our FCLSM, we imaged gold nanoshells in live cells, radiation-induced damage in fibrosarcoma cells expressing eGFP tagged to a DNA repair protein, and neurons expressing eGFP. The system can also image particle tracks in fluorescent nuclear track detectors. Conclusion: We developed a versatile and portable FCLSM that allows radiobiology studies in live cells exposed to therapeutic radiation. The FCLSM can be placed in any vertical beam

WE-FG-BRA-04: A Portable Confocal Microscope to Image Live Cell Damage Response Induced by Therapeutic Radiation

International Nuclear Information System (INIS)

McFadden, C; Flint, D; Grosshans, D; Sawakuchi, G; Sadetaporn, D; Asaithamby, A

2016-01-01

Purpose: To construct a custom and portable fluorescence confocal laser-scanning microscope (FCLSM) that can be placed in the path of therapeutic radiation beams to study real-time radiation-induced damage response in live cells. Methods: We designed and constructed a portable FCLSM with three laser diodes for excitation (405, 488, and 635 nm). An objective lens focuses the excitation light and collects fluorescence from the sample. A pair of galvanometer mirrors scans/collects the laser beam/fluorescence along the focal plane (x/y-directions). A stepper motor stage scans in the axial direction and positions the x/y of the image field. Barrier filters and dichroic mirrors are used to route the spectral emission bands to the appropriate photodetector. An avalanche photodiode collects near-infrared fluorescence; a photodiode collects back-reflected 635 nm light; and a photomultiplier tube collects green fluorescence in the range of eGFP/eYFP. A 200-µm diameter pinhole was used to implement the confocal geometry for near-infrared and red channels and a 150-µm diameter pinhole for the green channel. Data acquisition and system control were achieved using a high-throughput data acquisition card. In-house software developed in LabVIEW was used to control the hardware, collect data from the photodetectors and reconstruct the confocal images. Results: 6 frames/s can be acquired for a 25 µm 2 (128×128 pixels) field of view, visualizing the entire volume of the cell nucleus (∼10 µm depth) in <10 s. To demonstrate the usefulness of our FCLSM, we imaged gold nanoshells in live cells, radiation-induced damage in fibrosarcoma cells expressing eGFP tagged to a DNA repair protein, and neurons expressing eGFP. The system can also image particle tracks in fluorescent nuclear track detectors. Conclusion: We developed a versatile and portable FCLSM that allows radiobiology studies in live cells exposed to therapeutic radiation. The FCLSM can be placed in any vertical beam line

In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy

Institute of Scientific and Technical Information of China (English)

Martin Goetz; Beena Memadathil; Stefan Biesterfeld; Constantin Schneider; Sebastian Gregor; Peter R Galle; Markus F Neurath; Ralf Kiesslich

2007-01-01

AIM: To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents.METHODS: A novel rigid confocal probe (diameter 7 mm) was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation.Light emission was detected at 505 to 750 nm. The field of view was 475 μm × 475 μm. Optical slice thickness was 7 μm with a lateral resolution of 0.7 μm. Subsurface serial images at different depths (surface to 250 μm)were generated in real time at 1024 × 1024 pixels (0.8 frames/s) by placing the probe onto the tissue in gentle,stable contact. Tissue specimens were sampled for histopathological correlation.RESULTS: The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice.Real time microscopic imaging with the confocal minimicroscopy probe was easy to achieve. The different staining protocols (fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin) each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging.CONCLUSION: Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures.The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients.

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.

An electronically tunable ultrafast laser source applied to fluorescence imaging and fluorescence lifetime imaging microscopy

International Nuclear Information System (INIS)

Dunsby, C; Lanigan, P M P; McGinty, J; Elson, D S; Requejo-Isidro, J; Munro, I; Galletly, N; McCann, F; Treanor, B; Oenfelt, B; Davis, D M; Neil, M A A; French, P M W

2004-01-01

Fluorescence imaging is used widely in microscopy and macroscopic imaging applications for fields ranging from biomedicine to materials science. A critical component for any fluorescence imaging system is the excitation source. Traditionally, wide-field systems use filtered thermal or arc-generated white light sources, while point scanning confocal microscope systems require spatially coherent (point-like) laser sources. Unfortunately, the limited range of visible wavelengths available from conventional laser sources constrains the design and usefulness of fluorescent probes in confocal microscopy. A 'hands-off' laser-like source, electronically tunable across the visible spectrum, would be invaluable for fluorescence imaging and provide new opportunities, e.g. automated excitation fingerprinting and in situ measurement of excitation cross-sections. Yet more information can be obtained using fluorescence lifetime imaging (FLIM), which requires that the light source be pulsed or rapidly modulated. We show how a white light continuum, generated by injecting femtosecond optical radiation into a micro-structured optical fibre, coupled with a simple prism-based tunable filter arrangement, can fulfil all these roles as a continuously electronically tunable (435-1150 nm) visible ultrafast light source in confocal, wide-field and FLIM systems

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.

A comparative scanning electron microscopy study between hand instrument, ultrasonic scaling and erbium doped:Yttirum aluminum garnet laser on root surface: A morphological and thermal analysis

Directory of Open Access Journals (Sweden)

Mitul Kumar Mishra

2013-01-01

Full Text Available Background and Objectives: Scaling and root planing is one of the most commonly used procedures for the treatment of periodontal diseases. Removal of calculus using conventional hand instruments is incomplete and rather time consuming. In search of more efficient and less difficult instrumentation, investigators have proposed lasers as an alternative or as adjuncts to scaling and root planing. Hence, the purpose of the present study was to evaluate the effectiveness of erbium doped: Yttirum aluminum garnet (Er:YAG laser scaling and root planing alone or as an adjunct to hand and ultrasonic instrumentation. Subjects and Methods: A total of 75 freshly extracted periodontally involved single rooted teeth were collected. Teeth were randomly divided into five treatment groups having 15 teeth each: Hand scaling only, ultrasonic scaling only, Er:YAG laser scaling only, hand scaling + Er:YAG laser scaling and ultrasonic scaling + Er:YAG laser scaling. Specimens were subjected to scanning electron microscopy and photographs were evaluated by three examiners who were blinded to the study. Parameters included were remaining calculus index, loss of tooth substance index, roughness loss of tooth substance index, presence or absence of smear layer, thermal damage and any other morphological damage. Results: Er:YAG laser treated specimens showed similar effectiveness in calculus removal to the other test groups whereas tooth substance loss and tooth surface roughness was more on comparison with other groups. Ultrasonic treated specimens showed better results as compared to other groups with different parameters. However, smear layer presence was seen more with hand and ultrasonic groups. Very few laser treated specimens showed thermal damage and morphological change. Interpretation and Conclusion: In our study, ultrasonic scaling specimen have shown root surface clean and practically unaltered. On the other hand, hand instrument have produced a plane surface

Fluorescence confocal endomicroscopy in biological imaging

Science.gov (United States)

Delaney, Peter; Thomas, Steven; Allen, John; McLaren, Wendy; Murr, Elise; Harris, Martin

2007-02-01

In vivo fluorescence microscopic imaging of biological systems in human disease states and animal models is possible with high optical resolution and mega pixel point-scanning performance using optimised off-the-shelf turn-key devices. There are however various trade-offs between tissue access and instrument performance when miniaturising in vivo microscopy systems. A miniature confocal scanning technology that was developed for clinical human endoscopy has been configured into a portable device for direct hand-held interrogation of living tissue in whole animal models (Optiscan FIVE-1 system). Scanning probes of 6.3mm diameter with a distal tip diameter of 5.0mm were constructed either in a 150mm length for accessible tissue, or a 300mm probe for laparoscopic interrogation of internal tissues in larger animal models. Both devices collect fluorescence confocal images (excitation 488 nm; emission >505 or >550 nm) comprised of 1024 x 1204 sampling points/image frame, with lateral resolution 0.7um; axial resolution 7um; FOV 475 x 475um. The operator can dynamically control imaging depth from the tissue surface to approx 250um in 4um steps via an internally integrated zaxis actuator. Further miniaturisation is achieved using an imaging contact probe based on scanning the proximal end of a high-density optical fibre bundle (~30,000 fibres) of small animal organs, albeit at lower resolution (30,000 sampling points/image). In rodent models, imaging was performed using various fluorescent staining protocols including fluorescently labelled receptor ligands, labelled antibodies, FITC-dextrans, vital dyes and labelled cells administered topically or intravenously. Abdominal organs of large animals were accessed laparoscopically and contrasted using i.v. fluorescein-sodium. Articular cartilage of sheep and pigs was fluorescently stained with calcein-AM or fluorescein. Surface and sub-surface cellular and sub-cellular details could be readily visualised in vivo at high

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

Using STED and ELSM confocal microscopy for a better knowledge of fused silica polished glass interface

International Nuclear Information System (INIS)

Catrin, Rodolphe; Neauport, Jerome; Taroux, Daniel; Corbineau, Thomas; Cormont, Philippe; Maunier, Cedric; Legros, Philippe

2013-01-01

Characteristics and nature of close surface defects existing in fused silica polished optical surfaces were explored. Samples were deliberately scratched using a modified polishing process in presence of different fluorescent dyes. Various techniques including Epi-fluorescence Laser Scanning Mode (ELSM) or Stimulated Emission Depletion (STED) confocal microscopy were used to measure and quantify scratches that are sometimes embedded under the polished layer. We show using a nondestructive technique that depth of the modified region extends far below the surface. Moreover cracks of 120 nm width can be present ten micrometers below the surface. (authors)

Smartphone confocal microscopy for imaging cellular structures in human skin in vivo.

Science.gov (United States)

Freeman, Esther E; Semeere, Aggrey; Osman, Hany; Peterson, Gary; Rajadhyaksha, Milind; González, Salvador; Martin, Jeffery N; Anderson, R Rox; Tearney, Guillermo J; Kang, Dongkyun

2018-04-01

We report development of a low-cost smartphone confocal microscope and its first demonstration of in vivo human skin imaging. The smartphone confocal microscope uses a slit aperture and diffraction grating to conduct two-dimensional confocal imaging without using any beam scanning devices. Lateral and axial resolutions of the smartphone confocal microscope were measured as 2 and 5 µm, respectively. In vivo confocal images of human skin revealed characteristic cellular structures, including spinous and basal keratinocytes and papillary dermis. Results suggest that the smartphone confocal microscope has a potential to examine cellular details in vivo and may help disease diagnosis in resource-poor settings, where conducting standard histopathologic analysis is challenging.

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.

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.

Ex Vivo Confocal Spectroscopy of Autofluorescence in Age-Related Macular Degeneration.

Directory of Open Access Journals (Sweden)

Joel Kaluzny

Full Text Available We investigated the autofluorescence (AF signature of the microscopic features of retina with age-related macular degeneration (AMD using 488 nm excitation.The globes of four donors with AMD and four age-matched controls were embedded in paraffin and sectioned through the macula. Sections were excited using a 488 nm argon laser, and the AF emission was captured using a laser scanning confocal microscope (496-610 nm, 6 nm resolution. The data cubes were then analyzed to compare peak emission spectra between the AMD and the controls. Microscopic features, including individual lipofuscin and melanolipofuscin granules, Bruch's Membrane, as well macroscopic features, were considered.Overall, the AMD eyes showed a trend of blue-shifted emission peaks compared with the controls. These differences were statistically significant when considering the emission of the combined RPE/Bruch's Membrane across all the tissue cross-sections (p = 0.02.The AF signatures of ex vivo AMD RPE/BrM show blue-shifted emission spectra (488 nm excitation compared with the control tissue. The magnitude of these differences is small (~4 nm and highlights the potential challenges of detecting these subtle spectral differences in vivo.

Hypericin and pulsed laser therapy of squamous cell cancer in vitro.

Science.gov (United States)

Bublik, Michael; Head, Christian; Benharash, Peyman; Paiva, Marcos; Eshraghi, Adrian; Kim, Taiho; Saxton, Romaine

2006-06-01

This in vitro study compares continuous wave and pulsed laser light at longer wavelengths for activation of the phototoxic drug hypericin in human cancer cells. Two-photon pulsed laser light now allows high-resolution fluorescent imaging of cancer cells and should provide deeper tissue penetration with near infrared light for improved detection as well as phototoxicity in human tumors. Cultured Seoul National University (SNU)-1 tumor cells from a squamous cell carcinoma (SCC) were incubated with hypericin before photoirradiation at four laser wavelengths. Phototoxicity of hypericin sensitized SCC cells was measured by dimethyl thiazoldiphenyl (MTT) tetrazolium bromide cell viability assays and by confocal fluorescence microscopy via 532-nm and infrared two-photon pulsed laser light. Phototoxic response increased linearly with hypericin dose of 0.1-2 microM, light exposure time of 5-120 sec, and pulsed dye laser wavelengths of 514-593 nm. Light energy delivery for 50% cell phototoxicity (LD50) response was 9 joules at 514 nm, 3 joules at 550 nm, and less than 1 joule at the 593 nm hypericin light absorption maxima. Fluorescence confocal microscopy revealed membrane and perinuclear localization of hypericin in the SNU cells with membrane damage seen after excitation with visible 532 nm continuous wave light or two-photon 700-950 nm picosecond pulsed laser irradiation. Hypericin may be a powerful tumor targetting drug when combined with pulsed laser light in patients with recurrent head and neck SCC.

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)

Aorta Fluorescence Imaging by Using Confocal Microscopy

OpenAIRE

Wang, Chun-Yang; Tsai, Jui-che; Chuang, Ching-Cheng; Hsieh, Yao-Sheng; Sun, Chia-Wei

2011-01-01

The activated leukocyte attacked the vascular endothelium and the associated increase in VEcadherin number was observed in experiments. The confocal microscopic system with a prism-based wavelength filter was used for multiwavelength fluorescence measurement. Multiwavelength fluorescence imaging based on the VEcadherin within the aorta segment of a rat was achieved. The confocal microscopic system capable of fluorescence detection of cardiovascular tissue is a useful tool for measuring the bi...

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.

Fused oblique incidence reflectometry and confocal fluorescence microscopy

Science.gov (United States)

Risi, Matthew D.; Rouse, Andrew R.; Gmitro, Arthur F.

2011-03-01

Confocal microendoscopy provides real-time high resolution cellular level images via a minimally invasive procedure, but relies on exogenous fluorophores, has a relatively limited penetration depth (100 μm) and field of view (700 μm), and produces a high rate of detailed information to the user. A new catheter based multi-modal system has been designed that combines confocal imaging and oblique incidence reflectometry (OIR), which is a non-invasive method capable of rapidly extracting tissue absorption, μa, and reduced scattering, μ's, spectra from tissue. The system builds on previous developments of a custom slit-scan multi-spectral confocal microendoscope and is designed to rapidly switch between diffuse spectroscopy and confocal fluorescence imaging modes of operation. An experimental proof-of-principle catheter has been developed that consists of a fiber bundle for traditional confocal fluorescence imaging and a single OIR source fiber which is manually redirected at +/- 26 degrees. Diffusely scattered light from each orientation of the source fiber is collected via the fiber bundle, with a frame of data representing spectra collected at a range of distances from the OIR source point. Initial results with intralipid phantoms show good agreement to published data over the 550-650 nm spectral range. We successfully imaged and measured the optical properties of rodent cardiac muscle.

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

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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

Comparing phototoxicity during the development of a zebrafish craniofacial bone using confocal and light sheet fluorescence microscopy techniques.

Science.gov (United States)

Jemielita, Matthew; Taormina, Michael J; Delaurier, April; Kimmel, Charles B; Parthasarathy, Raghuveer

2013-12-01

The combination of genetically encoded fluorescent proteins and three-dimensional imaging enables cell-type-specific studies of embryogenesis. Light sheet microscopy, in which fluorescence excitation is provided by a plane of laser light, is an appealing approach to live imaging due to its high speed and efficient use of photons. While the advantages of rapid imaging are apparent from recent work, the importance of low light levels to studies of development is not well established. We examine the zebrafish opercle, a craniofacial bone that exhibits pronounced shape changes at early developmental stages, using both spinning disk confocal and light sheet microscopies of fluorescent osteoblast cells. We find normal and aberrant opercle morphologies for specimens imaged with short time intervals using light sheet and spinning disk confocal microscopies, respectively, under equivalent exposure conditions over developmentally-relevant time scales. Quantification of shapes reveals that the differently imaged specimens travel along distinct trajectories in morphological space. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Proceedings of the twenty fifth national laser symposium

International Nuclear Information System (INIS)

2016-01-01

The topics covered in this symposium are: laser materials, devices and components, nonlinear, quantum optics and atomic optics, ultrafast lasers and applications, physics and technology of lasers, lasers in nuclear science and technology, lasers in material science, laser plasma interaction, lasers in industry and defence, lasers in spectroscopy and applications, lasers in chemistry, biology and medicine, laser based instrumentation and electronics and instrumentation for lasers. Papers relevant to INIS are indexed separately

The laser, measuring instrument for plasmas

International Nuclear Information System (INIS)

Anderegg, F.; Behn, R.; Paris, P.J.; Salito, S.A.; Siegrist, M.R.; Weisen, H.

1988-06-01

There are several different and in general complementary methods for the investigation of plasmas. All of them have different characteristics and properties covering a large spectrum of physical measuring techniques. Electromagnetic waves serving as 'thermometers' permit to detect the global behaviour of the plasma as well as that of the particles composing it. One of the advantages of these introspective methods is that it brings information on temporary and local conditions of the domain being interrogated. With the development of micro-wave sources and lasers after the war the principal tools of this type of plasma diagnostics are now available. In this paper the emphasis is on the lasers which are different according to the type of measurement. Their versatility in measuring plasma parameters is largely acknowledged. We illustrate the potential of measuring methods by lasers by means of the research work done at two experimental installations of CRPP. (author) 21 figs., 8 refs

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

A Simple Model for Nonlinear Confocal Ultrasonic Beams

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Zhang, Dong; Zhou, Lin; Si, Li-Sheng; Gong, Xiu-Fen

2007-01-01

A confocally and coaxially arranged pair of focused transmitter and receiver represents one of the best geometries for medical ultrasonic imaging and non-invasive detection. We develop a simple theoretical model for describing the nonlinear propagation of a confocal ultrasonic beam in biological tissues. On the basis of the parabolic approximation and quasi-linear approximation, the nonlinear Khokhlov-Zabolotskaya-Kuznetsov (KZK) equation is solved by using the angular spectrum approach. Gaussian superposition technique is applied to simplify the solution, and an analytical solution for the second harmonics in the confocal ultrasonic beam is presented. Measurements are performed to examine the validity of the theoretical model. This model provides a preliminary model for acoustic nonlinear microscopy.

Texture-based characterization of subskin features by specified laser speckle effects at λ = 650 nm region for more accurate parametric 'skin age' modelling.

Science.gov (United States)

Orun, A B; Seker, H; Uslan, V; Goodyer, E; Smith, G

2017-06-01

The textural structure of 'skin age'-related subskin components enables us to identify and analyse their unique characteristics, thus making substantial progress towards establishing an accurate skin age model. This is achieved by a two-stage process. First by the application of textural analysis using laser speckle imaging, which is sensitive to textural effects within the λ = 650 nm spectral band region. In the second stage, a Bayesian inference method is used to select attributes from which a predictive model is built. This technique enables us to contrast different skin age models, such as the laser speckle effect against the more widely used normal light (LED) imaging method, whereby it is shown that our laser speckle-based technique yields better results. The method introduced here is non-invasive, low cost and capable of operating in real time; having the potential to compete against high-cost instrumentation such as confocal microscopy or similar imaging devices used for skin age identification purposes. © 2016 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

Confocal non-line-of-sight imaging based on the light-cone transform

Science.gov (United States)

O’Toole, Matthew; Lindell, David B.; Wetzstein, Gordon

2018-03-01

How to image objects that are hidden from a camera’s view is a problem of fundamental importance to many fields of research, with applications in robotic vision, defence, remote sensing, medical imaging and autonomous vehicles. Non-line-of-sight (NLOS) imaging at macroscopic scales has been demonstrated by scanning a visible surface with a pulsed laser and a time-resolved detector. Whereas light detection and ranging (LIDAR) systems use such measurements to recover the shape of visible objects from direct reflections, NLOS imaging reconstructs the shape and albedo of hidden objects from multiply scattered light. Despite recent advances, NLOS imaging has remained impractical owing to the prohibitive memory and processing requirements of existing reconstruction algorithms, and the extremely weak signal of multiply scattered light. Here we show that a confocal scanning procedure can address these challenges by facilitating the derivation of the light-cone transform to solve the NLOS reconstruction problem. This method requires much smaller computational and memory resources than previous reconstruction methods do and images hidden objects at unprecedented resolution. Confocal scanning also provides a sizeable increase in signal and range when imaging retroreflective objects. We quantify the resolution bounds of NLOS imaging, demonstrate its potential for real-time tracking and derive efficient algorithms that incorporate image priors and a physically accurate noise model. Additionally, we describe successful outdoor experiments of NLOS imaging under indirect sunlight.

In vivo confocal microscopy of conjunctiva-associated lymphoid tissue in healthy humans.

Science.gov (United States)

Agnifili, Luca; Mastropasqua, Rodolfo; Fasanella, Vincenzo; Di Staso, Silvio; Mastropasqua, Alessandra; Brescia, Lorenza; Mastropasqua, Leonardo

2014-07-29

To investigate modifications with aging of the presence, distribution and morphologic features of conjunctiva-associated lymphoid tissue (CALT) in healthy human subjects using laser scanning in vivo confocal microscopy (IVCM). A total of 108 (age range, 17-75 years) subjects were enrolled. In vivo confocal microscopy of the tarsal and bulbar conjunctiva, and impression cytology (IC) with CD3 (intra-epithelial T-lymphocytes) and CD20 (intra-epithelial B-lymphocytes) antibody immunofluorescence staining were performed. The main outcomes were subepithelial lymphocyte density (LyD), follicular density (FD), and follicular area (FA). The secondary outcomes were follicular reflectivity (FR), and lymphocyte density (FLyD), and CD3 and CD20 positivity. Conjunctiva-associated lymphoid tissue was observed in all subjects (97% only superior and 3% in both superior and inferior tarsum). Lymphocyte density ranged from 7.8 to 165.8 cells/mm(2) (46.42 [18.37]; mean [SD]), FD from 0.5 to 19.4 follicles/mm(2) (5.3 [3.6]), and FA from 1110 to 96,280 mm(2) (26,440 [26,280]). All three parameters showed a highly significant inverse cubic relationship with age (P lymphoid structures. These modifications may account for the decrease of mucosal immune response and increase of ocular surface diseases in the elderly. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

Mapping microscopic order in plant and mammalian cells and tissues: novel differential polarization attachment for new generation confocal microscopes (DP-LSM)

Science.gov (United States)

Steinbach, G.; Pawlak, K.; Pomozi, I.; Tóth, E. A.; Molnár, A.; Matkó, J.; Garab, G.

2014-03-01

Elucidation of the molecular architecture of complex, highly organized molecular macro-assemblies is an important, basic task for biology. Differential polarization (DP) measurements, such as linear (LD) and circular dichroism (CD) or the anisotropy of the fluorescence emission (r), which can be carried out in a dichrograph or spectrofluorimeter, respectively, carry unique, spatially averaged information about the molecular organization of the sample. For inhomogeneous samples—e.g. cells and tissues—measurements on macroscopic scale are not satisfactory, and in some cases not feasible, thus microscopic techniques must be applied. The microscopic DP-imaging technique, when based on confocal laser scanning microscope (LSM), allows the pixel by pixel mapping of anisotropy of a sample in 2D and 3D. The first DP-LSM configuration, which, in fluorescence mode, allowed confocal imaging of different DP quantities in real-time, without interfering with the ‘conventional’ imaging, was built on a Zeiss LSM410. It was demonstrated to be capable of determining non-confocally the linear birefringence (LB) or LD of a sample and, confocally, its FDLD (fluorescence detected LD), the degree of polarization (P) and the anisotropy of the fluorescence emission (r), following polarized and non-polarized excitation, respectively (Steinbach et al 2009 Acta Histochem.111 316-25). This DP-LSM configuration, however, cannot simply be adopted to new generation microscopes with considerably more compact structures. As shown here, for an Olympus FV500, we designed an easy-to-install DP attachment to determine LB, LD, FDLD and r, in new-generation confocal microscopes, which, in principle, can be complemented with a P-imaging unit, but specifically to the brand and type of LSM.

Mapping microscopic order in plant and mammalian cells and tissues: novel differential polarization attachment for new generation confocal microscopes (DP-LSM)

International Nuclear Information System (INIS)

Steinbach, G; Pawlak, K; Garab, G; Pomozi, I; Tóth, E A; Molnár, A; Matkó, J

2014-01-01

Elucidation of the molecular architecture of complex, highly organized molecular macro-assemblies is an important, basic task for biology. Differential polarization (DP) measurements, such as linear (LD) and circular dichroism (CD) or the anisotropy of the fluorescence emission (r), which can be carried out in a dichrograph or spectrofluorimeter, respectively, carry unique, spatially averaged information about the molecular organization of the sample. For inhomogeneous samples—e.g. cells and tissues—measurements on macroscopic scale are not satisfactory, and in some cases not feasible, thus microscopic techniques must be applied. The microscopic DP-imaging technique, when based on confocal laser scanning microscope (LSM), allows the pixel by pixel mapping of anisotropy of a sample in 2D and 3D. The first DP-LSM configuration, which, in fluorescence mode, allowed confocal imaging of different DP quantities in real-time, without interfering with the ‘conventional’ imaging, was built on a Zeiss LSM410. It was demonstrated to be capable of determining non-confocally the linear birefringence (LB) or LD of a sample and, confocally, its FDLD (fluorescence detected LD), the degree of polarization (P) and the anisotropy of the fluorescence emission (r), following polarized and non-polarized excitation, respectively (Steinbach et al 2009 Acta Histochem.111 316–25). This DP-LSM configuration, however, cannot simply be adopted to new generation microscopes with considerably more compact structures. As shown here, for an Olympus FV500, we designed an easy-to-install DP attachment to determine LB, LD, FDLD and r, in new-generation confocal microscopes, which, in principle, can be complemented with a P-imaging unit, but specifically to the brand and type of LSM. (paper)

Digital Position Encoding Of Galvanometer Scanner In A Laser Microscope

Science.gov (United States)

Liljeborg, Anders

1988-09-01

An account is given of a realization of a feedback method to digitize the analog position signal from a moving iron galvanometer. It is employed in a confocal scanning laser microscope for generating digital images. The photometric sampling has to be closely coupled to the position of a mirror that scans a focused laser beam across a microscope specimen. Pictures with low geometric distortion are obtained up to the size 1024 x 1024 pixels.

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.

Conduction cooled compact laser for the chemcam instrument

Science.gov (United States)

Durand, E.; Derycke, C.; Simon-Boisson, C.; Muller, S.; Faure, B.; Saccoccio, M.; Maurice, M.

2017-11-01

A new conduction cooled compact laser for laser induced spectroscopy on the Mars Science Laboratory (MSL) to be launched in 2009 is presented. An oscillator combined to amplifiers generates 30mJ at 1μm with a good spatial quality. Development prototype of this laser has been built and characterized. Environmental testing of this prototype is also reported.

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.

Combination of Small Molecule Microarray and Confocal Microscopy Techniques for Live Cell Staining Fluorescent Dye Discovery

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Attila Bokros

2013-08-01

Full Text Available Discovering new fluorochromes is significantly advanced by high-throughput screening (HTS methods. In the present study a combination of small molecule microarray (SMM prescreening and confocal laser scanning microscopy (CLSM was developed in order to discover novel cell staining fluorescent dyes. Compounds with high native fluorescence were selected from a 14,585-member library and further tested on living cells under the microscope. Eleven compartment-specific, cell-permeable (or plasma membrane-targeted fluorochromes were identified. Their cytotoxicity was tested and found that between 1–10 micromolar range, they were non-toxic even during long-term incubations.

Tokamak T-10 multipulse laser scattering: Instrumentation modernization

International Nuclear Information System (INIS)

Baukov, V.A.; Ponomarev, A.V.; Gorshkov, A.V.; Rossikhin, B.A.; Sannikov, V.V.; Grek, B.

1994-01-01

The modernized Thomson scattering diagnostic complex of Tokamak T-10 is described. The complex is based on a high-power neodimium laser. Both the laser fundamental and second harmonics serve to find out the time-dependence of plasma electron temperature and density profiles, as well as to measure the electron distribution function. One detected the scattered laser radiation at θ = 90 degrees and 2.5 degrees angles. The new-version Nd laser generates eight-pulse sequences. The laser radiation energy is E o = 30-50 J/pulse. The radiation divergence was smaller than var-phi = 0.15 mrad. The multiple radiation parameters were found to be very stable. The operator could vary the inter-pulse time intervals within the pulse sequence. The second-harmonic radiation energy was E 2 = 10-15 J/pulse. The data acquisition and analysis system was supported by IBM/AT and Macintosh computers. 6 refs., 1 fig

InGaP - CREATE portal | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available : Fixed mouse brain sections were fluorescent-labeled and observed by confocal laser scanning microscope usi...ted GFP-fusion KIAA/mKIAA in HEK293 cells were observed by fluorescent microscope...ope (neocortex 1) Observation by Confocal laser scanning microscope (neocortex 1) C...onfocal laser scanning microscope (neocortex 2) Observation by Confocal laser scanning microscope (neocortex... 2) Confocal laser scanning microscope (hippocampus) Observation by Confocal laser scanning microscope

A combined remote Raman and LIBS instrument for characterizing minerals with 532 nm laser excitation.

Science.gov (United States)

Sharma, Shiv K; Misra, Anupam K; Lucey, Paul G; Lentz, Rachel C F

2009-08-01

The authors have developed an integrated remote Raman and laser-induced breakdown spectroscopy (LIBS) system for measuring both the Raman and LIBS spectra of minerals with a single 532 nm laser line of 35 mJ/pulse and 20 Hz. The instrument has been used for analyzing both Raman and LIBS spectra of carbonates, sulfates, hydrous and anhydrous silicates, and iron oxide minerals in air. These experiments demonstrate that by focusing a frequency-doubled 532 nm Nd:YAG pulsed laser beam with a 10x beam expander to a 529-microm diameter spot on a mineral surface located at 9 m, it is possible to measure simultaneously both the remote Raman and LIBS spectra of calcite, gypsum and olivine by adjusting the laser power electronically. The spectra of calcite, gypsum, and olivine contain fingerprint Raman lines; however, it was not possible to measure the remote Raman spectra of magnetite and hematite at 9 m because of strong absorption of 532 nm laser radiation and low intensities of Raman lines from these minerals. The remote LIBS spectra of both magnetite and hematite contain common iron emission lines but show difference in the minor amount of Li present in these two minerals. Remote Raman and LIBS spectra of a number of carbonates, sulfates, feldspars and phyllosilicates at a distance of 9 m were measured with a 532-nm laser operating at 35 mJ/pulse and by changing photon flux density at the sample by varying the spot diameter from 10 mm for Raman to 530 microm for LIBS measurements. The complementary nature of these spectra is highlighted and discussed. The combined Raman and LIBS system can also be re-configured to perform micro-Raman and micro-LIBS analyses, which have applications in trace/residue analysis and analysis of very small samples in the nano-gram range.

Clinical applications of corneal confocal microscopy

Directory of Open Access Journals (Sweden)

Mitra Tavakoli

2008-06-01

Full Text Available Mitra Tavakoli1, Parwez Hossain2, Rayaz A Malik11Division of Cardiovascular Medicine, University of Manchester and Manchester Royal Infirmary, Manchester, UK; 2University of Southampton, Southampton Eye Unit, Southampton General Hospital, Southampton, UKAbstract: Corneal confocal microscopy is a novel clinical technique for the study of corneal cellular structure. It provides images which are comparable to in-vitro histochemical techniques delineating corneal epithelium, Bowman’s layer, stroma, Descemet’s membrane and the corneal endothelium. Because, corneal confocal microscopy is a non invasive technique for in vivo imaging of the living cornea it has huge clinical potential to investigate numerous corneal diseases. Thus far it has been used in the detection and management of pathologic and infectious conditions, corneal dystrophies and ecstasies, monitoring contact lens induced corneal changes and for pre and post surgical evaluation (PRK, LASIK and LASEK, flap evaluations and Radial Keratotomy, and penetrating keratoplasty. Most recently it has been used as a surrogate for peripheral nerve damage in a variety of peripheral neuropathies and may have potential in acting as a surrogate marker for endothelial abnormalities.Keywords: corneal confocal microscopy, cornea, infective keratitis, corneal dystrophy, neuropathy

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.

Two-color pump-probe laser spectroscopy instrument with picosecond time-resolved electronic delay and extended scan range

Science.gov (United States)

Yu, Anchi; Ye, Xiong; Ionascu, Dan; Cao, Wenxiang; Champion, Paul M.

2005-11-01

An electronically delayed two-color pump-probe instrument was developed using two synchronized laser systems. The instrument has picosecond time resolution and can perform scans over hundreds of nanoseconds without the beam divergence and walk-off effects that occur using standard spatial delay systems. A unique picosecond Ti :sapphire regenerative amplifier was also constructed without the need for pulse stretching and compressing optics. The picosecond regenerative amplifier has a broad wavelength tuning range, which suggests that it will make a significant contribution to two-color pump-probe experiments. To test this instrument we studied the rotational correlation relaxation of myoglobin (τr=8.2±0.5ns) in water as well as the geminate rebinding kinetics of oxygen to myoglobin (kg1=1.7×1011s-1, kg2=3.4×107s-1). The results are consistent with, and improve upon, previous studies.

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.

Imaging theory of nonlinear second harmonic and third harmonic generations in confocal microscopy

Institute of Scientific and Technical Information of China (English)

TANG Zhilie; XING Da; LIU Songhao

2004-01-01

The imaging theory of nonlinear second harmonic generation (SHG) and third harmonic generation (THG) in confocal microscopy is presented in this paper. The nonlinear effect of SHG and THG on the imaging properties of confocal microscopy has been analyzed in detail by the imaging theory. It is proved that the imaging process of SHG and THG in confocal microscopy, which is different from conventional coherent imaging or incoherent imaging, can be divided into two different processes of coherent imaging. The three-dimensional point spread functions (3D-PSF) of SHG and THG confocal microscopy are derived based on the nonlinear principles of SHG and THG. The imaging properties of SHG and THG confocal microscopy are discussed in detail according to its 3D-PSF. It is shown that the resolution of SHG and THG confocal microscopy is higher than that of single-and two-photon confocal microscopy.

Three dimensional subsurface elemental identification of minerals using confocal micro-X-ray fluorescence and micro-X-ray computed tomography

International Nuclear Information System (INIS)

Cordes, Nikolaus L.; Seshadri, Srivatsan; Havrilla, George J.; Yuan, Xiaoli; Feser, Michael; Patterson, Brian M.

2015-01-01

Current non-destructive elemental characterization methods, such as scanning electron microscopy-based energy dispersive spectroscopy (SEM–EDS) and micro-X-ray fluorescence spectroscopy (MXRF), are limited to either elemental identification at the surface (SEM–EDS) or suffer from an inability to discriminate between surface or depth information (MXRF). Thus, a non-destructive elemental characterization of individual embedded particles beneath the surface is impossible with either of these techniques. This limitation can be overcome by using laboratory-based 3D confocal micro-X-ray fluorescence spectroscopy (confocal MXRF). This technique utilizes focusing optics on the X-ray source and detector which allows for spatial discrimination in all three dimensions. However, the voxel-by-voxel serial acquisition of a 3D elemental scan can be very time-intensive (~ 1 to 4 weeks) if it is necessary to locate individual embedded particles of interest. As an example, if each point takes a 5 s measurement time, a small volume of 50 × 50 × 50 pixels leads to an acquisition time of approximately 174 h, not including sample stage movement time. Initially screening the samples for particles of interest using micro-X-ray computed tomography (micro-CT) can significantly reduce the time required to spatially locate these particles. Once located, these individual particles can be elementally characterized with confocal MXRF. Herein, we report the elemental identification of high atomic number surface and subsurface particles embedded in a mineralogical matrix by coupling micro-CT and confocal MXRF. Synergistically, these two X-ray based techniques first rapidly locate and then elementally identify individual subsurface particles. - Highlights: • Coupling of confocal X-ray fluorescence spectroscopy and X-ray computed tomography • Qualitative elemental identification of surface and subsurface mineral particles • Non-destructive particle size measurements • Utilization of

Three dimensional subsurface elemental identification of minerals using confocal micro-X-ray fluorescence and micro-X-ray computed tomography

Energy Technology Data Exchange (ETDEWEB)

Cordes, Nikolaus L., E-mail: ncordes@lanl.gov [Polymers and Coatings Group, Material Science and Technology Division, Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Seshadri, Srivatsan, E-mail: srivatsan.seshadri@zeiss.com [Carl Zeiss X-ray Microscopy, Inc., Pleasanton, CA 94588 (United States); Havrilla, George J. [Chemical Diagnostics and Engineering, Chemistry Division, Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Yuan, Xiaoli [Julius Kruttschnitt Mineral Research Centre, University of Queensland, Indooroopilly, Brisbane, QLD 4068 (Australia); Feser, Michael [Carl Zeiss X-ray Microscopy, Inc., Pleasanton, CA 94588 (United States); Patterson, Brian M. [Polymers and Coatings Group, Material Science and Technology Division, Los Alamos National Laboratory, Los Alamos, NM 87545 (United States)

2015-01-01

Current non-destructive elemental characterization methods, such as scanning electron microscopy-based energy dispersive spectroscopy (SEM–EDS) and micro-X-ray fluorescence spectroscopy (MXRF), are limited to either elemental identification at the surface (SEM–EDS) or suffer from an inability to discriminate between surface or depth information (MXRF). Thus, a non-destructive elemental characterization of individual embedded particles beneath the surface is impossible with either of these techniques. This limitation can be overcome by using laboratory-based 3D confocal micro-X-ray fluorescence spectroscopy (confocal MXRF). This technique utilizes focusing optics on the X-ray source and detector which allows for spatial discrimination in all three dimensions. However, the voxel-by-voxel serial acquisition of a 3D elemental scan can be very time-intensive (~ 1 to 4 weeks) if it is necessary to locate individual embedded particles of interest. As an example, if each point takes a 5 s measurement time, a small volume of 50 × 50 × 50 pixels leads to an acquisition time of approximately 174 h, not including sample stage movement time. Initially screening the samples for particles of interest using micro-X-ray computed tomography (micro-CT) can significantly reduce the time required to spatially locate these particles. Once located, these individual particles can be elementally characterized with confocal MXRF. Herein, we report the elemental identification of high atomic number surface and subsurface particles embedded in a mineralogical matrix by coupling micro-CT and confocal MXRF. Synergistically, these two X-ray based techniques first rapidly locate and then elementally identify individual subsurface particles. - Highlights: • Coupling of confocal X-ray fluorescence spectroscopy and X-ray computed tomography • Qualitative elemental identification of surface and subsurface mineral particles • Non-destructive particle size measurements • Utilization of

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

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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.

Measuring variations of δ18O and δ2H in atmospheric water vapour using two commercial laser-based spectrometers: an instrument characterisation study

Directory of Open Access Journals (Sweden)

S. Pfahl

2012-07-01

Full Text Available Variations of stable water isotopes in water vapour have become measurable at a measurement frequency of about 1 Hz in recent years using novel laser spectroscopic techniques. This enables us to perform continuous measurements for process-based investigations of the atmospheric water cycle at the time scales relevant for synoptic and mesoscale meteorology. An important prerequisite for the interpretation of data from automated field measurements lasting for several weeks or months is a detailed knowledge about instrument properties and the sources of measurement uncertainty. We present here a comprehensive characterisation and comparison study of two commercial laser spectroscopic systems based on cavity ring-down spectroscopy (Picarro and off-axis integrated cavity output spectroscopy (Los Gatos Research. The uncertainty components of the measurements were first assessed in laboratory experiments, focussing on the effects of (i water vapour mixing ratio, (ii measurement stability, (iii uncertainties due to calibration and (iv response times of the isotope measurements due to adsorption-desorption processes on the tubing and measurement cavity walls. Based on the experience from our laboratory experiments, we set up a one-week field campaign for comparing measurements of the ambient isotope signals from the two laser spectroscopic systems. The optimal calibration strategy determined for both instruments was applied as well as the correction functions for water vapour mixing ratio effects. The root mean square difference between the isotope signals from the two instruments during the field deployment was 2.3‰ for δ2H, 0.5‰ for δ18O and 3.1‰ for deuterium excess. These uncertainty estimates from field measurements compare well to those found in the laboratory experiments. The present quality of measurements from laser spectroscopic instruments combined with a calibration system opens new possibilities for investigating the atmospheric

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

A near-infrared confocal scanner

International Nuclear Information System (INIS)

Lee, Seungwoo; Yoo, Hongki

2014-01-01

In the semiconductor industry, manufacturing of three-dimensional (3D) packages or 3D integrated circuits is a high-performance technique that requires combining several functions in a small volume. Through-silicon vias, which are vertical electrical connections extending through a wafer, can be used to direct signals between stacked chips, thus increasing areal density by stacking and connecting multiple patterned chips. While defect detection is essential in the semiconductor manufacturing process, it is difficult to identify defects within a wafer or to monitor the bonding results between bonded surfaces because silicon and many other semiconductor materials are opaque to visible wavelengths. In this context, near-infrared (NIR) imaging is a promising non-destructive method to detect defects within silicon chips, to inspect bonding between chips and to monitor the chip alignment since NIR transmits through silicon. In addition, a confocal scanner provides high-contrast, optically-sectioned images of the specimen due to its ability to reject out-of-focus noise. In this study, we report an NIR confocal scanner that rapidly acquires high-resolution images with a large field of view through silicon. Two orthogonal line-scanning images can be acquired without rotating the system or the specimen by utilizing two orthogonally configured resonant scanning mirrors. This NIR confocal scanner can be efficiently used as an in-line inspection system when manufacturing semiconductor devices by rapidly detecting defects on and beneath the surface. (paper)

Model wavefront sensor for adaptive confocal microscopy

Science.gov (United States)

Booth, Martin J.; Neil, Mark A. A.; Wilson, Tony

2000-05-01

A confocal microscope permits 3D imaging of volume objects by the inclusion of a pinhole in the detector path which eliminates out of focus light. This configuration is however very sensitive to aberrations induced by the specimen or the optical system and would therefore benefit from an adaptive optics approach. We present a wavefront sensor capable of measuring directly the Zernike components of an aberrated wavefront and show that it is particularly applicable to the confocal microscope since only those wavefronts originating in the focal region contribute to the measured aberration.

A Spectroscopic Comparison of Femtosecond Laser Modified Fused Silica using kHz and MHz Laser Systems.

Energy Technology Data Exchange (ETDEWEB)

Reichman, W J; Krol, D M; Shah, L; Yoshino, F; Arai, A; Eaton, S M; Herman, P R

2005-09-29

Waveguides were written in fused silica using both a femtosecond fiber laser with a 1 MHz pulse repetition rate and a femtosecond amplified Ti:sapphire laser with a 1 kHz repetition rate. Confocal Raman and fluorescence microscopy were used to study structural changes in the waveguides written with both systems. A broad fluorescence band, centered at 650 nm, associated with non-bridging oxygen hole center (NBOHC) defects was observed after waveguide fabrication with the MHz laser. With the kHz laser system these defects were only observed for pulse energies above 1 {mu}J. Far fewer NBOHC defects were formed with the MHz laser than with kHz writing, possibly due to thermal annealing driven by heat accumulation effects at 1 MHz. When the kHz laser was used with pulse energies below 1 {mu}J, the predominant fluorescence was centered at 550 nm, a band assigned to the presence of silicon clusters (E{prime}{sub {delta}}). We also observed an increase in the intensity of the 605 cm{sup -1} Raman peak relative to the total Raman intensity, corresponding to an increase in the concentration of 3-membered rings in the lines fabricated with both laser systems.

Development of a fiber-coupled laser-induced breakdown spectroscopy instrument for analysis of underwater debris in a nuclear reactor core

International Nuclear Information System (INIS)

Saeki, Morihisa; Iwanade, Akio; Ohba, Hironori; Ito, Chikara; Wakaida, Ikuo; Thornton, Blair; Sakka, Tetsuo

2014-01-01

To inspect the post-accident nuclear core reactor of the TEPCO Fukushima Daiichi nuclear power plant (F1-NPP), a transportable fiber-coupled laser-induced breakdown spectroscopy (LIBS) instrument has been developed. The developed LIBS instrument was designed to analyze underwater samples in a high-radiation field by single-pulse breakdown with gas flow or double-pulse breakdown. To check the feasibility of the assembled fiber-coupled LIBS instrument for the analysis of debris material (mixture of the fuel core, fuel cladding, construction material and so on) in the F1-NPP, we investigated the influence of the radiation dose on the optical transmittance of the laser delivery fiber, compared data quality among various LIBS techniques for an underwater sample and studied the feasibility of the fiber-coupled LIBS system in an analysis of the underwater sample of the simulated debris in F1-NPP. In a feasible study conducted by using simulated debris, which was a mixture of CeO 2 (surrogate of UO 2 ), ZrO 2 and Fe, we selected atomic lines suitable for the analysis of materials, and prepared calibration curves for the component elements. The feasible study has guaranteed that the developed fiber-coupled LIBS system is applicable for analyzing the debris materials in the F1-NPP. (author)

Integration of instrumentation and processing software of a laser speckle contrast imaging system

Science.gov (United States)

Carrick, Jacob J.

Laser speckle contrast imaging (LSCI) has the potential to be a powerful tool in medicine, but more research in the field is required so it can be used properly. To help in the progression of Michigan Tech's research in the field, a graphical user interface (GUI) was designed in Matlab to control the instrumentation of the experiments as well as process the raw speckle images into contrast images while they are being acquired. The design of the system was successful and is currently being used by Michigan Tech's Biomedical Engineering department. This thesis describes the development of the LSCI GUI as well as offering a full introduction into the history, theory and applications of LSCI.

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.

A LabVIEW-Based Virtual Instrument System for Laser-Induced Fluorescence Spectroscopy.

Science.gov (United States)

Wu, Qijun; Wang, Lufei; Zu, Lily

2011-01-01

We report the design and operation of a Virtual Instrument (VI) system based on LabVIEW 2009 for laser-induced fluorescence experiments. This system achieves synchronous control of equipment and acquisition of real-time fluorescence data communicating with a single computer via GPIB, USB, RS232, and parallel ports. The reported VI system can also accomplish data display, saving, and analysis, and printing the results. The VI system performs sequences of operations automatically, and this system has been successfully applied to obtain the excitation and dispersion spectra of α-methylnaphthalene. The reported VI system opens up new possibilities for researchers and increases the efficiency and precision of experiments. The design and operation of the VI system are described in detail in this paper, and the advantages that this system can provide are highlighted.

Optical sectioning using a digital Fresnel incoherent-holography-based confocal imaging system

OpenAIRE

Kelner, Roy; Katz, Barak; Rosen, Joseph

2014-01-01

We propose a new type of confocal microscope using Fresnel incoherent correlation holography (FINCH). Presented here is a confocal configuration of FINCH using a phase pinhole and point illumination that is able to suppress out-of-focus information from the recorded hologram and hence combine the super-resolution capabilities of FINCH with the sectioning capabilities of confocal microscopy.

Evaluation and purchase of confocal microscopes: Numerous factors to consider

Science.gov (United States)

The purchase of a confocal microscope can be a complex and difficult decision for an individual scientist, group or evaluation committee. This is true even for scientists that have used confocal technology for many years. The task of reaching the optimal decision becomes almost i...

Laser sampling

International Nuclear Information System (INIS)

Gorbatenko, A A; Revina, E I

2015-01-01

The review is devoted to the major advances in laser sampling. The advantages and drawbacks of the technique are considered. Specific features of combinations of laser sampling with various instrumental analytical methods, primarily inductively coupled plasma mass spectrometry, are discussed. Examples of practical implementation of hybrid methods involving laser sampling as well as corresponding analytical characteristics are presented. The bibliography includes 78 references

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.

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)

Progress in the specification of optical instruments for the measurement of surface form and texture

Science.gov (United States)

de Groot, Peter J.

2014-05-01

Specifications for confocal microscopes, optical interferometers and other methods of measuring areal surface topography can be confusing and misleading. The emerging ISO 25178 standards, together with the established international vocabulary of metrology, provide a foundation for improved specifications for 3D surface metrology instrumentation. The approach in this paper links instrument specifications to metrological characteristics that can influence a measurement, using consistent definitions of terms, and reference to verification procedures.

Optical sectioning using a digital Fresnel incoherent-holography-based confocal imaging system

Science.gov (United States)

Kelner, Roy; Katz, Barak; Rosen, Joseph

2015-01-01

We propose a new type of confocal microscope using Fresnel incoherent correlation holography (FINCH). Presented here is a confocal configuration of FINCH using a phase pinhole and point illumination that is able to suppress out-of-focus information from the recorded hologram and hence combine the super-resolution capabilities of FINCH with the sectioning capabilities of confocal microscopy. PMID:26413560

Quantitative assessment of wound-healing process as a response to laser-induced micro-injuries

Science.gov (United States)

Liu, Yang; Bargo, Paulo; Kollias, Nikiforos

2009-02-01

Currently, most investigations of wound healing rely on invasive biopsy followed by histology and immunohistochemistry staining. There is a great need to develop non-invasive techniques for in vivo diagnostic, clinical and scientific evaluation. Here, we performed a comprehensive investigation on the dynamic wound healing process as a response to laser-induced microinjuries using non-invasive imaging techniques such as reflectance laser-scanning confocal microscopy and video microscopy. Eight healthy subjects ranging from Fitzpatrick skin type II-VI with age from 27 to 57 years were recruited. The volar forearm of each subject was treated with a laser device that generates an array of microbeams with an infrared wavelength. The microscopic changes of epidermal cells and collagen during the wound healing process were assessed non-invasively using confocal microscopy. We also developed a quantitative method to evaluate the dynamic wound healing process at the microscopic level in three areas of interest: (1) treated micro-wounding zone, (2) surrounding collateral damage zone and (3) normal area. The depth-dependent intensity profile derived from reflectance confocal microscope images clearly distinguishes the three areas of interest and quantitatively measures the cellular structure-associated changes. A progressive change in depth-dependent intensity profiles in subjects with different ages parallels the clinical observation of wound healing rate. The quantitative analysis developed in this study may find broad applications in assessing the skin response to treatment at a microscopic level.

Characterization of LiF-based soft X-ray imaging detectors by confocal fluorescence microscopy

International Nuclear Information System (INIS)

Bonfigli, F; Gaudio, P; Lupelli, I; Nichelatti, E; Richetta, M; Vincenti, M A; Montereali, R M

2010-01-01

X-ray microscopy represents a powerful tool to obtain images of samples with very high spatial resolution. The main limitation of this technique is represented by the poor spatial resolution of standard imaging detectors. We proposed an innovative high-performance X-ray imaging detector based on the visible photoluminescence of colour centres in lithium fluoride. In this work, a confocal microscope in fluorescence mode was used to characterize LiF-based imaging detectors measuring CC integrated visible fluorescence signals of LiF crystals and films (grown on several kinds of substrates) irradiated by soft X-rays produced by a laser plasma source in different exposure conditions. The results are compared with the CC photoluminescence spectra measured on the same samples and discussed.

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.

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.

Thermal Testing and Model Correlation for Advanced Topographic Laser Altimeter Instrument (ATLAS)

Science.gov (United States)

Patel, Deepak

2016-01-01

The Advanced Topographic Laser Altimeter System (ATLAS) part of the Ice Cloud and Land Elevation Satellite 2 (ICESat-2) is an upcoming Earth Science mission focusing on the effects of climate change. The flight instrument passed all environmental testing at GSFC (Goddard Space Flight Center) and is now ready to be shipped to the spacecraft vendor for integration and testing. This topic covers the analysis leading up to the test setup for ATLAS thermal testing as well as model correlation to flight predictions. Test setup analysis section will include areas where ATLAS could not meet flight like conditions and what were the limitations. Model correlation section will walk through changes that had to be made to the thermal model in order to match test results. The correlated model will then be integrated with spacecraft model for on-orbit predictions.

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.

Fabry-Perot confocal resonator optical associative memory

Science.gov (United States)

Burns, Thomas J.; Rogers, Steven K.; Vogel, George A.

1993-03-01

A unique optical associative memory architecture is presented that combines the optical processing environment of a Fabry-Perot confocal resonator with the dynamic storage and recall properties of volume holograms. The confocal resonator reduces the size and complexity of previous associative memory architectures by folding a large number of discrete optical components into an integrated, compact optical processing environment. Experimental results demonstrate the system is capable of recalling a complete object from memory when presented with partial information about the object. A Fourier optics model of the system's operation shows it implements a spatially continuous version of a discrete, binary Hopfield neural network associative memory.

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

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

A multimodal instrument for real-time in situ study of ultrasound and cavitation mediated drug delivery

OpenAIRE

Bian, S; Seth, A; Daly, D; Carlisle, R; Stride, E

2017-01-01

The development of a multimodal instrument capable of real-time in situ measurements of cavitation activity and effect in tissue mimicking phantoms during ultrasound and cavitation mediated drug delivery experiments is described here. The instrument features an acoustic arm that can expose phantoms to high-intensity focused-ultrasound while measuring cavitation activity and an optical arm that monitors cavitation effect using confocal microscopy. This combination of modalities allows real-tim...

Development and Preliminary Tests of an Open-Path Airborne Diode Laser Absorption Instrument for Carbon Dioxide

Science.gov (United States)

Diskin, Glenn S.; DiGangi, Joshua P.; Yang, Melissa; Slate, Thomas A.; Rana, Mario

2015-01-01

Carbon dioxide (CO2) is well known for its importance as an atmospheric greenhouse gas, with many sources and sinks around the globe. Understanding the fluxes of carbon into and out of the atmosphere is a complex and daunting challenge. One tool applied by scientists to measure the vertical flux of CO2 near the surface uses the eddy covariance technique, most often from towers but also from aircraft flying specific patterns over the study area. In this technique, variations of constituents of interest are correlated with fluctuations in the local vertical wind velocity. Measurement requirements are stringent, particularly with regard to precision, sensitivity to small changes, and temporal sampling rate. In addition, many aircraft have limited payload capability, so instrument size, weight, and power consumption are also important considerations. We report on the development and preliminary application of an airborne sensor for the measurement of atmospheric CO2. The instrument, modeled on the successful DLH (Diode Laser Hygrometer) series of instruments, has been tested in the laboratory and on the NASA DC-8 aircraft. Performance parameters such as accuracy, precision, sensitivity, specificity, and temporal response are discussed in the context of typical atmospheric variability and suitability for flux measurement applications. On-aircraft, in-flight data have been obtained and are discussed as well. Performance of the instrument has been promising, and continued flight testing is planned during 2016.

Research and application on imaging technology of line structure light based on confocal microscopy

Science.gov (United States)

Han, Wenfeng; Xiao, Zexin; Wang, Xiaofen

2009-11-01

In 2005, the theory of line structure light confocal microscopy was put forward firstly in China by Xingyu Gao and Zexin Xiao in the Institute of Opt-mechatronics of Guilin University of Electronic Technology. Though the lateral resolution of line confocal microscopy can only reach or approach the level of the traditional dot confocal microscopy. But compared with traditional dot confocal microscopy, it has two advantages: first, by substituting line scanning for dot scanning, plane imaging only performs one-dimensional scanning, with imaging velocity greatly improved and scanning mechanism simplified, second, transfer quantity of light is greatly improved by substituting detection hairline for detection pinhole, and low illumination CCD is used directly to collect images instead of photoelectric intensifier. In order to apply the line confocal microscopy to practical system, based on the further research on the theory of the line confocal microscopy, imaging technology of line structure light is put forward on condition of implementation of confocal microscopy. Its validity and reliability are also verified by experiments.

Naural Responses to Injury: Prevention, Protection, and Repair. Volume 8. Vision, Laser Eye Injury, and Infectious Diseases

National Research Council Canada - National Science Library

Bazan, Nicolas

1997-01-01

Four specific aims were proposed in the original grant application. Develop a new confocal microscope that can be used in living eyes to understand the earliest stages of trauma, laser injuries, and disease...

Detecting plant silica fibres in animal tissue by confocal fluorescence microscopy.

Science.gov (United States)

Hodson, M J; Smith, R J; van Blaaderen, A; Crafton, T; O'Neill, C H

1994-04-01

Silica fibres from the inflorescence bracts of the grass Phalaris canariensis L. cause dermatitis, and have been implicated in the aetiology of oesophageal cancer in northeastern Iran. Here we describe a method for labelling these fibres so that they can be located in mammalian tissue. Fluorescein was covalently linked to isolated, purified fibres with the silane coupling agent 3-aminopropyl triethoxysilane. The labelled hairs were then rubbed into the backs of mice. These were later killed and their skin fixed, stained and sliced at a thickness of 250 microns. A confocal laser scanning microscope gave brilliant images of the fibres at any depth up to 100 microns or more beneath the surface of the slice. Fibres penetrated deeply into the dermis. Several cubic millimetres of tissue could be surveyed in 1 h. The number of fibres present was approximately 2 mm-3 initially, falling to 0.1 mm-3 after 7 days.

Laser-based instrumentation for the detection of chemical agents

International Nuclear Information System (INIS)

Hartford, A. Jr.; Sander, R.K.; Quigley, G.P.; Radziemski, L.J.; Cremers, D.A.

1982-01-01

Several laser-based techniques are being evaluated for the remote, point, and surface detection of chemical agents. Among the methods under investigation are optoacoustic spectroscopy, laser-induced breakdown spectroscopy (LIBS), and synchronous detection of laser-induced fluorescence (SDLIF). Optoacoustic detection has already been shown to be capable of extremely sensitive point detection. Its application to remote sensing of chemical agents is currently being evaluated. Atomic emission from the region of a laser-generated plasma has been used to identify the characteristic elements contained in nerve (P and F) and blister (S and Cl) agents. Employing this LIBS approach, detection of chemical agent simulants dispersed in air and adsorbed on a variety of surfaces has been achieved. Synchronous detection of laser-induced fluorescence provides an attractive alternative to conventional LIF, in that an artificial narrowing of the fluorescence emission is obtained. The application of this technique to chemical agent simulants has been successfully demonstrated. 19 figures

Nerve fiber layer (NFL) degeneration associated with acute q-switched laser exposure in the nonhuman primate

Science.gov (United States)

Zwick, Harry; Zuclich, Joseph A.; Stuck, Bruce E.; Gagliano, Donald A.; Lund, David J.; Glickman, Randolph D.

1995-01-01

We have evaluated acute laser retinal exposure in non-human primates using a Rodenstock scanning laser ophthalmoscope (SLO) equipped with spectral imaging laser sources at 488, 514, 633, and 780 nm. Confocal spectral imaging at each laser wavelength allowed evaluation of the image plane from deep within the retinal vascular layer to the more superficial nerve fiber layer in the presence and absence of the short wavelength absorption of the macular pigment. SLO angiography included both fluorescein and indocyanine green procedures to assess the extent of damage to the sensory retina, the retinal pigment epithelium (RPE), and the choroidal vasculature. All laser exposures in this experiment were from a Q-switched Neodymium laser source at an exposure level sufficient to produce vitreous hemorrhage. Confocal imaging of the nerve fiber layer revealed discrete optic nerve sector defects between the lesion site and the macula (retrograde degeneration) as well as between the lesion site and the optic disk (Wallerian degeneration). In multiple hemorrhagic exposures, lesions placed progressively distant from the macula or overlapping the macula formed bridging scars visible at deep retinal levels. Angiography revealed blood flow disturbance at the retina as well as at the choroidal vascular level. These data suggest that acute parafoveal laser retinal injury can involve both direct full thickness damage to the sensory and non-sensory retina and remote nerve fiber degeneration. Such injury has serious functional implications for both central and peripheral visual function.

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.

Numerical study of a confocal ultrasonic setup for creation of cavitation

Energy Technology Data Exchange (ETDEWEB)

Lafond, Maxime, E-mail: maxime.lafond@inserm.fr; Chavrier, Françoise; Prieur, Fabrice [Inserm, U1032, LabTau, Lyon, F-69003 (France); Université de Lyon, Lyon, F-69003 (France); Université Lyon 1, Lyon, F-69003 (France); Mestas, Jean-Louis; Lafon, Cyril [Inserm, U1032, LabTau, Lyon, F-69003 (France); Université de Lyon, Lyon, F-69003 (France); Université Lyon 1, Lyon, F-69003 (France); Caviskills SAS, Vaulx-En-Velin, F-69120 (France)

2015-10-28

Acoustic cavitation is used for various therapeutic applications such as local enhancement of drug delivery, histotripsy or hyperthermia. One of the utmost important parameter for cavitation creation is the rarefaction pressure. The typical magnitude of the rarefaction pressure required to initiate cavitation from gas dissolved in tissue is beyond the range of the megapascal. Because nonlinear effects need to be taken into account, a numerical simulator based on the Westervelt equation was used to study the pressure waveform and the acoustic field generated by a setup for creation of cavitation consisting of two high intensity focused ultrasound transducers mounted confocally. At constant acoustic power, simulations with only one and both transducers from the confocal setup showed that the distortion of the pressure waveform due to the combined effects of nonlinearity and diffraction is less pronounced when both confocal transducers are used. Consequently, the confocal setup generates a greater peak negative pressure at focus which is more favorable for cavitation initiation. Comparison between the confocal setup and a single transducer with the same total emitting surface puts in evidence the role of the spatial separation of the two beams. Furthermore, it has been previously shown that the location of the peak negative pressure created by a single transducer shifts from focus towards the transducers in the presence of nonlinear effects. The simulator was used to study a configuration where the acoustical axes of transducers intersect on the peak negative pressure instead of the geometrical focus. For a representative confocal setup, namely moderate nonlinear effects, a 2% increase of the peak negative pressure and 8% decrease of the peak positive pressure resulted from this configuration. These differences tend to increase by increasing nonlinear effects. Although the optimal position of the transducers varies with the nonlinear regimen, the intersection point

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.

Magnetically Triggered Release From Giant Unilamellar Vesicles: Visualization By Means Of Confocal Microscopy

KAUST Repository

Nappini, Silvia

2011-04-07

Magnetically triggered release from magnetic giant unilamellar vesicles (GUVs) loaded with Alexa fluorescent dye was studied by means of confocal laser scanning microscopy (CLSM) under a low-frequency alternating magnetic field (LF-AMF). Core/shell cobalt ferrite nanoparticles coated with rhodamine B isothiocyanate (MP@SiO 2(RITC)) were prepared and adsorbed on the GUV membrane. The MP@SiO 2(RITC) location and distribution on giant lipid vesicles were determined by 3D-CLSM projections, and their effect on the release properties and GUV permeability under a LF-AMF was investigated by CLSM time-resolved experiments. We show that the mechanism of release of the fluorescent dye during the LF-AMF exposure is induced by magnetic nanoparticle energy and mechanical vibration, which promote the perturbation of the GUV membrane without its collapse. © 2011 American Chemical Society.

Combination of confocal principle and aperture stop separation improves suppression of crystalline lens fluorescence in an eye model.

Science.gov (United States)

Klemm, Matthias; Blum, Johannes; Link, Dietmar; Hammer, Martin; Haueisen, Jens; Schweitzer, Dietrich

2016-09-01

Fluorescence lifetime imaging ophthalmoscopy (FLIO) is a new technique to detect changes in the human retina. The autofluorescence decay over time, generated by endogenous fluorophores, is measured in vivo. The strong autofluorescence of the crystalline lens, however, superimposes the intensity decay of the retina fluorescence, as the confocal principle is not able to suppress it sufficiently. Thus, the crystalline lens autofluorescence causes artifacts in the retinal fluorescence lifetimes determined from the intensity decays. Here, we present a new technique to suppress the autofluorescence of the crystalline lens by introducing an annular stop into the detection light path, which we call Schweitzer's principle. The efficacy of annular stops with an outer diameter of 7 mm and inner diameters of 1 to 5 mm are analyzed in an experimental setup using a model eye based on fluorescent dyes. Compared to the confocal principle, Schweitzer's principle with an inner diameter of 3 mm is able to reduce the simulated crystalline lens fluorescence to 4%, while 42% of the simulated retina fluorescence is preserved. Thus, we recommend the implementation of Schweitzer's principle in scanning laser ophthalmoscopes used for fundus autofluorescence measurements, especially the FLIO device, for improved image quality.

Proceedings of the twenty fourth DAE-BRNS national laser symposium: abstract book

International Nuclear Information System (INIS)

2015-01-01

The topics covered in this symposium are: physics and technology of lasers, lasers in nuclear science and technology, laser materials, devices and components, nonlinear, quantum optics and atomic optics, ultrafast lasers and applications, lasers in material science, laser plasma interaction, lasers in industry and defence, lasers in spectroscopy and applications, lasers in chemistry, biology and medicine, laser based instrumentation and electronics and instrumentation for lasers. Papers relevant to INIS are indexed separately

Quantification of Multilayer Samples by Confocal μXRF

International Nuclear Information System (INIS)

Perez, R. Daniel; Sanchez, H. J.; Rubio, M.; Perez, C. A.

2009-01-01

The confocal setup consists of x-ray lenses in the excitation as well as in the detection channel. In this configuration, a micro volume defined by the overlap of the foci of both x-ray lenses is analyzed. Scanning this micro volume through the sample, 1-3 dimensional studies can be performed. For intermediate thin homogeneous layers a scanning in the normal direction to the surface sample provides information of its thickness and elemental composition. For multilayer samples it also provides the order of each layer in the stratified structure. For the confocal setup, we used a glass monocapillary in the excitation channel and a monolithic half polycapillary in the detection channel. The experiment was carried out at the D09B beamline of the LNLS using white beam. In the present work, a new algorithm was applied to analyze in detail by confocal μXRF a sample of three paint layers on a glass substrate. Using the proposed algorithm, information about thickness and elemental densities was obtained for each layer of these samples.

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.

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.

Three-Dimensional Visualization of Interfacial Phenomena Using Confocal Microscopy

Science.gov (United States)

Shieh, Ian C.

Surfactants play an integral role in numerous functions ranging from stabilizing the emulsion in a favorite salad dressing to organizing the cellular components that make life possible. We are interested in lung surfactant, which is a mixture of lipids and proteins essential for normal respiration because it modulates the surface tension of the air-liquid interface of the thin fluid lining in the lungs. Through this surface tension modulation, lung surfactant ensures effortless lung expansion and prevents lung collapse during exhalation, thereby effecting proper oxygenation of the bloodstream. The function of lung surfactant, as well as numerous interfacial lipid systems, is not solely dictated by the behavior of materials confined to the two-dimensional interface. Rather, the distributions of materials in the liquid subphase also greatly influence the performance of interfacial films of lung surfactant. Therefore, to better understand the behavior of lung surfactant and other interfacial lipid systems, we require a three-dimensional characterization technique. In this dissertation, we have developed a novel confocal microscopy methodology for investigating the interfacial phenomena of surfactants at the air-liquid interface of a Langmuir trough. Confocal microscopy provides the excellent combination of in situ, fast, three-dimensional visualization of multiple components of the lung surfactant system that other characterization techniques lack. We detail the solutions to the numerous challenges encountered when imaging a dynamic air-liquid interface with a high-resolution technique like confocal microscopy. We then use confocal microscopy to elucidate the distinct mechanisms by which a polyelectrolyte (chitosan) and nonadsorbing polymer (polyethylene glycol) restore the function of lung surfactant under inhibitory conditions mimicking the effects of lung trauma. Beyond this physiological model, we also investigate several one- and two-component interfacial films

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.

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.

Multi-pollutants sensors based on near-IR telecom lasers and mid-IR difference frequency generation: development and applications; Instruments de mesure multi-polluants par spectroscopie infrarouge bases sur des lasers fibres et par generation de difference de frequences: developpement et applications

Energy Technology Data Exchange (ETDEWEB)

Cousin, J

2006-12-15

At present the detection of VOC and other anthropic trace pollutants is an important challenge in the measurement of air quality. Infrared spectroscopy, allowing spectral regions rich in molecular absorption to be probed, is a suitable technique for in-situ monitoring of the air pollution. Thus the aim of this work was to develop instruments capable of detecting multiple pollutants for in-situ monitoring by IR spectroscopy. A first project benefited from the availability of the telecommunications lasers emitting in near-IR. This instrument was based on an external cavity diode laser (1500 - 1640 nm) in conjunction with a multipass cell (100 m). The detection sensitivity was optimised by employing a balanced detection and a sweep integration procedure. The instrument developed is deployable for in-situ measurements with a sensitivity of < 10{sup -8} cm{sup -1} Hz{sup -1/2} and allowed the quantification of chemical species such as CO{sub 2}, CO, C{sub 2}H{sub 2}, CH{sub 4} and the determination of the isotopic ratio {sup 13}CO{sub 2}/{sup 12}CO{sub 2} in combustion environment The second project consisted in mixing two near-IR fiber lasers in a non-linear crystal (PPLN) in order to produce a laser radiation by difference frequency generation in the middle-IR (3.15 - 3.43 {mu}m), where the absorption bands of the molecules are the most intense. The first studies with this source were carried out on detection of ethylene (C{sub 2}H{sub 4}) and benzene (C{sub 6}H{sub 6}). Developments, characterizations and applications of these instruments in the near and middle IR are detailed and the advantages of the 2 spectral ranges is highlighted. (author)

Harvard ER-2 OH laser-induced fluorescence instrument

Science.gov (United States)

Wennberg, Paul O.; Anderson, James G.

1994-01-01

The Harvard ER-2 OH instrument is scheduled to be integrated into the NASA ER-2 high altitude aircraft ozone payload in August 1992. Design and fabrication is presently underway. This experiment is a descendant of a balloon borne instrument designed and built in the mid-1980s. The ER-2 instrument is being designed to measure OH and HO2 as part of the NASA ozone payload for the investigation of processes controlling the concentration of stratospheric ozone. Although not specifically designed to do so, it is hoped that valid measurements of OH and HO2 can be made in the remote free troposphere with this instrument.

X-ray laser studies using plasmas created by optical field ionization

International Nuclear Information System (INIS)

Krushelnick, K.M.; Tighe, W.; Suckewer, S.

1995-01-01

X-ray laser experiments involving the creation of fast recombining plasmas by optical field ionization of preformed targets were conducted. A nonlinear increase in the intensity of the 13.5nm Lyman-α line in Li III with the length of the target plasma was observed but only for distances less than the laser confocal parameter and for low plasma electron temperatures. Multiphoton pumping of resonant atomic transitions was also examined and the process of multiphoton ionization of FIII was found to be more probable than multiphoton excitation

Confocal microscopy patterns in nonmelanoma skin cancer and clinical applications.

Science.gov (United States)

González, S; Sánchez, V; González-Rodríguez, A; Parrado, C; Ullrich, M

2014-06-01

Reflectance confocal microscopy is currently the most promising noninvasive diagnostic tool for studying cutaneous structures between the stratum corneum and the superficial reticular dermis. This tool gives real-time images parallel to the skin surface; the microscopic resolution is similar to that of conventional histology. Numerous studies have identified the main confocal features of various inflammatory skin diseases and tumors, demonstrating the good correlation of these features with certain dermatoscopic patterns and histologic findings. Confocal patterns and diagnostic algorithms have been shown to have high sensitivity and specificity in melanoma and nonmelanoma skin cancer. Possible present and future applications of this noninvasive technology are wide ranging and reach beyond its use in noninvasive diagnosis. This tool can also be used, for example, to evaluate dynamic skin processes that occur after UV exposure or to assess tumor response to noninvasive treatments such as photodynamic therapy. We explain the characteristic confocal features found in the main nonmelanoma skin tumors and discuss possible applications for this novel diagnostic technique in routine dermatology practice. Copyright © 2012 Elsevier España, S.L. and AEDV. All rights reserved.

Laser polishing of additive manufactured Ti alloys

Science.gov (United States)

Ma, C. P.; Guan, Y. C.; Zhou, W.

2017-06-01

Laser-based additive manufacturing has attracted much attention as a promising 3D printing method for metallic components in recent years. However, surface roughness of additive manufactured components has been considered as a challenge to achieve high performance. In this work, we demonstrate the capability of fiber laser in polishing rough surface of additive manufactured Ti-based alloys as Ti-6Al-4V and TC11. Both as-received surface and laser-polished surfaces as well as cross-section subsurfaces were analyzed carefully by White-Light Interference, Confocal Microscope, Focus Ion Beam, Scanning Electron Microscopy, Energy Dispersive Spectrometer, and X-ray Diffraction. Results revealed that as-received Ti-based alloys with surface roughness more than 5 μm could be reduce to less than 1 μm through laser polishing process. Moreover, microstructure, microhardness and wear resistance of laser-polished zone was investigated in order to examine the thermal effect of laser polishing processing on the substrate of additive manufactured Ti alloys. This proof-of-concept process has the potential to effectively improve the surface roughness of additive manufactured metallic alloy by local polishing method without damage to the substrate.

Three-dimensional measurement and visualization of internal flow of a moving droplet using confocal micro-PIV.

Science.gov (United States)

Kinoshita, Haruyuki; Kaneda, Shohei; Fujii, Teruo; Oshima, Marie

2007-03-01

This paper presents a micro-flow diagnostic technique, 'high-speed confocal micro-particle image velocimetry (PIV)', and its application to the internal flow measurement of a droplet passing through a microchannel. A confocal micro-PIV system has been successfully constructed wherein a high-speed confocal scanner is combined with the conventional micro-PIV technique. The confocal micro-PIV system enables us to obtain a sequence of sharp and high-contrast cross-sectional particle images at 2000 frames s(-1). This study investigates the confocal depth, which is a significant parameter to determine the out-of-plane measurement resolution in confocal micro-PIV. Using the present confocal micro-PIV system, we can measure velocity distributions of micro-flows in a 228 microm x 171 microm region with a confocal depth of 1.88 microm. We also propose a three-dimensional velocity measurement method based on the confocal micro-PIV and the equation of continuity. This method enables us to measure three velocity components in a three-dimensional domain of micro flows. The confocal micro-PIV system is applied to the internal flow measurement of a droplet. We have measured three-dimensional distributions of three-component velocities of a droplet traveling in a 100 microm (width) x 58 microm (depth) channel. A volumetric velocity distribution inside a droplet is obtained by the confocal micro-PIV and the three-dimensional flow structure inside the droplet is investigated. The measurement results suggest that a three-dimensional and complex circulating flow is formed inside the droplet.

Laser Thomson Scattering, Raman Scattering and laser-absorption diagnostics of high pressure microdischarges

International Nuclear Information System (INIS)

Donnelly, Vincent M; Belostotskiy, Sergey G; Economou, Demetre J; Sadeghi, Nader

2010-01-01

Laser scattering experiments were performed in high pressure (100s of Torr) parallel-plate, slot-type DC microdischarges operating in argon or nitrogen. Laser Thomson Scattering (LTS) and Rotational Raman Scattering were employed in a novel, backscattering, confocal configuration. LTS allows direct and simultaneous measurement of both electron density (n e ) and electron temperature (T e ). For 50 mA current and over the pressure range of 300 - 700 Torr, LTS yielded T e = 0.9 ± 0.3 eV and n e = (6 ± 3)·10 13 cm -3 , in reasonable agreement with the predictions of a mathematical model. Rotational Raman spectroscopy (RRS) was employed for absolute calibration of the LTS signal. RRS was also applied to measure the 3D gas temperature (T g ) in nitrogen DC microdischarges. In addition, diode laser absorption spectroscopy was employed to measure the density of argon metastables (1s5 in Paschen notations) in argon microdischarges. The gas temperature, extracted from the width of the absorption profile, was compared with T g values obtained by optical emission spectroscopy.

Confocal microscopy for astrocyte in vivo imaging: Recycle and reuse in microscopy

Science.gov (United States)

Pérez-Alvarez, Alberto; Araque, Alfonso; Martín, Eduardo D.

2013-01-01

In vivo imaging is one of the ultimate and fundamental approaches for the study of the brain. Two-photon laser scanning microscopy (2PLSM) constitutes the state-of-the-art technique in current neuroscience to address questions regarding brain cell structure, development and function, blood flow regulation and metabolism. This technique evolved from laser scanning confocal microscopy (LSCM), which impacted the field with a major improvement in image resolution of live tissues in the 1980s compared to widefield microscopy. While nowadays some of the unparalleled features of 2PLSM make it the tool of choice for brain studies in vivo, such as the possibility to image deep within a tissue, LSCM can still be useful in this matter. Here we discuss the validity and limitations of LSCM and provide a guide to perform high-resolution in vivo imaging of the brain of live rodents with minimal mechanical disruption employing LSCM. We describe the surgical procedure and experimental setup that allowed us to record intracellular calcium variations in astrocytes evoked by sensory stimulation, and to monitor intact neuronal dendritic spines and astrocytic processes as well as blood vessel dynamics. Therefore, in spite of certain limitations that need to be carefully considered, LSCM constitutes a useful, convenient, and affordable tool for brain studies in vivo. PMID:23658537

Reflectance Confocal Microscopy in Lentigo Maligna.

Science.gov (United States)

Gamo, R; Pampín, A; Floristán, U

2016-12-01

Lentigo maligna is the most common type of facial melanoma. Diagnosis is complicated, however, as it shares clinical and dermoscopic characteristics with other cutaneous lesions of the face. Reflectance confocal microscopy is an imaging technique that permits the visualization of characteristic features of lentigo maligna. These include a disrupted honeycomb pattern and pagetoid cells with a tendency to show folliculotropism. These cells typically have a dendritic morphology, although they may also appear as round cells measuring over 20μm with atypical nuclei. Poorly defined dermal papillae and atypical cells may be seen at the dermal-epidermal junction and can form bridges resembling mitochondrial structures. Other characteristic findings include junctional swelling with atypical cells located around the follicles, resembling caput medusae. Reflectance confocal microscopy is a very useful tool for diagnosing lentigo maligna. Copyright © 2016 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.

Intravital Confocal and Two-photon Imaging of Dual-color Cells and Extracellular Matrix Mimics

Science.gov (United States)

Bal, Ufuk; Andresen, Volker; Baggett, Brenda; Utzinger, Urs

2013-01-01

To optimize imaging of cells in three dimensional culture we studied confocal backscattering, Second Harmonic Generation (SHG) and autofluorescence as source of contrast in extracellular matrix (ECM) mimics and evaluated the attenuation as well as bleaching of endogenous cellular fluorescence signals. All common ECM mimics exhibit contrast observable with confocal reflectance microscopy. SHG imaging on collagen I based hydrogels provides high contrast and good optical penetration depth. Agarose is a useful embedding medium because it allows for large optical penetration and exhibits minimal autofluorescence while still providing good reflectance to detect voids in the embedding medium. We labeled breast cancer cells’ outline with DsRed2 and nucleus with eGFP. DsRed2 can be excited with confocal imaging at 568nm, and with two photon excitation (TPE) in the red and longer NIR. eGFP was excited at 488nm for confocal and in the NIR for TPE. While there is small difference in the bleaching rate for eGFP between confocal and TPE we observed significant difference for DsRed2 where bleaching is strongest during TPE in the red wavelengths and smallest during confocal imaging. After a few hundred microns depth in a collagen I hydrogel, TPE fluorescence becomes twice as strong compared to confocal imaging. PMID:23380006

Comparison of laser-based mitigation of fused silica surface damage using mid- versus far-infrared lasers

Energy Technology Data Exchange (ETDEWEB)

Yang, S T; Matthews, M J; Elhadj, S; Cooke, D; Guss, G M; Draggoo, V G; Wegner, P J

2009-12-16

Laser induced growth of optical damage can limit component lifetime and therefore operating costs of large-aperture fusion-class laser systems. While far-infrared (IR) lasers have been used previously to treat laser damage on fused silica optics and render it benign, little is known about the effectiveness of less-absorbing mid-IR lasers for this purpose. In this study, they quantitatively compare the effectiveness and efficiency of mid-IR (4.6 {micro}m) versus far-IR (10.6 {micro}m) lasers in mitigating damage growth on fused silica surfaces. The non-linear volumetric heating due to mid-IR laser absorption is analyzed by solving the heat equation numerically, taking into account the temperature-dependent absorption coefficient {alpha}(T) at {lambda} = 4.6 {micro}m, while far-IR laser heating is well-described by a linear analytic approximation to the laser-driven temperature rise. In both cases, the predicted results agree well with surface temperature measurements based on infrared radiometry, as well as sub-surface fictive temperature measurements based on confocal Raman microscopy. Damage mitigation efficiency is assessed using a figure of merit (FOM) relating the crack healing depth to laser power required, under minimally-ablative conditions. Based on their FOM, they show that for cracks up to at least 500 {micro}m in depth, mitigation with a 4.6 {micro}m mid-IR laser is more efficient than mitigation with a 10.6 {micro}m far-IR laser. This conclusion is corroborated by direct application of each laser system to the mitigation of pulsed laser-induced damage possessing fractures up to 225 {micro}m in depth.

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

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

'In vitro' assessment to instrumented indentation hardness tests in enamel of bovine teeth, before and after dental bleaching by laser

International Nuclear Information System (INIS)

Britto Junior, Francisco Meira

2004-01-01

The laser enamel bleaching is a common used procedure due to its satisfactory esthetic results. The possible changes on the dental structures caused by the bleaching technique are of great importance. The enamel superficial microhardness changes through instrumented indentation hardness on bovine teeth were analyzed in this present study. The samples were divided in two halves, one being the control and the other irradiated with a diode laser (808 nm) or with a Nd:YAG laser (1064 nm) to activate the Whiteness HP bleaching gel (hydrogen peroxide at 35%). It was possible to conclude that there was a statistical significant increase on the enamel superficial microhardness (Group I, sample 1 and Group II, sample 1) despite this increase did not seem to indicate a concern regarding the enamel surface resistance change. There was not a significant statistical change on the enamel microhardness on the other samples. The final conclusion is that there was no superficial enamel morphological change after these treatments. (author)

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.

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

Lasers in oral surgery and implantology

Science.gov (United States)

Vescovi, Paolo

2016-03-01

The usefulness of laser for oral hard tissue procedure such as caries treatment, impacted teeth extraction, periodontal therapy, peri-implantitis management, sinus lifting is reported by several Authors [1]. Conventionally, mechanical rotary instruments and hand instruments are employed for bone surgery. Rotary instruments have better accessibility and cutting efficiency, but there is a risk of excessive heating of bone tissue and caution must be exercised to avoid the bur becoming entangled with surrounding soft tissues and the reflected flap. The main clinical advantages of the lasers are represented by minimal patient discomfort, good recovery with decreased or absent post-operative pain. In the last ten years are described in the international literature great advantages of Laser Surgery and Low Level Laser Therapy (LLLT) performed with different wavelength in addition to traditional surgical techniques to improve bone and soft tissue healing and for pain and infection control.

Experimental laser fusion devices and related vacuum problems

International Nuclear Information System (INIS)

O'Neal, W.C.; Campbell, D.E.; Glaros, S.S.; Hurley, C.A.; Kobierecki, M.W.; McFann, C.B. Jr.; Monjes, J.A.; Patton, H.G.; Rienecker, F. Jr.

1977-01-01

Laser fusion experiments require hard vacuum in the laser-beam spatial filters, target chambers and for target diagnostics instruments. Laser focusing lenses and windows, and target alignment windows must hold vacuum without optical distortion, and must be protected from target debris. The vacuum must be sufficient to prevent residual gas breakdown in focused laser light, avoid arcing at high voltage terminals, minimize contamination and melting of cryogenic targets, and prevent adsorption of the target's microfusion radiation before it reaches the diagnostics instruments

Confocal fluorescence microscopy investigation of visible emitting defects induced by electron beam lithography in LIF films

International Nuclear Information System (INIS)

Montereali, R. M.; Bigotta, S.; Pace, A.; Piccinini, M.; Burattini, E.; Grilli, A.; Raco, A.; Giammatteo, M.; L'Aquila Univ., L'Aquila; Picozzi, P.; Santucci, S.; L'Aquila Univ., L'Aquila

2000-01-01

Low energy electron irradiation of lithium fluoride (LiF), in the form of bulk crystals and films, gives rise to the stable formation of primary F defects and aggregated color centers in a thin layer located at the surface of the investigated material. For the first time a confocal light scanning microscope (CLSM) in fluorescence mode was used to reconstruct the depth distribution of efficiently emitting laser active color centers in a stripe-like region induced by 12 and 16 keV electrons on LiF films thermally evaporated on glass. The formation of the F3+ and F2 aggregated defects appears restricted to the electron penetration and proportional to their energy depth profile, as obtained from Monte Carlo simulations [it

The application of confocal technology based on polycapillary X-ray optics in surface topography

International Nuclear Information System (INIS)

Zhao, Guangcui; Sun, Tianxi; Liu, Zhiguo; Yuan, Hao; Li, Yude; Liu, Hehe; Zhao, Weigang; Zhang, Ruixia; Min, Qin; Peng, Song

2013-01-01

A confocal micro-X-ray fluorescence (MXRF) technology based on polycapillary X-ray optics was proposed for determining surface topography. This confocal topography method involves elemental sensitivity and can be used to classify the objects according to their elemental composition while obtaining their surface topography. To improve the spatial resolution of this confocal topography technology, the center of the confocal micro-volume was overlapped with the output focal spot of the polycapillary X-ray, focusing the lens in the excitation channel. The input focal spot of the X-ray lens parallel to the detection channel was used to determine the surface position of the sample. The corresponding surface adaptive algorithm was designed to obtain the surface topography. The surface topography of a ceramic chip was obtained. This confocal MXRF surface topography method could find application in the materials sciences

Plasma production and heating by a laser TEA-CO2

International Nuclear Information System (INIS)

Goes, L.C.S.; Sudano, J.P.; Rodrigues, N.A.S.

1987-01-01

Preliminary experiments of plasma production and heating by laser irradiation of gases and solid targets have been performed with a laser TEA-CO 2 (1 MW, 80 ns, monomode), developed and built at the IEAv/Laser Laboratory. The laser beam was focused in the interior of a vacuum chamber (100 1) with a base pressure of 10 1 torr, and recolimated by a system of confocal lenses. The breakdown theresholds for nitrogen gas was investigated by varying the laser power, the neutral gas density and the focal lenght of the lenses. Plasma breakdown observed in the range of pressures between 100-720 torr was in good agreement with calculations of cascade ionization theory and classical absorption by inverse-Bremsstrahlung. The laser absorption was inferred by measuring the power transmitted in the presence and absence of plasma. The light emitted by the plasma was detected by a fast photo-diode, indicating that the plasma expansion phase lasted for several microseconds. These investigations have been applied in the development of plasma shutters for laser pulse compression. (author) [pt

The Use of Intravital Two-Photon and Thick Section Confocal Imaging to Analyze B Lymphocyte Trafficking in Lymph Nodes and Spleen.

Science.gov (United States)

Park, Chung; Hwang, Il-Young; Kehrl, John H

2018-01-01

Intravital two-photon laser scanning microscopy (TP-LSM) has allowed the direct observation of immune cells in intact organs of living animals. In the B cell biology field TP-LSM has detailed the movement of B cells in high endothelial venules and during their transmigration into lymph organs; described the movement and positioning of B cells within lymphoid organs; outlined the mechanisms by which antigen is delivered to B cells; observed B cell interacting with T cells, other cell types, and even with pathogens; and delineated the egress of B cells from the lymph node (LN) parenchyma into the efferent lymphatics. As the quality of TP-LSM improves and as new fluorescent probes become available additional insights into B cell behavior and function await new investigations. Yet intravital TP-LSM has some disadvantages including a lower resolution than standard confocal microscopy, a narrow imaging window, and a shallow depth of imaging. We have found that supplementing intravital TP-LSM with conventional confocal microscopy using thick LN sections helps to overcome some of these shortcomings. Here, we describe procedures for visualizing the behavior and trafficking of fluorescently labeled, adoptively transferred antigen-activated B cells within the inguinal LN of live mice using two-photon microscopy. Also, we introduce procedures for fixed thick section imaging using standard confocal microscopy, which allows imaging of fluorescently labeled cells deep in the LN cortex and in the spleen with high resolution.

Depth-profiling by confocal Raman microscopy (CRM): data correction by numerical techniques.

Science.gov (United States)

Tomba, J Pablo; Eliçabe, Guillermo E; Miguel, María de la Paz; Perez, Claudio J

2011-03-01

The data obtained in confocal Raman microscopy (CRM) depth profiling experiments with dry optics are subjected to significant distortions, including an artificial compression of the depth scale, due to the combined influence of diffraction, refraction, and instrumental effects that operate on the measurement. This work explores the use of (1) regularized deconvolution and (2) the application of simple rescaling of the depth scale as methodologies to obtain an improved, more precise, confocal response. The deconvolution scheme is based on a simple predictive model for depth resolution and the use of regularization techniques to minimize the dramatic oscillations in the recovered response typical of problem inversion. That scheme is first evaluated using computer simulations on situations that reproduce smooth and sharp sample transitions between two materials and finally it is applied to correct genuine experimental data, obtained in this case from a sharp transition (planar interface) between two polymeric materials. It is shown that the methodology recovers very well most of the lost profile features in all the analyzed situations. The use of simple rescaling appears to be only useful for correcting smooth transitions, particularly those extended over distances larger than those spanned by the operative depth resolution, which limits the strategy to the study of profiles near the sample surface. However, through computer simulations, it is shown that the use of water immersion objectives may help to reduce optical distortions and to expand the application window of this simple methodology, which could be useful, for instance, to safely monitor Fickean sorption/desorption of penetrants in polymer films/coatings in a nearly noninvasive way.

Speckle-illuminated fluorescence confocal microscopy, using a digital micro-mirror device

International Nuclear Information System (INIS)

Jiang, Shi-Hong; Walker, John G

2009-01-01

An implementation of a speckle-illuminated fluorescence confocal microscope using a digital micro-mirror device (DMD) is described. The DMD not only projects a sequence of imaged binary speckle patterns onto the specimen at a very high frame rate but also operates as a spatial light modulator to perform real-time optical data processing. Frame averaging is accomplished by CCD charge accumulation during a single exposure. The recorded time-averaged image is a confocal image plus an unwanted non-confocal image which can be removed by recording a separate image. Experimental results with image acquisition within a fraction of a second are shown. Images of a thin biological sample are also shown to demonstrate practical application of the technique

Fungal keratitis - improving diagnostics by confocal microscopy

DEFF Research Database (Denmark)

Nielsen, Esben; Heegaard, S; Prause, J U

2013-01-01

Purpose: Introducing a simple image grading system to support the interpretation of in vivo confocal microscopy (IVCM) images in filamentous fungal keratitis. Setting: Clinical and confocal studies took place at the Department of Ophthalmology, Aarhus University Hospital, Denmark. Histopathological...... analysis was performed at the Eye Pathology Institute, Department of Neuroscience and Pharmacology, University of Copenhagen, Denmark. Methods: A recent series of consecutive patients with filamentous fungal keratitis is presented to demonstrate the results from in-house IVCM. Based upon our experience...... with IVCM and previously published images, we composed a grading system for interpreting IVCM images of filamentous fungal keratitis. Results: A recent case series of filamentous fungal keratitis from 2011 to 2012 was examined. There were 3 male and 3 female patients. Mean age was 44.5 years (range 12...

Morphological change study on root surfaces treated with curettes, sonic instruments or Er:YAG laser; Estudo in vitro da alteracao morfologica em superficie radicular tratada com curetas, aparelho ultrasonico ou com laser de Er:YAG

Energy Technology Data Exchange (ETDEWEB)

Guimaraes Filho, Arlindo Lopes

2004-07-01

Periodontal disease is caused by dental plaque and dental calculus on roots surfaces, specially on cervical areas. As dental plaque is the main cause and dental calculus a secondary one, it is practically impossible to separate one factor to the other one. In order to get periodontal tissue health it is necessary to eliminate dental plaque and calculus from root surfaces. In this sense, Er:YAG laser comes in as an excellent way to control periodontal disease, not only, by removing calculus and dental plaque but also for its bacteria reduction. The aim of this study is to compare, by S.E.M., root surfaces changing when they are treated with curettes and ultrasonic scaling or Er:YAG laser irradiation with two different energy levels of 60 mJ/pulse and 100 mJ/pulse and repetition tax of 10 Hz (in the display). It is also objective of this study to check a possible thermic damage to pulp tissue when the roots surfaces are irradiated with Er:YAG laser. We used for this study, five human dental roots, each one of them were cut into four samples, giving us a total of twenty samples, which were divided in five groups of four samples each one. The control group, we did not indicated any kind of treatment. The first group, the roots samples were scaled and planned with Gracey curettes 5/6 and 7/8. The second group, the roots samples were treated with ultrasonic instruments. The third group was irradiated with Er:YAG laser using 60 mJ/pulse , 10 Hz and energy density of 4 J/cm{sup 2} (approximated). The fourth group was irradiated with Er:YAG laser using 100 mJ/pulse, 10 Hz and energy density of 7 J/cm{sup 2} (approximated). The results analysis showed that roots scaling either with Gracey curettes or with ultrasonic instruments created smear layer covering roots surfaces; roots surfaces irradiated with Er:YAG laser showed few roughness in the third group; roots surfaces irradiated with Er:YAG laser showed no smear layer and the Er:YAG laser irradiation did not bring any

Análisis fractográfico de fibras de circona y de zafiro mediante microscopía óptica confocal

Directory of Open Access Journals (Sweden)

López-Cepero, J. M.

2005-08-01

Full Text Available Fractography is a very useful tool to learn about the mechanisms controlling the fracture process. In this work, the fractographical uses of laser scanning confocal microscopy (LSCM are shown. LSCM is a widely used technique in Biology and similar disciplines, but its use in Materials Science is not yet as explored. However, it is an ideal technique for fractographical studies, since by using it the three-dimensional profile of the fracture surface can be obtained. From this profile, it is possible to extract information, like the roughness of the fracture surface, which would be very difficult to obtain from other studies. In this paper, LSCM is applied to the study of the fracture surface in ceramic fibers submitted to tensile stress, making the interest of the technique evident due to features such as easy sample preparation, gathering of real 3D information, and good SEM-LSCM synergy.

La fractografía en materiales resulta de gran utilidad para la caracterización de los mecanismos que dominan la rotura. En el presente artículo se investigan las aplicaciones fractográficas de la microscopía óptica confocal o LSCM (Laser Scanning Confocal Microscopy. Dicha técnica es de amplio uso en Biología y disciplinas afines, pero su empleo en Ciencia de Materiales está poco explorado. Sin embargo, resulta ideal para el estudio fractográfico, ya que permite obtener reconstrucciones del perfil tridimensional de la superficie de fractura. De este perfil tridimensional puede extraerse información —la rugosidad de la superficie, por ejemplo— muy difícil de obtener a partir de otro tipo de estudios. En este trabajo se aplica LSCM a la caracterización de superficies de fractura en fibras cerámicas sometidas a tracción y se pone de manifiesto el interés de la técnica, debido a características tales como la preparación sencilla de muestras, la obtención de información tridimensional real y la buena coordinación SEM-LSCM.

Gastric Tissue Damage Analysis Generated by Ischemia: Bioimpedance, Confocal Endomicroscopy, and Light Microscopy

Directory of Open Access Journals (Sweden)

Nohra E. Beltran

2013-01-01

Full Text Available The gastric mucosa ischemic tissular damage plays an important role in critical care patients’ outcome, because it is the first damaged tissue by compensatory mechanism during shock. The aim of the study is to relate bioimpedance changes with tissular damage level generated by ischemia by means of confocal endomicroscopy and light microscopy. Bioimpedance of the gastric mucosa and confocal images were obtained from Wistar male rats during basal and ischemia conditions. They were anesthetized, and stain was applied (fluorescein and/or acriflavine. The impedance spectroscopy catheter was inserted and then confocal endomicroscopy probe. After basal measurements and biopsy, hepatic and gastric arteries clamping induced ischemia. Finally, pyloric antrum tissue was preserved in buffered formaldehyde (10% for histology processing using light microscopy. Confocal images were equalized, binarized, and boundary defined, and infiltrations were quantified. Impedance and infiltrations increased with ischemia showing significant changes between basal and ischemia conditions (. Light microscopy analysis allows detection of general alterations in cellular and tissular integrity, confirming gastric reactance and confocal images quantification increments obtained during ischemia.

Microscopia confocal in vivo na cistinose: relato de caso

Directory of Open Access Journals (Sweden)

Victor Gustavo

2004-01-01

Full Text Available A cistinose é doença autossômica recessiva rara caracterizada pelo acúmulo do aminoácido cistina livre dentro dos lisossomos e geralmente é fatal na primeira década de vida na ausência de transplante renal. O presente estudo tem por objetivo relatar os achados da microscopia confocal in vivo em paciente adulto com cistinose infantil. O exame de microscopia confocal in vivo revelou que há diferenças quanto à intensidade de acometimento, tamanho e forma dos depósitos nas diversas camadas corneanas.

Fluorescence confocal polarizing microscopy: Three-dimensional ...

Indian Academy of Sciences (India)

journal of. August 2003 physics pp. 373–384. Fluorescence confocal polarizing ... and focal conic domains in flat samples of lamellar LCs are practically indistinguishable. ... or less) LC layer confined between two transparent plates. ... in studies of electro-optic effects such as the Frederiks effect, defects, surface anchoring,.

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

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.

In-class didactic versus self-directed teaching of the probe-based confocal laser endomicroscopy (pCLE) criteria for Barrett's esophagus.

Science.gov (United States)

Rzouq, Fadi; Vennalaganti, Prashanth; Pakseresht, Kavous; Kanakadandi, Vijay; Parasa, Sravanthi; Mathur, Sharad C; Alsop, Benjamin R; Hornung, Benjamin; Gupta, Neil; Sharma, Prateek

2016-02-01

Optimal teaching methods for disease recognition using probe-based confocal laser endomicroscopy (pCLE) have not been developed. Our aim was to compare in-class didactic teaching vs. self-directed teaching of Barrett's neoplasia diagnosis using pCLE. This randomized controlled trial was conducted at a tertiary academic center. Study participants with no prior pCLE experience were randomized to in-class didactic (group 1) or self-directed teaching groups (group 2). For group 1, an expert conducted a classroom teaching session using standardized educational material. Participants in group 2 were provided with the same material on an audio PowerPoint. After initial training, all participants graded an initial set of 20 pCLE videos and reviewed correct responses with the expert (group 1) or on audio PowerPoint (group 2). Finally, all participants completed interpretations of a further 40 videos. Eighteen trainees (8 medical students, 10 gastroenterology trainees) participated in the study. Overall diagnostic accuracy for neoplasia prediction by pCLE was 77 % (95 % confidence interval [CI] 74.0 % - 79.2 %); of predictions made with high confidence (53 %), the accuracy was 85 % (95 %CI 81.8 % - 87.8 %). The overall accuracy and interobserver agreement was significantly higher in group 1 than in group 2 for all predictions (80.4 % vs. 73 %; P = 0.005) and for high confidence predictions (90 % vs. 80 %; P didactic teaching enables significantly better recognition of the pCLE features of Barrett's esophagus than self-directed teaching. The in-class didactic group had a shorter learning curve and were able to achieve 90 % accuracy for their high confidence predictions. © Georg Thieme Verlag KG Stuttgart · New York.

Characterisation of an inlet pre-injector laser-induced fluorescence instrument for the measurement of atmospheric hydroxyl radicals

Science.gov (United States)

Novelli, A.; Hens, K.; Tatum Ernest, C.; Kubistin, D.; Regelin, E.; Elste, T.; Plass-Dülmer, C.; Martinez, M.; Lelieveld, J.; Harder, H.

2014-10-01

Atmospheric measurements of hydroxyl radicals (OH) are challenging due to a high reactivity and consequently low concentration. The importance of OH as an atmospheric oxidant has motivated a sustained effort leading to the development of a number of highly sensitive analytical techniques. Recent work has indicated that the laser-induced fluorescence of the OH molecules method based on the fluorescence assay by gas expansion technique (LIF-FAGE) for the measurement of atmospheric OH in some environments may be influenced by artificial OH generated within the instrument, and a chemical method to remove this interference was implemented in a LIF-FAGE system by Mao et al. (2012). While it is not clear whether other LIF-FAGE instruments suffer from the same interference, we have applied this method to our LIF-FAGE HORUS (Hydroxyl Radical Measurement Unit based on fluorescence Spectroscopy) system, and developed and deployed an inlet pre-injector (IPI) to determine the chemical zero level in the instrument via scavenging the ambient OH radical. We describe and characterise this technique in addition to its application at field sites in forested locations in Finland, Spain and Germany. Ambient measurements show that OH generated within the HORUS instrument is a non-negligible fraction of the total OH signal, which can comprise 30 to 80% during daytime and 60 to 100% during the night. The contribution of the background OH varied greatly between measurement sites and was likely related to the type and concentration of volatile organic compounds (VOCs) present at each particular location. Two inter-comparisons in contrasting environments between the HORUS instrument and two different chemical ionisation mass spectrometers (CIMS) are described to demonstrate the efficacy of IPI and the necessity of the chemical zeroing method for our LIF-FAGE instrument in such environments.

Surface wettability of silicon substrates enhanced by laser ablation

Energy Technology Data Exchange (ETDEWEB)

Tseng, Shih-Feng [National Applied Research Laboratories, Instrument Technology Research Center, Hsinchu (China); National Chiao Tung University, Department of Mechanical Engineering, Hsinchu (China); Hsiao, Wen-Tse; Huang, Kuo-Cheng; Hsiao, Sheng-Yi [National Applied Research Laboratories, Instrument Technology Research Center, Hsinchu (China); Chen, Ming-Fei [National Changhua University of Education, Department of Mechatronics Engineering, Changhua (China); Lin, Yung-Sheng [Hungkuang University, Department of Applied Cosmetology and Graduate Institute of Cosmetic Science, Taichung (China); Chou, Chang-Pin [National Chiao Tung University, Department of Mechanical Engineering, Hsinchu (China)

2010-11-15

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

Classifying distinct basal cell carcinoma subtype by means of dermatoscopy and reflectance confocal microscopy.

Science.gov (United States)

Longo, Caterina; Lallas, Aimilios; Kyrgidis, Athanassios; Rabinovitz, Harold; Moscarella, Elvira; Ciardo, Silvana; Zalaudek, Iris; Oliviero, Margaret; Losi, Amanda; Gonzalez, Salvador; Guitera, Pascale; Piana, Simonetta; Argenziano, Giuseppe; Pellacani, Giovanni

2014-10-01

The current guidelines for the management of basal cell carcinoma (BCC) suggest a different therapeutic approach according to histopathologic subtype. Although dermatoscopic and confocal criteria of BCC have been investigated, no specific studies were performed to evaluate the distinct reflectance confocal microscopy (RCM) aspects of BCC subtypes. To define the specific dermatoscopic and confocal criteria for delineating different BCC subtypes. Dermatoscopic and confocal images of histopathologically confirmed BCCs were retrospectively evaluated for the presence of predefined criteria. Frequencies of dermatoscopic and confocal parameters are provided. Univariate and adjusted odds ratios were calculated. Discriminant analyses were performed to define the independent confocal criteria for distinct BCC subtypes. Eighty-eight BCCs were included. Dermatoscopically, superficial BCCs (n=44) were primarily typified by the presence of fine telangiectasia, multiple erosions, leaf-like structures, and revealed cords connected to the epidermis and epidermal streaming upon RCM. Nodular BCCs (n=22) featured the classic dermatoscopic features and well outlined large basaloid islands upon RCM. Infiltrative BCCs (n=22) featured structureless, shiny red areas, fine telangiectasia, and arborizing vessels on dermatoscopy and dark silhouettes upon RCM. The retrospective design. Dermatoscopy and confocal microscopy can reliably classify different BCC subtypes. Copyright © 2014 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.

Spatially selective Er/Yb-doped CaF2 crystal formation by CO2 laser exposure

International Nuclear Information System (INIS)

Kim, Dong-Seon; Lee, Jin-Ho; Lim, Ki-Soo

2014-01-01

Highlights: • Oxyfluoride glass–ceramics containing CaF 2 nanocrystals doped with Er 3+ and Yb 3+ ions were formed on the glass surface by CO 2 laser and a heat gun exposure. • Most of Er and Yb ions were distributed inside CaF 2 nanocrystals and fluorine loss was observed in the EDS element maps. • IR-to-VIS upconversion emission efficiency of laser annealed glass ceramics was much increased and compared with that of the furnace-annealed glass ceramics. • Distributed volume of the glass ceramics were estimated by a confocal fluorescence microscope imaging. - Abstract: We report the glass–ceramic precipitation on the oxyfluoride glass surface by spatially selective annealing with a CO 2 laser and a heat gun exposure. X-ray diffraction analysis showed the formation of major CaF 2 and miner Ca 2 SiO 4 nanoparticles. We observed ∼100 nm nanoparticle aggregation by tunneling electron microscopy and element distribution in glass and crystal phases. Spatial distribution of glass ceramics near the glass surface was probed by confocal fluorescence microscope by using much enhanced emission from the Er ions in the laser-treated area. Strong emissions at 365 nm excitation and visible up-conversion emissions at 980 nm excitation also indicated well incorporation of Er and Yb ions into a crystalline environment

Spatially selective Er/Yb-doped CaF2 crystal formation by CO2 laser exposure

International Nuclear Information System (INIS)

Kim, Dong-Seon; Lee, Jin-Ho; Lim, Ki-Soo

2015-01-01

Highlights: • Oxyfluoride glass–ceramics containing CaF 2 nanocrystals doped with Er 3+ and Yb 3+ ions were formed on the glass surface by CO 2 laser and a heat gun exposure. • Most of Er and Yb ions were distributed inside CaF 2 nanocrystals and fluorine loss was observed in the EDS element maps. • IR-to-VIS upconversion emission efficiency of laser annealed glass ceramics was much increased and compared with that of the furnace-annealed glass ceramics. • Distributed volume of the glass ceramics were estimated by a confocal fluorescence microscope imaging. - Abstract: We report the glass–ceramic precipitation on the oxyfluoride glass surface by spatially selective annealing with a CO 2 laser and a heat gun exposure. X-ray diffraction analysis showed the formation of major CaF 2 and miner Ca 2 SiO 4 nanoparticles. We observed ∼100 nm nanoparticle aggregation by tunneling electron microscopy and element distribution in glass and crystal phases. Spatial distribution of glass ceramics near the glass surface was probed by confocal fluorescence microscope by using much enhanced emission from the Er ions in the laser-treated area. Strong emissions at 365 nm excitation and visible up-conversion emissions at 980 nm excitation also indicated well incorporation of Er and Yb ions into a crystalline environment

Laser polarimetry

International Nuclear Information System (INIS)

Goldstein, D.H.

1989-01-01

Polarimetry, or transmission ellipsometry, is an important experimental technique for the determination of polarization properties of bulk materials. In this technique, source radiation of known polarization is passed through bulk samples to determine, for example, natural or induced birefringence and dichroism. The laser is a particularly appropriate source for this technique because of its monochromaticity, collimation, and radiant intensity. Lasers of many different wavelengths in different spectral regions are now available. Laser polarimetry can be done in any of these wavelength regions where polarizing elements are available. In this paper, polarimetry is reviewed with respect to applications, sources used, and polarization state generator and analyzer configurations. Scattering ellipsometry is also discussed insofar as the forward scattering measurement is related to polarimetry. The authors then describe an infrared laser polarimeter which we have designed and constructed. This instrument can operate over large wavelength regions with only a change in source. Polarization elements of the polarimeter are in a dual rotating retarder configuration. Computer controlled rotary stages and computer monitored detectors automate the data collection. The Mueller formulation is used to process the polarization information. Issues and recent progress with this instrument are discussed

Assessing delivery and quantifying efficacy of small interfering ribonucleic acid therapeutics in the skin using a dual-axis confocal microscope

Science.gov (United States)

Ra, Hyejun; Gonzalez-Gonzalez, Emilio; Smith, Bryan R.; Gambhir, Sanjiv S.; Kino, Gordon S.; Solgaard, Olav; Kaspar, Roger L.; Contag, Christopher H.

2010-05-01

Transgenic reporter mice and advances in imaging instrumentation are enabling real-time visualization of cellular mechanisms in living subjects and accelerating the development of novel therapies. Innovative confocal microscope designs are improving their utility for microscopic imaging of fluorescent reporters in living animals. We develop dual-axis confocal (DAC) microscopes for such in vivo studies and create mouse models where fluorescent proteins are expressed in the skin for the purpose of advancing skin therapeutics and transdermal delivery tools. Three-dimensional image volumes, through the different skin compartments of the epidermis and dermis, can be acquired in several seconds with the DAC microscope in living mice, and are comparable to histologic analyses of reporter protein expression patterns in skin sections. Intravital imaging with the DAC microscope further enables visualization of green fluorescent protein (GFP) reporter gene expression in the skin over time, and quantification of transdermal delivery of small interfering RNA (siRNA) and therapeutic efficacy. Visualization of transdermal delivery of nucleic acids will play an important role in the development of innovative strategies for treating skin pathologies.

Proceedings of national laser symposium (NLS-2000)

International Nuclear Information System (INIS)

Mallik, Amitav; Srivastava, K.N.; Pal, Suranjan

2000-01-01

This proceedings comprise of a series of invited talks on selected topics in lasers and wide range of contributed papers. The main topics are laser physics and research, laser devices and technology, laser materials and spectroscopy, quantum optics, non-linear optics ultra-fast phenomenon, laser produced plasma, high power lasers, laser instrumentation, medical applications and industrial applications of lasers and fiber optics. The papers relevant to INIS Database are indexed separately

Lasers in periodontics.

Science.gov (United States)

Elavarasu, Sugumari; Naveen, Devisree; Thangavelu, Arthiie

2012-08-01

Laser is one of the most captivating technologies in dental practice since Theodore Maiman in 1960 invented the ruby laser. Lasers in dentistry have revolutionized several areas of treatment in the last three and a half decades of the 20(th) century. Introduced as an alternative to mechanical cutting device, laser has now become an instrument of choice in many dental applications. Evidence suggests its use in initial periodontal therapy, surgery, and more recently, its utility in salvaging implant opens up a wide range of applications. More research with better designs are a necessity before lasers can become a part of dental armamentarium. This paper gives an insight to laser in periodontics.

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

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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.

Latest developments and opportunities for 3D analysis of biological samples by confocal μ-XRF

International Nuclear Information System (INIS)

Perez, Roberto D.; Sanchez, Hector J.; Perez, Carlos A.; Rubio, Marcelo

2010-01-01

X-ray fluorescence analysis performed with a primary radiation focused in the micrometer range is known as micro-X-ray fluorescence (μ-XRF). It is characterized by a penetration depth higher than other micro-analytical methods, reaching hundreds of micrometers in biological samples. This characteristic of the X-ray beam can be employed in 3D analysis. An innovative method to perform 3D analysis by μ-XRF is the so-called confocal setup. The confocal setup consists of X-ray lenses in the excitation as well as in the detection channel. In this configuration, a micro-volume defined by the overlap of the foci of both X-ray lenses is analyzed. Scanning this micro-volume through the sample can be used to perform a study in three dimensions. At present, X-ray lenses used in confocal μ-XRF experiments are mainly glass capillaries and polycapillaries. Glass capillaries are used in the excitation channel with sources of high photon flux like synchrotron radiation. Half polycapillaries or conical polycapillary concentrators are used almost exclusively in the detection channel. Spatial resolution of the confocal μ-XRF depends on the dimensions of the foci of both X-ray lenses. The overlap of these foci forms an ellipsoid which is the probing volume of the confocal setup. The axis length of the probing volume reported in confocal μ-XRF experiments are of order of few tens of micrometer. In our confocal setup, we used a commercial glass monocapillary in the excitation channel and a monolithic half polycapillary in the detection channel. The polycapillary was home-made by means of drawing of multibundles of glass capillaries in a heating furnace. The experiment was carried out at the beamline D09B-XRF of the Synchrotron Light National Laboratory (Laboratorio Nacional de Luz Sincrotron, LNLS) using white beam. A model for the theoretical description of X-ray fluorescence intensity registered by confocal μ-XRF was introduced by Malzer and Kanngieβer [2005. A model for the

Measurement and quantification of fluorescent changes in ocular tissue using a novel confocal instrument

Science.gov (United States)

Buttenschoen, Kim K.; Girkin, John M.; Daly, Daniel J.

2014-05-01

Our sight is a major contributor to our quality of life. The treatment of diseases like macular degeneration and glaucoma, however, presents a challenge as the delivery of medication to ocular tissue is not well understood. The instrument described here will help quantify targeted delivery by non-invasively and simultaneously measuring light reflected from and fluorescence excited in the eye, used as position marker and to track compounds respectively. The measurement concept has been proven by monitoring the diffusion of fluorescein and a pharmaceutical compound for treating open angle glaucoma in vitro in a cuvette and in ex vivo porcine eyes. To obtain a baseline of natural fluorescence we measured the change in corneal and crystalline lens autofluorescence in volunteers over a week. We furthermore present data on 3D ocular autofluorescence. Our results demonstrate the capability to measure the location and concentration of the compound of interest with high axial and temporal resolution of 178 μm and 0.6 s respectively. The current detection limit is 2 nM for fluorescein, and compounds with a quantum yield as low as 0.01 were measured to concentrations below 1 μM. The instrument has many applications in assessing the diffusion of fluorescent compounds through the eye and skin in vitro and in vivo, measuring autofluorescence of ocular tissues and reducing the number of animals needed for research. The instrument has the capability of being used both in the clinical and home care environment opening up the possibility of measuring controlled drug release in a patient friendly manner.

Fluorescence Confocal Microscopy for Ex Vivo Diagnosis of Conjunctival Tumors: A Pilot Study.

Science.gov (United States)

Iovieno, Alfonso; Longo, Caterina; De Luca, Mariacarla; Piana, Simonetta; Fontana, Luigi; Ragazzi, Moira

2016-08-01

To evaluate the potential use of fluorescence confocal microscopy (FCM) for ex vivo diagnosis and excision margin assessment of conjunctival neoplasms. Validity study. setting: Single institution. Consecutive patients with clinically suspicious conjunctival lesions. Conjunctival lesions were excised in toto using a standard "no-touch technique" by a single surgeon (A.I.). Collected specimens were examined with a commercially available laser scanning fluorescence confocal microscope after immersion in a 0.6 mM solution of acridine orange dye for 10-20 seconds. Specimens were subsequently processed with standard histologic analysis. FCM diagnosis of the nature and extension of conjunctival lesions. Sixteen consecutive patients were included in the study (11 male, 5 female; mean age 58.1 ± 26.1 years, range 10-90 years). The median time needed to process and analyze a sample with FCM was 15 minutes. Eleven of 16 lesions were identified by FCM as squamous (2 benign papillomas, 2 grade 2 conjunctival intraepithelial neoplasias, 7 in situ squamous carcinomas) and 5 as nonsquamous (1 pingueculum, 1 dermolipoma, 2 melanocytic nevi, 1 melanoma). In all cases FCM was able to detect horizontal and vertical extension of the lesion. All FCM findings were confirmed by corresponding subsequent histologic examination. FCM provides a fast ex vivo preliminary diagnosis of suspicious conjunctival lesions with good histologic details and margin assessment, and may represent a novel tool for intraoperative and postsurgical management of conjunctival tumors. This is the first study to investigate ex vivo FCM application in ophthalmology. Copyright © 2016 Elsevier Inc. All rights reserved.

Methods of Hematoxylin and Erosin Image Information Acquisition and Optimization in Confocal Microscopy.

Science.gov (United States)

Yoon, Woong Bae; Kim, Hyunjin; Kim, Kwang Gi; Choi, Yongdoo; Chang, Hee Jin; Sohn, Dae Kyung

2016-07-01

We produced hematoxylin and eosin (H&E) staining-like color images by using confocal laser scanning microscopy (CLSM), which can obtain the same or more information in comparison to conventional tissue staining. We improved images by using several image converting techniques, including morphological methods, color space conversion methods, and segmentation methods. An image obtained after image processing showed coloring very similar to that in images produced by H&E staining, and it is advantageous to conduct analysis through fluorescent dye imaging and microscopy rather than analysis based on single microscopic imaging. The colors used in CLSM are different from those seen in H&E staining, which is the method most widely used for pathologic diagnosis and is familiar to pathologists. Computer technology can facilitate the conversion of images by CLSM to be very similar to H&E staining images. We believe that the technique used in this study has great potential for application in clinical tissue analysis.

Ablation mass features in multi-pulses femtosecond laser ablate molybdenum target

Science.gov (United States)

Zhao, Dongye; Gierse, Niels; Wegner, Julian; Pretzler, Georg; Oelmann, Jannis; Brezinsek, Sebastijan; Liang, Yunfeng; Neubauer, Olaf; Rasinski, Marcin; Linsmeier, Christian; Ding, Hongbin

2018-03-01

In this study, the ablation mass features related to reflectivity of bulk Molybdenum (Mo) were investigated by a Ti: Sa 6 fs laser pulse at central wavelength 790 nm. The ablated mass removal was determined using Confocal Microscopy (CM) technique. The surface reflectivity was calibrated and measured by a Lambda 950 spectrophotometer as well as a CCD camera during laser ablation. The ablation mass loss per pulse increase with the increasing of laser shots, meanwhile the surface reflectivity decrease. The multi-pulses (100 shots) ablation threshold of Mo was determined to be 0.15 J/cm2. The incubation coefficient was estimated as 0.835. The reflectivity change of the Mo target surface following multi-pulses laser ablation were studied as a function of laser ablation shots at various laser fluences from 1.07 J/cm2 to 36.23 J/cm2. The results of measured reflectivity indicate that surface reflectivity of Mo target has a significant decline in the first 3-laser pulses at the various fluences. These results are important for developing a quantitative analysis model for laser induced ablation and laser induced breakdown spectroscopy for the first wall diagnosis of EAST tokamak.

Color Laser Microscope

Science.gov (United States)

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

1987-04-01

A color laser microscope utilizing a new color laser imaging system has been developed for the visual inspection of semiconductors. The light source, produced by three lasers (Red; He-Ne, Green; Ar, Blue; He-Cd), is deflected horizontally by an AOD (Acoustic Optical Deflector) and vertically by a vibration mirror. The laser beam is focused in a small spot which is scanned over the sample at high speed. The light reflected back from the sample is reformed to contain linear information by returning to the original vibration mirror. The linear light is guided to the CCD image sensor where it is converted into a video signal. Individual CCD image sensors are used for each of the three R, G, or B color image signals. The confocal optical system with its laser light source yields a color TV monitor image with no flaring and a much sharper resolution than that of the conventional optical microscope. The AOD makes possible a high speed laser scan and a NTSC or PAL TV video signal is produced in real time without any video memory. Since the light source is composed of R, G, and B laser beams, color separation superior to that of white light illumination is achieved. Because of the photometric linearity of the image detector, the R, G, and B outputs of the system are most suitably used for hue analysis. The CCD linear image sensors in the optical system produce no geometrical distortion, and good color registration is available principally. The output signal can be used for high accuracy line width measuring. The many features of the color laser microscope make it ideally suited for the visual inspection of semiconductor processing. A number of these systems have already been installed in such a capacity. The Color Laser Microscope can also be a very useful tool for the fields of material engineering and biotechnology.

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

Quantification of confocal fluorescence microscopy for the detection of cervical intraepithelial neoplasia.

Science.gov (United States)

Sheikhzadeh, Fahime; Ward, Rabab K; Carraro, Anita; Chen, Zhao Yang; van Niekerk, Dirk; Miller, Dianne; Ehlen, Tom; MacAulay, Calum E; Follen, Michele; Lane, Pierre M; Guillaud, Martial

2015-10-24

Cervical cancer remains a major health problem, especially in developing countries. Colposcopic examination is used to detect high-grade lesions in patients with a history of abnormal pap smears. New technologies are needed to improve the sensitivity and specificity of this technique. We propose to test the potential of fluorescence confocal microscopy to identify high-grade lesions. We examined the quantification of ex vivo confocal fluorescence microscopy to differentiate among normal cervical tissue, low-grade Cervical Intraepithelial Neoplasia (CIN), and high-grade CIN. We sought to (1) quantify nuclear morphology and tissue architecture features by analyzing images of cervical biopsies; and (2) determine the accuracy of high-grade CIN detection via confocal microscopy relative to the accuracy of detection by colposcopic impression. Forty-six biopsies obtained from colposcopically normal and abnormal cervical sites were evaluated. Confocal images were acquired at different depths from the epithelial surface and histological images were analyzed using in-house software. The features calculated from the confocal images compared well with those features obtained from the histological images and histopathological reviews of the specimens (obtained by a gynecologic pathologist). The correlations between two of these features (the nuclear-cytoplasmic ratio and the average of three nearest Delaunay-neighbors distance) and the grade of dysplasia were higher than that of colposcopic impression. The sensitivity of detecting high-grade dysplasia by analysing images collected at the surface of the epithelium, and at 15 and 30 μm below the epithelial surface were respectively 100, 100, and 92 %. Quantitative analysis of confocal fluorescence images showed its capacity for discriminating high-grade CIN lesions vs. low-grade CIN lesions and normal tissues, at different depth of imaging. This approach could be used to help clinicians identify high-grade CIN in clinical

MOONLIGHT: A NEW LUNAR LASER RANGING RETROREFLECTOR INSTRUMENT

Directory of Open Access Journals (Sweden)

M. Garattini

2013-12-01

Full Text Available Since 1969 Lunar Laser Ranging (LLR to the Apollo Cube Corner Reflector (CCR arrays has supplied several significant tests of gravity: Geodetic Precession, the Strong and Weak Equivalence Principle (SEP, WEP, the Parametrized Post Newtonian (PPN parameter , the time change of the Gravitational constant (G, 1/r2 deviations and new gravitational theories beyond General Relativity (GR, like the unified braneworld theory (G. Dvali et al., 2003. Now a new generation of LLR can do better using evolved laser retroreflectors, developed from tight collaboration between my institution, INFN–LNF (Istituto Nazionale di Fisica Nucleare – Laboratori Nazionali di Frascati, and Douglas Currie (University of Maryland, USA, one of the fathers of LLR. The new lunar CCR is developing and characterizing at the “Satellite/Lunar laser ranging Characterization Facility” (SCF, in Frascati, performing our new industry standard space test procedure, the “SCF-Test”; this work contains the experimental results of the SCF-Test applied to the new lunar CCR, and all the new payload developments, including the future SCF tests. The International Lunar Network (ILN research project considers our new retroreflector as one of the possible “Core Instruments”

3D reconstruction and characterization of laser induced craters by in situ optical microscopy

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