WorldWideScience

Sample records for spinning-disk confocal microscope

  1. Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

    Science.gov (United States)

    Hayashi, Shinichi; Okada, Yasushi

    2015-05-01

    Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging. © 2015 Hayashi and Okada. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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

    Science.gov (United States)

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

    2017-02-01

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

  3. Super-Resolution Imaging Strategies for Cell Biologists Using a Spinning Disk Microscope

    Science.gov (United States)

    Hosny, Neveen A.; Song, Mingying; Connelly, John T.; Ameer-Beg, Simon; Knight, Martin M.; Wheeler, Ann P.

    2013-01-01

    In this study we use a spinning disk confocal microscope (SD) to generate super-resolution images of multiple cellular features from any plane in the cell. We obtain super-resolution images by using stochastic intensity fluctuations of biological probes, combining Photoactivation Light-Microscopy (PALM)/Stochastic Optical Reconstruction Microscopy (STORM) methodologies. We compared different image analysis algorithms for processing super-resolution data to identify the most suitable for analysis of particular cell structures. SOFI was chosen for X and Y and was able to achieve a resolution of ca. 80 nm; however higher resolution was possible >30 nm, dependant on the super-resolution image analysis algorithm used. Our method uses low laser power and fluorescent probes which are available either commercially or through the scientific community, and therefore it is gentle enough for biological imaging. Through comparative studies with structured illumination microscopy (SIM) and widefield epifluorescence imaging we identified that our methodology was advantageous for imaging cellular structures which are not immediately at the cell-substrate interface, which include the nuclear architecture and mitochondria. We have shown that it was possible to obtain two coloured images, which highlights the potential this technique has for high-content screening, imaging of multiple epitopes and live cell imaging. PMID:24130668

  4. 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.

  5. Spinning Disk Confocal System

    National Research Council Canada - National Science Library

    Schoenbach, Karl H

    2006-01-01

    ... of this proposal serving as Pi of the MURI. Experimental studies in which human cells were exposed to pulsed electric fields of up to 300 kV/cm amplitude, with durations as short as 10 ns, have confirmed that these pulses have strong effects...

  6. Spinning Disk Confocal System

    National Research Council Canada - National Science Library

    Schoenbach, Karl H

    2006-01-01

    Radiofrequency (RF) radiation is in the frequency range in which the outer membrane of mammalian cells is no longer a barrier to electric field penetration, allowing access of the RF to subcellular structures...

  7. Spinning Disk Confocal System

    Science.gov (United States)

    2006-06-01

    high temporal resolution. An instrument has been developed for exactly this type of live-cell imaging. This new instrument scans 1000 microbeams across...Imaging System. Instead of scanning a single laser beam across the cell, this new instrument scans 1000 microbeams simultaneously using a spinning...multipoint-excitation, multipoint- emission characteristics of UltraView RS, which confers three main advantages over traditional beam scanning LSCMs for

  8. The use of spinning-disk confocal microscopy for the intravital analysis of platelet dynamics in response to systemic and local inflammation.

    Directory of Open Access Journals (Sweden)

    Craig N Jenne

    Full Text Available Platelets are central players in inflammation and are an important component of the innate immune response. The ability to visualize platelets within the live host is essential to understanding their role in these processes. Past approaches have involved adoptive transfer of labelled platelets, non-specific dyes, or the use of fluorescent antibodies to tag platelets in vivo. Often, these techniques result in either the activation of the platelet, or blockade of specific platelet receptors. In this report, we describe two new methods for intravital visualization of platelet biology, intravenous administration of labelled anti-CD49b, which labels all platelets, and CD41-YFP transgenic mice, in which a percentage of platelets express YFP. Both approaches label endogenous platelets and allow for their visualization using spinning-disk confocal fluorescent microscopy. Following LPS-induced inflammation, we were able to measure a significant increase in both the number and size of platelet aggregates observed within the vasculature of a number of different tissues. Real-time observation of these platelet aggregates reveals them to be large, dynamic structures that are continually expanding and sloughing-off into circulation. Using these techniques, we describe for the first time, platelet recruitment to, and behaviour within numerous tissues of the mouse, both under control conditions and following LPS induced inflammation.

  9. 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

  10. Image inpainting for the differential confocal microscope

    Science.gov (United States)

    Qiu, Lirong; Wang, Lei; Liu, Dali; Hou, Maosheng; Zhao, Weiqian

    2015-02-01

    In the process of zero-crossing trigger measurement of differential confocal microscope, the sample surface features or tilt will cause the edges can't be triggered. Meanwhile, environment vibration can also cause false triggering. In order to restore the invalid information of sample, and realize high-precision surface topography measurement, Total Variation (TV) inpainting model is applied to restore the scanning images. Emulation analysis and experimental verification of this method are investigated. The image inpainting algorithm based on TV model solves the minimization of the energy equation by calculus of variations, and it can effectively restore the non-textured image with noises. Using this algorithm, the simulation confocal laser intensity curve and height curve of standard step sample are restored. After inpainting the intensity curve below the threshold is repaired, the maximum deviation from ideal situation is 0.0042, the corresponding edge contour of height curve is restored, the maximum deviation is 0.1920, which proves the algorithm is effective. Experiment of grating inpainting indicates that the TV algorithm can restore the lost information caused by failed triggering and eliminate the noise caused by false triggering in zero-crossing trigger measurement of differential confocal microscope. The restored image is consistent with the scanning result of OLYMPUS confocal microscope, which can satisfy the request of follow-up measurement analysis.

  11. Acquisition of a Spinning Disk Confocal Microscope to Enhance Research and Teaching Capabilities in the STEM Fields at Howard University

    Science.gov (United States)

    2015-05-13

    Washington, DC. April 16, 2015 Conference: Howard University Research Day 2015 Abstract: Background: The Ceanorhabditis elegans Myt-1 ortholog...the nematode, Ceanorhabditis elegans . One manner in which they characterized the effect of their specific gene knockdown was to determine whether...peer-reviewed journals: (c) Presentations 1) Identifying new players in meiosis using C. elegans depleted of the meiotic inhibitory kinase WEE-1.3

  12. Simultaneous differential spinning disk fluorescence optical sectioning microscopy and nanomechanical mapping atomic force microscopy

    International Nuclear Information System (INIS)

    Miranda, Adelaide; De Beule, Pieter A. A.; Martins, Marco

    2015-01-01

    Combined microscopy techniques offer the life science research community a powerful tool to investigate complex biological systems and their interactions. Here, we present a new combined microscopy platform based on fluorescence optical sectioning microscopy through aperture correlation microscopy with a Differential Spinning Disk (DSD) and nanomechanical mapping with an Atomic Force Microscope (AFM). The illumination scheme of the DSD microscope unit, contrary to standard single or multi-point confocal microscopes, provides a time-independent illumination of the AFM cantilever. This enables a distortion-free simultaneous operation of fluorescence optical sectioning microscopy and atomic force microscopy with standard probes. In this context, we discuss sample heating due to AFM cantilever illumination with fluorescence excitation light. Integration of a DSD fluorescence optical sectioning unit with an AFM platform requires mitigation of mechanical noise transfer of the spinning disk. We identify and present two solutions to almost annul this noise in the AFM measurement process. The new combined microscopy platform is applied to the characterization of a DOPC/DOPS (4:1) lipid structures labelled with a lipophilic cationic indocarbocyanine dye deposited on a mica substrate

  13. A study on improving the resolution of confocal microscope

    International Nuclear Information System (INIS)

    Kim, Young June; Song, Dae Ho; Ryu, Weon Jae

    2001-01-01

    In modern industry, the accuracy and the surface-finish requirements for machined parts have been becoming ever more stringent. Optical method in measurements is used in vibration measurement, crack and defect detection with the advent of opto-mechatronics, and it is expected to play an important role in surface topography. In this study, the principle of confocal microscope is described, and the advanced 3-D surface measuring system that has better measuring speed than the traditional confocal microscope is developed. A study on improving the resolution of the advanced confocal microscope is followed. Finally, Software for data acquisition and analysis of various parameters in surface geometrical features has been developed.

  14. Nanoparticle Imaging with Polarization Interferometric Nonlinear Confocal Microscope

    OpenAIRE

    Fujita, Kohei; Egami, Chikara

    2014-01-01

    Polarization interferometric nonlinear confocal microscope has been developed for single nanoparticle analysis of drug delivery system (DDS). The microscope is a system based on a low cost and low power continuous wave (CW) laser light source. Also, the microscope observed shape anisotropy of the 200 nm diameter nanoparticle. According to nanoparticle imaging and CTF (contrast transfer function) curve observation of the microscope, three-dimensional resolution of the microscope measured up to...

  15. 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...

  16. Corneal Subbasal Nerve Density: A Comparison of Two Confocal Microscopes

    Science.gov (United States)

    Erie, Elizabeth A.; McLaren, Jay W.; Kittleson, Katrina M.; Patel, Sanjay V.; Erie, Jay C.; Bourne, William M.

    2009-01-01

    Purpose To compare subbasal nerve densities estimated from images recorded by the Tandem Scanning and the ConfoScan 4 confocal microscopes. Methods Confocal microscopy was used to estimate subbasal nerve density in 62 corneas of 40 subjects (18 corneas of 18 normal subjects and 44 corneas of 22 patients between 1 and 12 months after LASIK) At each examination, corneas were scanned first by using Tandem Scanning and then by using a ConfoScan 4 confocal microscope. Subbasal nerves from 2 to 4 scans per cornea were traced by using a semi-automated nerve analysis program. Nerve density was expressed as total nerve length divided by the sample area (µm/mm2). Differences in nerve density between instruments were examined by using paired tests. Results In normal corneas, subbasal nerve density was 10,658 ± 5,581 µm/mm2 (mean ± SD) with the ConfoScan 4 and 5,534 ± 1,850 µm/mm2 with the Tandem Scanning microscope (Pmicroscope. These differences must be considered when comparing subbasal nerve densities between studies that use different confocal microscopes. PMID:18997541

  17. Confocal direct imaging Raman microscope: design and applications in biology

    NARCIS (Netherlands)

    Sijtsema, N.M.; Wouters, Siddi D.; de Grauw, Cees J.; de Grauw, C.J.; Otto, Cornelis; Greve, Jan

    1998-01-01

    A confocal direct imaging Raman microscope (CDIRM) based on two synchronized scanning mirrors, a monochromator, and two charge-coupled device (CCD) cameras has been developed. With this system it is possible to make both Raman spectra of a small measurement volume and images of a larger sample area

  18. Modular Scanning Confocal Microscope with Digital Image Processing.

    Science.gov (United States)

    Ye, Xianjun; McCluskey, Matthew D

    2016-01-01

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

  19. 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.

  20. Simplified confocal microscope for counting particles at low concentrations.

    Science.gov (United States)

    Skinner, Joseph P; Swift, Kerry M; Ruan, Qiaoqiao; Perfetto, Sergio; Gratton, Enrico; Tetin, Sergey Y

    2013-07-01

    We describe a compact scanning confocal fluorescence microscope capable of detecting particles concentrations less than 100 particles∕ml in ~15 min. The system mechanically moves a cuvette containing ~3 ml of sample. A relatively large confocal volume is observed within the cuvette using a 1 mm pinhole in front of a detection PMT. Due to the motion of the sample, particles traverse the confocal volume quickly, and analysis by pattern recognition qualifies spikes in the emission intensity data and counts them as events. We show linearity of detection as a function of concentration and also characterize statistical behavior of the instrument. We calculate a detection sensitivity of the system using 3 μm fluorescent microspheres to be 5 particles/ml. Furthermore, to demonstrate biological application, we performed a dilution series to quantify stained E. coli and yeast cells. We counted E. coli cells at a concentration as low as 30 cells∕ml in 10 min/sample.

  1. Confocal Microscope Alignment of Nanocrystals for Coherent Diffraction Imaging

    International Nuclear Information System (INIS)

    Beitra, Loren; Watari, Moyu; Matsuura, Takashi; Shimamoto, Naonobu; Harder, Ross; Robinson, Ian

    2010-01-01

    We have installed and tested an Olympus LEXT confocal microscope at the 34-ID-C beamline of the Advanced Photon Source (APS). The beamline is for Coherent X-ray Diffraction (CXD) experiments in which a nanometre-sized crystal is aligned inside a focussed X-ray beam. The microscope was required for three-dimensional (3D) sample alignment to get around sphere-of-confusion issues when locating Bragg peaks in reciprocal space. In this way, and by use of strategic sample preparations, we have succeeded in measuring six Bragg peaks from a single 200 nm gold crystal and obtained six projections of its internal displacement field. This enables the clear identification of stacking-fault bands within the crystal. The confocal alignment method will allow a full determination of the strain tensor provided three or more Bragg reflections from the same crystal are found.

  2. GFP fluorescence imaging with laser confocal scanning microscope

    Science.gov (United States)

    Yu, Yanhua; Xing, Da; Shi, Qiaojuan; Zhou, Junchu

    1999-09-01

    With gene marking technique, green fluorescent protein (GFP) and nodule bacteria gene has been linked together to form a single fusion gene expression vector. Then the vector is transferred into the nodule bacteria and the astragalus sinicus is invaded by the vector. With laser confocal scanning microscope, some important morphological information in plant nitrogen fixation research about the invading of nodule bacteria and the formation process of root nodule has been obtained through the 3D imaging of GFP reporting fluorescence.

  3. Chromatic confocal microscope using hybrid aspheric diffractive lenses

    Science.gov (United States)

    Rayer, Mathieu; Mansfield, Daniel

    2014-05-01

    A chromatic confocal microscope is a single point non-contact distance measurement sensor. For three decades the vast majority of the chromatic confocal microscope use refractive-based lenses to code the measurement axis chromatically. However, such an approach is limiting the range of applications. In this paper the performance of refractive, diffractive and Hybrid aspheric diffractive are compared. Hybrid aspheric diffractive lenses combine the low geometric aberration of a diffractive lens with the high optical power of an aspheric lens. Hybrid aspheric diffractive lenses can reduce the number of elements in an imaging system significantly or create large hyper- chromatic lenses for sensing applications. In addition, diffractive lenses can improve the resolution and the dynamic range of a chromatic confocal microscope. However, to be suitable for commercial applications, the diffractive optical power must be significant. Therefore, manufacturing such lenses is a challenge. We show in this paper how a theoretical manufacturing model can demonstrate that the hybrid aspheric diffractive configuration with the best performances is achieved by step diffractive surface. The high optical quality of step diffractive surface is then demonstrated experimentally. Publisher's Note: This paper, originally published on 5/10/14, was replaced with a corrected/revised version on 5/19/14. If you downloaded the original PDF but are unable to access the revision, please contact SPIE Digital Library Customer Service for assistance.

  4. ConfocalCheck--a software tool for the automated monitoring of confocal microscope performance.

    Directory of Open Access Journals (Sweden)

    Keng Imm Hng

    Full Text Available Laser scanning confocal microscopy has become an invaluable tool in biomedical research but regular quality testing is vital to maintain the system's performance for diagnostic and research purposes. Although many methods have been devised over the years to characterise specific aspects of a confocal microscope like measuring the optical point spread function or the field illumination, only very few analysis tools are available. Our aim was to develop a comprehensive quality assurance framework ranging from image acquisition to automated analysis and documentation. We created standardised test data to assess the performance of the lasers, the objective lenses and other key components required for optimum confocal operation. The ConfocalCheck software presented here analyses the data fully automatically. It creates numerous visual outputs indicating potential issues requiring further investigation. By storing results in a web browser compatible file format the software greatly simplifies record keeping allowing the operator to quickly compare old and new data and to spot developing trends. We demonstrate that the systematic monitoring of confocal performance is essential in a core facility environment and how the quantitative measurements obtained can be used for the detailed characterisation of system components as well as for comparisons across multiple instruments.

  5. Axial resolution of confocal Raman microscopes : Gaussian beam theory and practice

    NARCIS (Netherlands)

    Grauw, C J De; Sijtsema, N M; Otto, C; Greve, J

    1997-01-01

    A straightforward and transparent model, based on Gaussian beam optics, for the axial ro resolution of a confocal microscope is presented, A confocal Raman microscope was used to determine the axial confocality in practice. The axial response of a thin planar object was measured for three different

  6. Axial Resolution of Confocal Raman Microscopes: Gaussian Beam Theory and Practice

    NARCIS (Netherlands)

    de Grauw, C.J.; de Grauw, C.J.; Sijtsema, N.M.; Otto, Cornelis; Greve, Jan

    1997-01-01

    A straightforward and transparent model, based on Gaussian beam optics, for the axial r0 resolution of a confocal microscope is presented. A confocal Raman microscope was used to determine the axial confocality in practice. The axial response of a thin planar object was measured for three different

  7. Adaptive optics in spinning disk microscopy: improved contrast and brightness by a simple and fast method.

    Science.gov (United States)

    Fraisier, V; Clouvel, G; Jasaitis, A; Dimitrov, A; Piolot, T; Salamero, J

    2015-09-01

    Multiconfocal microscopy gives a good compromise between fast imaging and reasonable resolution. However, the low intensity of live fluorescent emitters is a major limitation to this technique. Aberrations induced by the optical setup, especially the mismatch of the refractive index and the biological sample itself, distort the point spread function and further reduce the amount of detected photons. Altogether, this leads to impaired image quality, preventing accurate analysis of molecular processes in biological samples and imaging deep in the sample. The amount of detected fluorescence can be improved with adaptive optics. Here, we used a compact adaptive optics module (adaptive optics box for sectioning optical microscopy), which was specifically designed for spinning disk confocal microscopy. The module overcomes undesired anomalies by correcting for most of the aberrations in confocal imaging. Existing aberration detection methods require prior illumination, which bleaches the sample. To avoid multiple exposures of the sample, we established an experimental model describing the depth dependence of major aberrations. This model allows us to correct for those aberrations when performing a z-stack, gradually increasing the amplitude of the correction with depth. It does not require illumination of the sample for aberration detection, thus minimizing photobleaching and phototoxicity. With this model, we improved both signal-to-background ratio and image contrast. Here, we present comparative studies on a variety of biological samples. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  8. Planarization Profile Measurement Using A Confocal Scanning Laser Microscope

    Science.gov (United States)

    Smith, Ian R.; Bennett, Simon D.; Lindow, James T.; Monahan, Kevin

    1986-08-01

    The use of planarizing layers to improve the performance of photolithography for micron and submicron devices is being actively explored by a number of semiconductor companies. The usefulness of the procedure depends critically upon the degree to which residual surface undulations can be controlled. This paper describes how a confocal scanning optical microscope may be used to measure surface profiles of planarizing layers and discusses the factors which influence the accuracy of measurement. Experimental measurements, using a SiScan-I system, of resist and P.S.G. planarizing layers are presented, demonstrating a sensitivity to surface height changes of 50 nm. The technique may be improved upon by careful design of the microscope and selection of the imaging wavelength. These factors are discussed.

  9. Computerized Reconstruction of Pulpal Blood Vessels Examined under Confocal Microscope

    Directory of Open Access Journals (Sweden)

    Digka Anna

    2015-03-01

    Full Text Available The purpose of this study was the evaluation of 3 different histological methods for studying pulpal blood vessels in combination with 2 types of confocal microscope and computer assisted 3-dimensional reconstruction. 10 human, healthy, free of restorations or caries teeth that were extracted for orthodontic reasons were used. From these teeth, the pulp tissues of 5 were removed, fixed in formalin solution, dehydrated and embedded in paraffin. Serial cross sections 5μm thick were taken from 3 of the above mentioned pulpal tissues and stained with CD34 according to the immunohistochemical ABC technique, while the rest 2 were stained with CD34 and Cy5 by means of immunofluorescence after serial cross sectioning of 10μm. 5 of the 10 teeth were fixed, decalcified, serial cross sectioned (30μm thickness and stained with eosin. The physical sections were examined under 2 types of confocal laser microscope. Serial images were taken for each section, alignment of the images was followed and finally 3-dimensional reconstructions of the pulpal vessels were achieved.

  10. Live Imaging of Shoot Meristems on an Inverted Confocal Microscope Using an Objective Lens Inverter Attachment.

    Science.gov (United States)

    Nimchuk, Zachary L; Perdue, Tony D

    2017-01-01

    Live imaging of above ground meristems can lead to new insights in plant development not possible from static imaging of fixed tissue. The use of an upright confocal microscope offers several technical and biological advantages for live imaging floral or shoot meristems. However, many departments and core facilities possess only inverted confocal microscopes and lack the funding for an additional upright confocal microscope. Here we show that imaging of living apical meristems can be performed on existing inverted confocal microscopes with the use of an affordable and detachable InverterScope accessory.

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

    NARCIS (Netherlands)

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

    2017-01-01

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

  12. Upgrade of a Scanning Confocal Microscope to a Single-Beam Path STED Microscope.

    Directory of Open Access Journals (Sweden)

    André Klauss

    Full Text Available By overcoming the diffraction limit in light microscopy, super-resolution techniques, such as stimulated emission depletion (STED microscopy, are experiencing an increasing impact on life sciences. High costs and technically demanding setups, however, may still hinder a wider distribution of this innovation in biomedical research laboratories. As far-field microscopy is the most widely employed microscopy modality in the life sciences, upgrading already existing systems seems to be an attractive option for achieving diffraction-unlimited fluorescence microscopy in a cost-effective manner. Here, we demonstrate the successful upgrade of a commercial time-resolved confocal fluorescence microscope to an easy-to-align STED microscope in the single-beam path layout, previously proposed as "easy-STED", achieving lateral resolution < λ/10 corresponding to a five-fold improvement over a confocal modality. For this purpose, both the excitation and depletion laser beams pass through a commercially available segmented phase plate that creates the STED-doughnut light distribution in the focal plane, while leaving the excitation beam unaltered when implemented into the joint beam path. Diffraction-unlimited imaging of 20 nm-sized fluorescent beads as reference were achieved with the wavelength combination of 635 nm excitation and 766 nm depletion. To evaluate the STED performance in biological systems, we compared the popular phalloidin-coupled fluorescent dyes Atto647N and Abberior STAR635 by labeling F-actin filaments in vitro as well as through immunofluorescence recordings of microtubules in a complex epithelial tissue. Here, we applied a recently proposed deconvolution approach and showed that images obtained from time-gated pulsed STED microscopy may benefit concerning the signal-to-background ratio, from the joint deconvolution of sub-images with different spatial information which were extracted from offline time gating.

  13. 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...

  14. Confocal stereology: an efficient tool for measurement of microscopic structures

    Czech Academy of Sciences Publication Activity Database

    Kubínová, Lucie; Janáček, Jiří

    2015-01-01

    Roč. 360, č. 1 (2015), s. 13-28 ISSN 0302-766X R&D Projects: GA MŠk(CZ) LH13028 Institutional support: RVO:67985823 Keywords : 3-D images * confocal microscopy * geometrical characteristics * spatial probes * stereology Subject RIV: EA - Cell Biology Impact factor: 2.948, year: 2015

  15. Efficacy of oral exfoliative cytology in diabetes mellitus patients: a light microscopic and confocal microscopic study.

    Science.gov (United States)

    Gopal, Deepika; Malathi, N; Reddy, B Thirupathi

    2015-03-01

    Diabetes mellitus (DM) has become a global problem. By monitoring the health status of these individuals, diabetic complications can be prevented. We aimed to analyze alterations in the morphology and cytomorphometry of buccal epithelial cells of type 2 DM patients using oral exfoliative cytology technique and determine its importance in public health screening, diagnosis and monitoring of diabetes mellitus. The study was carried out in 100 type 2 DM patients and 30 healthy individuals. Smears were taken from the right buccal mucosa and stained by the Papanicolaou technique. Staining with Acridine orange was carried out to view qualitative changes with confocal laser scanning microscope (LSM-510 Meta). The cytomorphometry was evaluated using IMAGE PRO PLUS 5.5 software with Evolution LC camera. All findings were statistically analyzed. The results showed that with increase in fasting plasma glucose levels, there is significant increase in nuclear area, decrease in cytoplasmic area, and increase in nuclear cytoplasmic ratio (p diabetes and aid in monitoring of diabetes throughout the lifetime of the patient.

  16. Automated Assessment of Keratocyte Density in Stromal Images from the ConfoScan 4 Confocal Microscope

    Science.gov (United States)

    Bourne, William M.; Patel, Sanjay V.

    2010-01-01

    Purpose. To develop a program to determine cell densities in images from the ConfoScan 4 (Nidek, Inc., Freemont, CA) confocal microscope and compare the densities with those determined in images obtained by the Tandem Scanning confocal microscope (Tandem Scanning Corp., Reston, VA). Methods. A program was developed that used image-processing routines to identify stromal cell nuclei in images from the ConfoScan 4 confocal microscope. Cell selection parameters were set to match cell densities from the program with those determined manually in 15 normal corneas of 15 volunteers. The program was tested on scans from 16 other normal volunteers and 17 volunteers 3 years after LASIK. Cell densities were compared to densities determined by manual assessment and to those in scans by the Tandem Scanning confocal microscope in the same corneas. Results. The difference in cell density between the automatic and manual assessment was −539 ± 3005 cells/mm3 (mean ± SD, P = 0.11) in the 16 test corneas. Densities estimated from the ConfoScan 4 agreed with those from the Tandem Scanning confocal microscope in all regions of the stroma except in the anterior 10%, where the ConfoScan 4 indicated a 30% lower density. Conclusions. Differences in anterior stromal cell density between the ConfoScan 4 and the Tandem Scanning confocal microscope can be explained by the different optical designs. The lower spatial resolution of the ConfoScan 4 limits its ability to resolve thin layers. The adaptation of our earlier cell-counting program to the ConfoScan 4 provides a timesaving, objective, and reproducible means of determining stromal cell densities in images from the ConfoScan 4. PMID:19892869

  17. Demodex mites in acne rosacea: reflectance confocal microscopic study.

    Science.gov (United States)

    Turgut Erdemir, Asli; Gurel, Mehmet Salih; Koku Aksu, Ayse Esra; Falay, Tugba; Inan Yuksel, Esma; Sarikaya, Ebru

    2017-05-01

    Demodex mite density is emphasised in the aetiopathogenesis of acne rosacea. Reflectance confocal microscopy (RCM) has been shown to be a good method for determining demodex mite density. The objective was to determine demodex mite density using RCM in acne rosacea patients and compare them with controls. In all, 30 papulopustular rosacea (PPR) and 30 erythematotelangiectatic rosacea (ETR) totally 60 acne rosacea patients and 40 controls, were enrolled in the study. The right cheek was selected for imaging and RCM was used for scanning. Ten images of 1000 × 1000 μm (total 10 mm 2 ) area were scanned from adjacent areas. The numbers of follicles, infested follicles and mites were counted. The mean numbers of mites per follicle and infested follicles were calculated and compared in the patients and control groups. The mean number of mites was 44.30 ± 23.22 in PPR, 14.57 ± 15.86 in ETR and 3.55 ± 6.48 in the control group (P rosacea aetiopathogenesis. Demodex mite treatment may reduce the severity of the disease and slow its progressive nature. © 2016 The Australasian College of Dermatologists.

  18. Combined AFM and confocal fluorescence microscope for applications in bio-nanotechnology

    NARCIS (Netherlands)

    Kassies, R.; van der Werf, Kees; Lenferink, Aufrid T.M.; Hunter, C.N.; Olsen, J.D.; Subramaniam, Vinod; Otto, Cornelis

    2005-01-01

    We present a custom-designed atomic force fluorescence microscope (AFFM), which can perform simultaneous optical and topographic measurements with single molecule sensitivity throughout the whole visible to near-infrared spectral region. Integration of atomic force microscopy (AFM) and confocal

  19. A confocal optical microscope for detection of single impurities in a bulk crystal at cryogenic temperatures.

    Science.gov (United States)

    Karlsson, Jenny; Rippe, Lars; Kröll, Stefan

    2016-03-01

    A compact sample-scanning confocal optical microscope for detection of single impurities below the surface of a bulk crystal at cryogenic temperatures is described. The sample, lens, and scanners are mounted inside a helium bath cryostat and have a footprint of only 19 × 19 mm. Wide field imaging and confocal imaging using a Blu-ray lens immersed in liquid helium are demonstrated with excitation at 370 nm. A spatial resolution of 300 nm and a detection efficiency of 1.6% were achieved.

  20. Confocal microscope is able to detect calcium metabolic in neuronal infection by toxoplasma gondii

    Science.gov (United States)

    Sensusiati, A. D.; Priya, T. K. S.; Dachlan, Y. P.

    2017-05-01

    Calcium metabolism plays a very important role in neurons infected by Toxoplasma. Detection of change of calcium metabolism of neuron infected by Toxoplasma and Toxoplasma requires the calculation both quantitative and qualitative method. Confocal microscope has the ability to capture the wave of the fluorescent emission of the fluorescent dyes used in the measurement of cell calcium. The purpose of this study was to prove the difference in calcium changes between infected and uninfected neurons using confocal microscopy. Neuronal culture of human-skin-derived neural stem cell were divided into 6 groups, consisting 3 uninfected groups and 3 infected groups. Among the 3 groups were 2 hours, 24 hours and 48 hours. The neuron Toxoplasma gondii ratio was 1:5. Observation of intracellular calcium of neuron and tachyzoite, evidence of necrosis, apoptosis and the expression of Hsp 70 of neuron were examined by confocal microscope. The normality of the data was analysed by Kolmogorov-Smirnov Test, differentiation test was checked by t2 Test, and ANOVAs, for correlation test was done by Pearson Correlation Test. The calcium intensity of cytosolic neuron and T. gondii was significantly different from control groups (p<0.05). There was also significant correlation between calcium intensity with the evidence of necrosis and Hsp70 expression at 2 hours after infection. Apoptosis and necrosis were simultaneously shown with calcium contribution in this study. Confocal microscopy can be used to measure calcium changes in infected and uninfected neurons both in quantitatively and qualitatively.

  1. Multi-color miniature dual-axis confocal microscope for point-of-care pathology

    OpenAIRE

    Leigh, Steven Y.; Liu, Jonathan T.C.

    2012-01-01

    We present a miniature micro-electro-mechanical systems (MEMS)-based dual-axis confocal microscope capable of spatially co-registered fluorescence imaging at multiple wavelengths. This device has a 10-mm diameter scan head with a 2-mm diameter tip for convenient use during surgery to guide tumor resection. The microscope has an adjustable focal depth from 20 – 200 microns and is capable of imaging with an axial resolution of 9 microns and in-plane resolution of 4 microns over a field of view ...

  2. Confocal laser scanning microscope, Raman microscopy and Western blotting to evaluate inflammatory response after myocardial infarction.

    Science.gov (United States)

    Riezzo, Irene; Cantatore, Santina; DeCarlo, Dania; Fiore, Carmela; Neri, Margherita; Turillazzi, Emanuela; Fineschi, Vittorio

    2015-01-01

    Cardiac muscle necrosis is associated with inflammatory cascade that clears the infarct from dead cells and matrix debris, and then replaces the damaged tissue with scar, through three overlapping phases: the inflammatory phase, the proliferative phase and the maturation phase. Western blotting, laser confocal microscopy, Raman microscopy are valuable tools for studying the inflammatory response following myocardial infarction both humoral and cellular phase, allowing the identification and semiquantitative analysis of proteins produced during the inflammatory cascade activation and the topographical distribution and expression of proteins and cells involved in myocardial inflammation. Confocal laser scanning microscopy (CLSM) is a relatively new technique for microscopic imaging, that allows greater resolution, optical sectioning of the sample and three-dimensional reconstruction of the same sample. Western blotting used to detect the presence of a specific protein with antibody-antigen interaction in the midst of a complex protein mixture extracted from cells, produced semi-quantitative data quite easy to interpret. Confocal Raman microscopy combines the three-dimensional optical resolution of confocal microscopy and the sensitivity to molecular vibrations, which characterizes Raman spectroscopy. The combined use of western blotting and confocal microscope allows detecting the presence of proteins in the sample and trying to observe the exact location within the tissue, or the topographical distribution of the same. Once demonstrated the presence of proteins (cytokines, chemokines, etc.) is important to know the topographical distribution, obtaining in this way additional information regarding the extension of the inflammatory process in function of the time stayed from the time of myocardial infarction. These methods may be useful to study and define the expression of a wide range of inflammatory mediators at several different timepoints providing a more

  3. Design of an affordable fluorescence confocal laser scanning microscope for medical diagnostics

    Science.gov (United States)

    Bechtel, Christin; Knobbe, Jens; Grüger, Heinrich; Lakner, Hubert

    2012-12-01

    Confocal fluorescence microscopes are a promising imaging tool in medical diagnostics due to their capability to selectively survey cross-sections of individual layers from `thick' samples. Non-invasive depth resolved investigation of neoplastic skin disorders is one example among other applications. However these microscopes are at present uncommon in medical practice. This is due to their main application area in research. The instruments dealt with here are generally complex, stationary units and are accordingly cost-intensive. It is for this reason, that we have designed a robust and portable MEMS based confocal fluorescence microscope with a field of view of 0.6mm x 0.6mm. This has been made possible by the integration of a 2D micro scanner mirror developed at Fraunhofer IPMS. A variable acquisition depth of cross-sectional images of the fluorescence specimen is enabled by an integrated z-shifter. With the use of commercially available optics an optical demonstrator set up has been realized. To characterize and to demonstrate the ability of this system test measurements were performed. The resolution of the microscope is better than 228 lp/mm determined by 1951 USAF resolution test target. Images of various biological samples are presented and optical sectioning capabilities are shown. A comparison of the measured with the predicted system performance will be given.

  4. Design and analysis of a cross-type structured-illumination confocal microscope for high speed and high resolution

    International Nuclear Information System (INIS)

    Kim, Young-Duk; Ahn, MyoungKi; Kim, Taejoong; Gweon, DaeGab; Yoo, Hongki

    2012-01-01

    There have been many studies about a super resolution microscope for many years. A super resolution microscope can detect the physical phenomena or morphology of a biological sample more precisely than conventional microscopes. The structured-illumination microscope (SIM) is one of the technologies that demonstrate super resolution. However, the conventional SIM requires more time to obtain one resolution-enhanced image than other super resolution microscopes. More specifically, the conventional SIM uses three images with a 120° phase difference for each direction and three different directions are image-processed to make one resolution enhancement by increasing the optical transfer function in three directions. In this paper, we present a novel cross structured-illumination confocal microscope (CSICM) that takes the advantage of the technology of both SIM and the confocal microscope. The CSICM uses only two directions with three phase difference images, for a total of six images. By reducing the number of images that must be obtained, the total image acquisition time and image reconstruction time in obtaining the final output images can be decreased, and the confocal microscope provides axial information of the sample automatically. We demonstrate our method of cross illumination and evaluate the performance of the CSICM and compare it to the conventional SIM and the confocal microscope. (paper)

  5. A comparison of titanium alloy orthodontic wires for surface roughness using a confocal optical microscope

    OpenAIRE

    Hirokazu, Nakano; Akihide, Yoshida; Kazushi, Ogasawara; Akira, Sanjo; Shigeru, Tanaka; Takuya, Kamegai; Kazuro, Satoh; Hiroyuki, Miura; Department of Orthodontics, School of Dentistry, Iwate Medical University; Department of Orthodontics, School of Dentistry, Iwate Medical University; Department of Orthodontics, School of Dentistry, Iwate Medical University; Department of Orthodontics, School of Dentistry, Iwate Medical University; Department of Orthodontics, School of Dentistry, Iwate Medical University; Department of Orthodontics, School of Dentistry, Iwate Medical University; Department of Orthodontics, School of Dentistry, Iwate Medical University

    2001-01-01

    The purpose of this study was to clarify the surface roughness of 31 brands of titanium alloy orthodontic wires from 13 manufacturers using a confocal optical microscope. Cobalt-chrome and stainless steel wire were also examined as a reference of comparison. The following results were obtained ; (1) Mean Ra, as determined from the lengthway axis of titanium alloy wires, was 0.296μm, and that determined from the widthway axis was 0.440μm. The modulus of Ra was 0.368μm. (2) For titanium alloy o...

  6. Laser confocal microscope noise evaluation on injection compression moulded (ICM) transparent polymer Fresnel lenses

    DEFF Research Database (Denmark)

    Loaldi, D.; Calaon, Matteo; Quagliotti, Danilo

    The evaluation of an adequate and robust measuring strategy, for roughness assessment of polymer Fresnel lenses is put under assessment. An ‘on-sample’ measurement noise, is evaluated using a laser confocal microscope (OLYMPUS © Lext). Secondly, the lowest-noise roughness measuring procedure...... is performed by comparing ‘on-sample’ noise with the one calculated on an optical flat. Noise is investigated by means of established methods, i.e. subtraction and averaging methods. Afterwards, the lowest-noise analysis is structured following a 23 full factorial experimental planning, whose factors...

  7. 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.

  8. 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.

  9. Clinical features and confocal microscopic imaging characteristics of 466 cases with infectious keratitis

    Directory of Open Access Journals (Sweden)

    Hui Xiao

    2014-10-01

    Full Text Available AIM: To observe the role of confocal microscopy in infectious keratitis management. METHODS:Totally 466 patients(467 eyesdiagnosed as infectious keratitis from January 2010 to December 2013 were retrospectively studied. the corneas were examined early by in vivo confocal microscopy. The characteristics of their images and clinical features were studied and summarized.RESULTS:All patients were recorded, the average age was 54.4±13.0 years, in which 264 cases(56.7%were male, and 202 cases(43.3%were female. In the 466 patients, 190(40.8%were fungal keratitis, 148(31.8%were viral keratitis, 125(26.8%were bacterial keratitis and 3(0.6%were acanthamoeba keratitis. There were fungal hyphae in the images of fungal keratitis. Amebic cysts were found in acanthamoeba keratitis. CONCLUSION:Confocal microscope can help the early diagnose and treatment of infectious keratitis. It is a noninvasive imaging technique that provides high resolution images of ocular structures at a cellular level and infectious keratitis represents one of its most important clinical uses.

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

    International Nuclear Information System (INIS)

    Behan, G; Nellist, P D

    2008-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Karsten Rodenacker

    2011-05-01

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

  12. In Vivo Confocal Microscopic Evaluation of Corneal Endothelial Dysfunction Induced by Phacoemulcification in Rhesus Monkey Models.

    Science.gov (United States)

    Wu, Min; Hu, Zhu-lin; Sun, Xiao-mei; Dai, Jie-jie

    2016-02-01

    To observe the characteristic morphological changes of corneal endothelial dysfunction induced by phacoemulcification in rhesus monkey models under confocal microscope. The corneal endothelial dysfunction models were established by phacoemulcification power on the central corneal of 7 to 9 mm diameter in the right eyes of 4 rhesus monkeys (the modeling group). The left eyes of 4 rhesus monkeys were set as blank control group. The structural changes in different corneal layers were evaluated by slit lamp microscope and in vivo confocal microscope before surgery and 1, 2, 3, and 4 weeks after surgery. SPSS 19.0 software was applied to analyze data. Paired-t test was used to compare the number of nerve plexus in Bowman's layer and corneal endothelial cell density. Analysis of variance (ANOVA) was used to analyze corneal thickness. After phacoemulcification, the changes of cornea occurred gradually in the endothelial layer, stroma, Bowman's membrane, and basal epithelial layer. In the early stage, the interspace of corneal endothelial cells enlarged and few activated stromal cells were detected in the stroma. The cell morphology of stroma altered. The thickness of stroma increased. Two weeks after surgery, the nerve plexus in Bowman's layer decreased and edema of stroma and endothelial layer increased. Three weeks after surgery, the interspace of basal epithelial cells increased with a few Langerhans' cells infiltration and edema of stroma and endothelial layer increased. Four weeks after the surgery, a large amount of Langerhans' cells presented in basal epithelial layer. Only a few nerve lexus could be seen in Bowman's layer. The stroma and endothelial cells had severe edema. A large number of activated stromal cells could be found in stromal layer. Two weeks after the surgery, the number of nerve plexus in Bowman's layer (t=6.9192, P=0.002) and corneal endothelial cell density (t=7.8936, P<0.0001) in the modeling group were significantly lower than that in control

  13. Multi-color miniature dual-axis confocal microscope for point-of-care pathology.

    Science.gov (United States)

    Leigh, Steven Y; Liu, Jonathan T C

    2012-06-15

    We present a miniature microelectromechanical systems-based dual-axis confocal microscope capable of spatially coregistered fluorescence and reflectance imaging at multiple wavelengths. This device has a 10 mm diameter scan head with a 2 mm diameter tip for convenient use during surgery to guide tumor resection. The microscope has an adjustable focal depth of 20-200 micrometers and is capable of imaging with an axial resolution of 9 micrometers and in-plane resolution of 4 micrometers over a field of view of 450×450 micrometers. Simultaneous two-color imaging of individual optical sections is achieved by using a pair of grating-prism assemblies to compensate for chromatic dispersion in the 2 mm diameter gradient index relay lens at the distal tip of the device. Experimental measurements of the axial response of the microscope, as well as two-color images of a reflective bar target and fresh mouse brain tissues, demonstrate the performance of our device and its potential for multicolor in vivo optical sectioning microscopy.

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

    Science.gov (United States)

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

    2011-06-01

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

  15. 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.

  16. Polarization-preserving confocal microscope for optical experiments in a dilution refrigerator with high magnetic field.

    Science.gov (United States)

    Sladkov, Maksym; Bakker, M P; Chaubal, A U; Reuter, D; Wieck, A D; van der Wal, C H

    2011-04-01

    We present the design and operation of a fiber-based cryogenic confocal microscope. It is designed as a compact cold-finger that fits inside the bore of a superconducting magnet, and which is a modular unit that can be easily swapped between use in a dilution refrigerator and other cryostats. We aimed at application in quantum optical experiments with electron spins in semiconductors and the design has been optimized for driving with and detection of optical fields with well-defined polarizations. This was implemented with optical access via a polarization maintaining fiber together with Voigt geometry at the cold finger, which circumvents Faraday rotations in the optical components in high magnetic fields. Our unit is versatile for use in experiments that measure photoluminescence, reflection, or transmission, as we demonstrate with a quantum optical experiment with an ensemble of donor-bound electrons in a thin GaAs film. © 2011 American Institute of Physics

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

    Science.gov (United States)

    de Jonge, Niels [Oak Ridge, TN

    2010-08-17

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

  18. Comparison of three different sealer placement techniques: An In vitro confocal laser microscopic study

    Directory of Open Access Journals (Sweden)

    Avoy Kumar Dash

    2017-01-01

    Full Text Available Introduction: Three-dimensional obturation of the root canal system is the final objective of root canal therapy. Greater penetration of sealer in root dentine lesser will be the voids at the dentine–sealer interface. Hence, analysis of the dentin/sealer interface allows the determination of a filling technique which could obturate the root canals with least gaps and voids. Therefore, the aim of this study is to compare the depth and percentage of sealer penetration into root dentin using three different root canal sealer placement techniques under confocal laser scanning microscope. Materials and Methods: Thirty single-rooted teeth were selected and prepared. Adseal sealer (Meta Biomed, South Korea was mixed with Rhodamine B dye and applied using lentulo spiral (Dentsply Maillefer, USA as Group 1, bidirectional spiral (EZ-Fill– EDS, USA as Group 2, and ultrasonic endodontic tip (Sonofile– Dentsply Tulsa, USA as Group 3. Canals were then obturated with gutta-percha. The roots were sectioned at the 3 and 6-mm levels from the apical foramen and examined under confocal laser microscope. Results: Maximum mean depth and percentage of sealer penetration were observed for Group 1 and minimum for Group 3. Furthermore, statistical significant differences among Group 1 and Group 3 were found at 6-mm level and among Group 2 and Group 3 were found at 3-mm level (P < 0.05. Conclusion: The depth and percentage of sealer penetration of sealer are influenced by the type of placement technique and by the root canal level, with penetration decreasing apically. Lentulo spiral has shown better penetration of sealer than the bidirectional file and ultrasonics.

  19. Analysis of mitochondrial mechanical dynamics using a confocal fluorescence microscope with a bent optical fibre.

    Science.gov (United States)

    Li, Yongbo; Honda, Satoshi; Iwami, Kentaro; Ohta, Yoshihiro; Umeda, Norihiro

    2015-11-01

    The cells in the cardiovascular system are constantly subjected to mechanical forces created by blood flow and the beating heart. The effect of forces on cells has been extensively investigated, but their effect on cellular organelles such as mitochondria remains unclear. We examined the impact of nano-Newton forces on mitochondria using a bent optical fibre (BOF) with a flat-ended tip (diameter exceeding 2 μm) and a confocal fluorescence microscope. By indenting a single mitochondrion with the BOF tip, we found that the mitochondrial elastic modulus was proportional to the (-1/2) power of the mitochondrial radius in the 9.6-115 kPa range. We stained the mitochondria with a potential-metric dye (TMRE) and measured the changes in TMRE fluorescence intensity. We confirmed that more active mitochondria exhibit a higher frequency of repetitive transient depolarization. The same trend was observed at forces lower than 50 nN. We further showed that the depolarization frequency of mitochondria decreases under an extremely large force (nearly 100 nN). We conclude that mitochondrial function is affected by physical environmental factors, such as external forces at the nano-Newton level. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  20. A Generalization of Theory for Two-Dimensional Fluorescence Recovery after Photobleaching Applicable to Confocal Laser Scanning Microscopes

    Science.gov (United States)

    Kang, Minchul; Day, Charles A.; Drake, Kimberly; Kenworthy, Anne K.; DiBenedetto, Emmanuele

    2009-01-01

    Abstract Fluorescence recovery after photobleaching (FRAP) using confocal laser scanning microscopes (confocal FRAP) has become a valuable technique for studying the diffusion of biomolecules in cells. However, two-dimensional confocal FRAP sometimes yields results that vary with experimental setups, such as different bleaching protocols and bleaching spot sizes. In addition, when confocal FRAP is used to measure diffusion coefficients (D) for fast diffusing molecules, it often yields D-values that are one or two orders-of-magnitude smaller than that predicted theoretically or measured by alternative methods such as fluorescence correlation spectroscopy. Recently, it was demonstrated that this underestimation of D can be corrected by taking diffusion during photobleaching into consideration. However, there is currently no consensus on confocal FRAP theory, and no efforts have been made to unify theories on conventional and confocal FRAP. To this end, we generalized conventional FRAP theory to incorporate diffusion during photobleaching so that analysis by conventional FRAP theory for a circular region of interest is easily applicable to confocal FRAP. Finally, we demonstrate the accuracy of these new (to our knowledge) formulae by measuring D for soluble enhanced green fluorescent protein in aqueous glycerol solution and in the cytoplasm and nucleus of COS7 cells. PMID:19720039

  1. 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.

  2. 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)

  3. 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

    2014-02-24

    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.

  4. Surface profile measurement by using the integrated Linnik WLSI and confocal microscope system

    Science.gov (United States)

    Wang, Wei-Chung; Shen, Ming-Hsing; Hwang, Chi-Hung; Yu, Yun-Ting; Wang, Tzu-Fong

    2017-06-01

    The white-light scanning interferometer (WLSI) and confocal microscope (CM) are the two major optical inspection systems for measuring three-dimensional (3D) surface profile (SP) of micro specimens. Nevertheless, in practical applications, WLSI is more suitable for measuring smooth and low-slope surfaces. On the other hand, CM is more suitable for measuring uneven-reflective and low-reflective surfaces. As for aspect of surface profiles to be measured, the characteristics of WLSI and CM are also different. WLSI is generally used in semiconductor industry while CM is more popular in printed circuit board industry. In this paper, a self-assembled multi-function optical system was integrated to perform Linnik white-light scanning interferometer (Linnik WLSI) and CM. A connecting part composed of tubes, lenses and interferometer was used to conjunct finite and infinite optical systems for Linnik WLSI and CM in the self-assembled optical system. By adopting the flexibility of tubes and lenses, switching to perform two different optical measurements can be easily achieved. Furthermore, based on the shape from focus method with energy of Laplacian filter, the CM was developed to enhance the on focal information of each pixel so that the CM can provide all-in-focus image for performing the 3D SP measurement and analysis simultaneously. As for Linnik WLSI, eleven-step phase shifting algorithm was used to analyze vertical scanning signals and determine the 3D SP.

  5. 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.

  6. Super-resolution for everybody: An image processing workflow to obtain high-resolution images with a standard confocal microscope.

    Science.gov (United States)

    Lam, France; Cladière, Damien; Guillaume, Cyndélia; Wassmann, Katja; Bolte, Susanne

    2017-02-15

    In the presented work we aimed at improving confocal imaging to obtain highest possible resolution in thick biological samples, such as the mouse oocyte. We therefore developed an image processing workflow that allows improving the lateral and axial resolution of a standard confocal microscope. Our workflow comprises refractive index matching, the optimization of microscope hardware parameters and image restoration by deconvolution. We compare two different deconvolution algorithms, evaluate the necessity of denoising and establish the optimal image restoration procedure. We validate our workflow by imaging sub resolution fluorescent beads and measuring the maximum lateral and axial resolution of the confocal system. Subsequently, we apply the parameters to the imaging and data restoration of fluorescently labelled meiotic spindles of mouse oocytes. We measure a resolution increase of approximately 2-fold in the lateral and 3-fold in the axial direction throughout a depth of 60μm. This demonstrates that with our optimized workflow we reach a resolution that is comparable to 3D-SIM-imaging, but with better depth penetration for confocal images of beads and the biological sample. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Raster image correlation spectroscopy (RICS) for measuring fast protein dynamics and concentrations with a commercial laser scanning confocal microscope.

    Science.gov (United States)

    Brown, C M; Dalal, R B; Hebert, B; Digman, M A; Horwitz, A R; Gratton, E

    2008-01-01

    Raster image correlation spectroscopy (RICS) is a new and novel technique for measuring molecular dynamics and concentrations from fluorescence confocal images. The RICS technique extracts information about molecular dynamics and concentrations from images of living cells taken on commercial confocal systems. Here we develop guidelines for performing the RICS analysis on an analogue commercial laser scanning confocal microscope. Guidelines for typical instrument settings, image acquisition settings and analogue detector characterization are presented. Using appropriate instrument/acquisition parameters, diffusion coefficients and concentrations can be determined, even for highly dynamic dye molecules in solution. Standard curves presented herein demonstrate the ability to detect protein concentrations as low as approximately 2 nM. Additionally, cellular measurements give accurate values for the diffusion of paxillin-enhanced-green fluorescent protein (EGFP), an adhesion adaptor molecule, in the cytosol of the cell and also show slower paxillin dynamics near adhesions where paxillin interacts with immobile adhesion components. Methods are presented to account for bright immobile structures within the cell that dominate spatial correlation functions; allowing the extraction of fast protein dynamics within and near these structures. A running average algorithm is also presented to address slow cellular movement or movement of cellular features such as adhesions. Finally, methods to determine protein concentration in the presence of immobile structures within the cell are presented. A table is presented giving guidelines for instrument and imaging setting when performing RICS on the Olympus FV300 confocal and these guidelines are a starting point for performing the analysis on other commercial confocal systems.

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

    International Nuclear Information System (INIS)

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

    2010-01-01

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

  9. Optimizing the performance of dual-axis confocal microscopes via Monte-Carlo scattering simulations and diffraction theory.

    Science.gov (United States)

    Chen, Ye; Liu, Jonathan T C

    2013-06-01

    Dual-axis confocal (DAC) microscopy has been found to exhibit superior rejection of out-of-focus and multiply scattered background light compared to conventional single-axis confocal microscopy. DAC microscopes rely on the use of separated illumination and collection beam paths that focus and intersect at a single focal volume (voxel) within tissue. While it is generally recognized that the resolution and contrast of a DAC microscope depends on both the crossing angle of the DAC beams, 2θ, and the focusing numerical aperture of the individual beams, α, a detailed study to investigate these dependencies has not been performed. Contrast and resolution are considered as two main criteria to assess the performance of a point-scanned DAC microscope (DAC-PS) and a line-scanned DAC microscope (DAC-LS) as a function of θ and α. The contrast and resolution of these designs are evaluated by Monte-Carlo scattering simulations and diffraction theory calculations, respectively. These results can be used for guiding the optimal designs of DAC-PS and DAC-LS microscopes.

  10. Advanced spinning disk-TIRF microscopy for faster imaging of the cell interior and the plasma membrane.

    Science.gov (United States)

    Zobiak, Bernd; Failla, Antonio Virgilio

    2018-03-01

    Understanding the cellular processes that occur between the cytosol and the plasma membrane is an important task for biological research. Till now, however, it was not possible to combine fast and high-resolution imaging of both the isolated plasma membrane and the surrounding intracellular volume. Here, we demonstrate the combination of fast high-resolution spinning disk (SD) and total internal reflection fluorescence (TIRF) microscopy for specific imaging of the plasma membrane. A customised SD-TIRF microscope was used with specific design of the light paths that allowed, for the first time, live SD-TIRF experiments at high acquisition rates. A series of experiments is shown to demonstrate the feasibility and performance of our setup. © 2017 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.

  11. 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.

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

    Science.gov (United States)

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

    2016-01-01

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

  13. 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)

  14. Effects of Naphthalene on DNA and RNA quantity in Amoeba proteus by using confocal laser scanning microscope

    Directory of Open Access Journals (Sweden)

    Khwanmuni, J.

    2006-01-01

    Full Text Available Effects of Naphthalene which is a carcinogen on changes of DNA and RNA quantities were studied with acridine orange stained cells under a confocal laser scanning microscope. It was found that DNA and RNA in amoebae nucleus and cytoplasm, reared in 0 (control, 3 and 8.85 mg/l (24h-LD50 at 0 and 12 h. showed a statistically significant difference (p<0.05. The more naphthalene concentrations and larger incubation periods had greater effects on DNA and RNA decreases in amoebae nucleus and cytoplasm.

  15. Confocal scanning laser microscopic study of the RDX defect structure in deformed polymer-bonded explosives

    NARCIS (Netherlands)

    Heijden, A.E.D.M. van der; Bouma, R.H.B.

    2016-01-01

    The influence of an explosion-driven deformation on the defect structure in RDX crystals embedded in a polymer-bonded explosive was investigated by means of confocal scanning laser microscopy. The images were compared to the defect structure in the as-received RDX grades, embedded

  16. Dynamic experimentation on the confocal laser scanning microscope : application to soft-solid, composite food materials

    NARCIS (Netherlands)

    Plucknett, K.P.; Pomfret, S.J.; Normand, V.; Ferdinando, D.; Veerman, C.; Frith, W.J.; Norton, I.T.

    2001-01-01

    Confocal laser scanning microscopy (CLSM) is used to follow the dynamic structural evolution of several phase-separated mixed biopolymer gel composites. Two protein/polysaccharide mixed gel systems were examined: gelatin/maltodextrin and gelatin/agarose. These materials exhibit 'emulsion-like'

  17. 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

  18. Caustic meso-optical confocal microscope for vertical particle tracks. Proposal

    International Nuclear Information System (INIS)

    Soroko, L.M.

    1995-01-01

    The principal of the proposed caustic meso-optical microscope for vertical particle tracks in the nuclear photoemulsion is explained. The results of the experiments performed to illustrate the main features of this new meso-optical microscope are given. The proposed caustic meso-optical microscope for vertical particle tracks in the nuclear photoemulsion can be effectively used in the experimental investigation of such rare processes as ν μ - ν τ oscillations and of the Pb-Pb interactions. 2 refs., 7 figs

  19. Methods for compensation of the light attenuation with depth of images captured by a confocal microscope

    Czech Academy of Sciences Publication Activity Database

    Čapek, Martin; Janáček, Jiří; Kubínová, Lucie

    2006-01-01

    Roč. 69, č. 8 (2006), s. 624-635 ISSN 1059-910X R&D Projects: GA MŠk(CZ) LC06063; GA AV ČR(CZ) IAA100110502; GA AV ČR(CZ) IAA600110507; GA AV ČR(CZ) IAA500200510; GA AV ČR(CZ) KJB6011309; GA ČR(CZ) GA304/05/0153 Institutional research plan: CEZ:AV0Z50110509 Keywords : image enhancement * confocal microscopy * histogram warping Subject RIV: JC - Computer Hardware ; Software Impact factor: 1.680, year: 2006

  20. Optimization for imaging through scattering media for confocal microscopes with divided elliptical apertures.

    Science.gov (United States)

    Tang, Hengjie; Wu, Chenxue; Gong, Wei; Zheng, Yao; Zhu, Xinpei; Wang, Jiahao; Si, Ke

    2017-12-29

    We develop a confocal system equipped with optimal elliptical apertures to improve axial point spread function and signal-to-background ratio (SBR) for different detector sizes. By adjusting the parameters of the elliptical apertures, the axial half width at half-maximum can be reduced to 4.986 (described in optical coordinates) and SBR can be improved to 0.176. We evaluate this system with the 1951 USAF resolution test chart and the primary cultured neuron from SD rat stained by Map-2, and observe better imaging performance, which indicates the potential applications in biological science. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Semiautomated confocal imaging of fungal pathogenesis on plants: Microscopic analysis of macroscopic specimens.

    Science.gov (United States)

    Minker, Katharine R; Biedrzycki, Meredith L; Kolagunda, Abhishek; Rhein, Stephen; Perina, Fabiano J; Jacobs, Samuel S; Moore, Michael; Jamann, Tiffany M; Yang, Qin; Nelson, Rebecca; Balint-Kurti, Peter; Kambhamettu, Chandra; Wisser, Randall J; Caplan, Jeffrey L

    2018-02-01

    The study of phenotypic variation in plant pathogenesis provides fundamental information about the nature of disease resistance. Cellular mechanisms that alter pathogenesis can be elucidated with confocal microscopy; however, systematic phenotyping platforms-from sample processing to image analysis-to investigate this do not exist. We have developed a platform for 3D phenotyping of cellular features underlying variation in disease development by fluorescence-specific resolution of host and pathogen interactions across time (4D). A confocal microscopy phenotyping platform compatible with different maize-fungal pathosystems (fungi: Setosphaeria turcica, Cochliobolus heterostrophus, and Cercospora zeae-maydis) was developed. Protocols and techniques were standardized for sample fixation, optical clearing, species-specific combinatorial fluorescence staining, multisample imaging, and image processing for investigation at the macroscale. The sample preparation methods presented here overcome challenges to fluorescence imaging such as specimen thickness and topography as well as physiological characteristics of the samples such as tissue autofluorescence and presence of cuticle. The resulting imaging techniques provide interesting qualitative and quantitative information not possible with conventional light or electron 2D imaging. Microsc. Res. Tech., 81:141-152, 2018. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  2. Residual Endodontic Filling Material after Post Space Preparation: A Confocal Microscopic Study

    Directory of Open Access Journals (Sweden)

    Yu-Yao Teoh

    2017-11-01

    Full Text Available This laboratory study assessed removability of endodontic alkaline cements and resin sealers using coronal cross-sectional slices of roots with single canals. Materials were labelled with 0.1% (w/w sodium fluorescein prior to mixing so that confocal microscopy could be used to quantify material remaining on the walls of post spaces, to assess cleanliness. Roots of extracted teeth were prepared using rotary NiTi instruments then obturated using lateral condensation with gutta percha and epoxy resin sealers (AH-Plus™ or Zirmix™, or were filled by injecting mineral trioxide aggregate (MTA cement (GC Nex™ MTA or MTAmix™ or a hard-setting calcium hydroxide cement (Supercal™. Brown (#3 ParaPost™ drills were used at 600 rpm with a torque setting of 3 N cm−1 for 2 min to remove 5 mm of the root filling. Roots were embedded and coronal slices examined by confocal microscopy, with the perimeter of the drill channel divided into clean, unclean and non-accessible regions. The choice of material affects cleanliness, with MTA being the most difficult and calcium hydroxide cement the easiest to remove. With epoxy resin-based sealers, almost half of the accessible canal walls remained coated with remnants of sealer after post space preparation.

  3. Evaluation ofEnterococcus faecalisadhesion, penetration, and method to prevent the penetration ofEnterococcus faecalisinto root cementum: Confocal laser scanning microscope and scanning electron microscope analysis.

    Science.gov (United States)

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

    2016-01-01

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

  4. 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.

  5. A confocal microscopic study of solitary pulmonary neuroendocrine cells in human airway epithelium

    Directory of Open Access Journals (Sweden)

    Sparrow Malcolm P

    2005-10-01

    Full Text Available Abstract Background Pulmonary neuroendocrine cells (PNEC are specialized epithelial cells that are thought to play important roles in lung development and airway function. PNEC occur either singly or in clusters called neuroepithelial bodies. Our aim was to characterize the three dimensional morphology of PNEC, their distribution, and their relationship to the epithelial nerves in whole mounts of adult human bronchi using confocal microscopy. Methods Bronchi were resected from non-diseased portions of a lobe of human lung obtained from 8 thoracotomy patients (Table 1 undergoing surgery for the removal of lung tumors. Whole mounts were stained with antibodies to reveal all nerves (PGP 9.5, sensory nerves (calcitonin gene related peptide, CGRP, and PNEC (PGP 9.5, CGRP and gastrin releasing peptide, GRP. The analysis and rendition of the resulting three-dimensional data sets, including side-projections, was performed using NIH-Image software. Images were colorized and super-imposed using Adobe Photoshop. Results PNEC were abundant but not homogenously distributed within the epithelium, with densities ranging from 65/mm2 to denser patches of 250/mm2, depending on the individual wholemount. Rotation of 3-D images revealed a complex morphology; flask-like with the cell body near the basement membrane and a thick stem extending to the lumen. Long processes issued laterally from its base, some lumenal and others with feet-like processes. Calcitonin gene-related peptide (CGRP was present in about 20% of PNEC, mainly in the processes. CGRP-positive nerves were sparse, with some associated with the apical part of the PNEC. Conclusion Our 3D-data demonstrates that PNEC are numerous and exhibit a heterogeneous peptide content suggesting an active and diverse PNEC population.

  6. Visual and confocal microscopic interpretation of patch tests to benzethonium chloride and benzalkonium chloride.

    Science.gov (United States)

    Benjamin, Bohaty; Chris, Fricker; Salvador, González; Melissa, Gill; Susan, Nedorost

    2012-08-01

    Quaternary ammonium compounds (Quats), such as benzalkonium chloride (BAC) and benzethonium chloride (BEC), are widely used as antibacterial active ingredients and preservatives in personal care products, disinfectants, and ophthalmic preparations. BAC is known to be a marginal irritant when patch tested at 0.15% aq. Data on BEC are limited. To differentiate irritant from allergic patch test reactions to quaternary ammonium compounds. Eight subjects who were considered likely to react based on history of rash after exposure to disinfectants or a history of prior positive patch test to BAC were recruited, as well as two patients undergoing routine patch testing. BAC (0.15% aq), BAC (0.15% pet), BEC (0.05% aq), BEC (0.15% pet), BEC (0.15% aq), BEC (0.5% aq), sodium lauryl sulfate (2.0%), and deionized water were applied under Finn chambers for 48 h. Four days and 7 days after application, the sites were examined visually and then by in vivo reflectance confocal microscopy (RCM) which was interpreted by blinded experts. Two patients with definite allergic reactions according to visual patch test reads and RCM were clinically relevant. Cross-reaction between BEC and BAC was demonstrated in one patient. RCM imaging correlated well with clinical scoring and interpretation of patch test reactions in terms of irritancy vs. allergy for BEC and BAC. Relevant allergic reactions to quats occur in humans. Possible cross-reaction was noted to occur between BAC and BEC. RCM appears to be a useful tool in distinguishing between irritancy and sensitization during patch testing to BAC and BEC. Further study of prevalence and best test concentration and vehicle is needed. © 2011 John Wiley & Sons A/S.

  7. A confocal microscopic study of solitary pulmonary neuroendocrine cells in human airway epithelium.

    Science.gov (United States)

    Weichselbaum, Markus; Sparrow, Malcolm P; Hamilton, Elisha J; Thompson, Philip J; Knight, Darryl A

    2005-10-10

    Pulmonary neuroendocrine cells (PNEC) are specialized epithelial cells that are thought to play important roles in lung development and airway function. PNEC occur either singly or in clusters called neuroepithelial bodies. Our aim was to characterize the three dimensional morphology of PNEC, their distribution, and their relationship to the epithelial nerves in whole mounts of adult human bronchi using confocal microscopy. Bronchi were resected from non-diseased portions of a lobe of human lung obtained from 8 thoracotomy patients (Table 1) undergoing surgery for the removal of lung tumors. Whole mounts were stained with antibodies to reveal all nerves (PGP 9.5), sensory nerves (calcitonin gene related peptide, CGRP), and PNEC (PGP 9.5, CGRP and gastrin releasing peptide, GRP). The analysis and rendition of the resulting three-dimensional data sets, including side-projections, was performed using NIH-Image software. Images were colorized and super-imposed using Adobe Photoshop. PNEC were abundant but not homogenously distributed within the epithelium, with densities ranging from 65/mm2 to denser patches of 250/mm2, depending on the individual wholemount. Rotation of 3-D images revealed a complex morphology; flask-like with the cell body near the basement membrane and a thick stem extending to the lumen. Long processes issued laterally from its base, some lumenal and others with feet-like processes. Calcitonin gene-related peptide (CGRP) was present in about 20% of PNEC, mainly in the processes. CGRP-positive nerves were sparse, with some associated with the apical part of the PNEC. Our 3D-data demonstrates that PNEC are numerous and exhibit a heterogeneous peptide content suggesting an active and diverse PNEC population.

  8. In Vitro Assessment of Enamel Permeability in Primary Teeth with and without Early Childhood Caries Using Laser Scanning Confocal Microscope.

    Science.gov (United States)

    Niviethitha, Sabarinathan; Muthu, Murugan Satta; Kavitha, Swaminathan

    To compare the permeability of the enamel of primary teeth from individuals free of Early Childhood Caries (ECC) with that from individuals affected with ECC by assessment of dye penetration using Laser Scanning Confocal Microscope (LSCM). Experimental in vitro study. Exfoliated primary maxillary anterior teeth (n = 44) were collected and divided into two groups (n=22 per group): samples with ECC (Group 1) and without ECC (Group 2). The samples were immersed in Rhodamine B dye solution for 1 day, cut longitudinally into 3 sections, observed using LSCM. Dye penetration depths in the incisal, middle, cervical thirds and on labial, lingual surfaces were recorded. Data were analyzed by the Mann-Whitney test (α = 5%, p < .005). The overall mean penetration depth for group 1 (100.6 μm ± 58.48 μm) was significantly higher than that of group 2 (31.55 μm ± 23.40 μm, p < .000). Mean penetration depth in the incisal, middle, and cervical thirds and on the labial and lingual surfaces of group 1 also presented significantly higher scores than in group 2 (p < .005). There was significantly more dye penetration in the ECC group than in the non-ECC group. This could be related to a higher level of enamel permeability in teeth affected with ECC.

  9. Effect of the Nd:YAG laser on sealer penetration into root canal surfaces: a confocal microscope analysis.

    Science.gov (United States)

    Montero-Miralles, Paloma; Castillo-Oyagüe, Raquel; Sáez de la Fuente, Isabel; Lynch, Christopher D; Castillo-Dalí, Gabriel; Torres-Lagares, Daniel

    2014-06-01

    The objective of this in vitro study was to evaluate the use of the Neodymium:Yttrium-Aluminium-Garnet (Nd:YAG) laser as part of the root canal treatment on the penetration of sealer into dentinal tubules. Eighty extracted lower premolars were randomly assigned to two groups (n=40 each): Control group (CG), subjected to a conventional protocol of endodontic instrumentation and obturation; and Laser group (LG), in which Nd:YAG laser irradiations were combined with conventional preparation and obturation. Endodonted samples were sectioned at 3 and 5 mm from the apex and observed under a confocal scanning microscope (CLSM). The penetration depth into the dentinal tubules and the extension of the intracanal perimeter infiltrated by sealer were measured. The Student-Newman-Keuls test was run for between-group comparisons (α=.05). The depth of sealer penetration into dentinal tubules did not differ among groups. LG samples showed the significantly highest percentage of penetrated perimeter at 3 mm from the root apex. Within each group, the greatest depth of penetration (P=.0001), and the major percentage of penetrated perimeter (Plaser after instrumentation did not improve the depth of sealer penetration into the dentinal tubules. The laser enlarged the total penetrable perimeter near the apex. The Nd:YAG laser may be an appropriate complement in root canal treatment, as it enhances the sealer adaptation to the dentinal walls in the proximity of the apex. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. A sensitive and versatile laser scanning confocal optical microscope for single-molecule fluorescence at 77 K.

    Science.gov (United States)

    Hirschfeld, V; Hübner, C G

    2010-11-01

    We developed a cryostat suitable for a laser scanning confocal microscope which allows for a short working distance and thus the usage of an objective with a high numerical aperture ensuring high collection efficiency. The in situ preparation of a thin layer of amorphous water is realized in a part of the cryostat, a Dewar vessel, which is put onto a custom-made, liquid-nitrogen immersed spin-coater. First tests on the setup are performed on a perylenemonoimide/polymethyl methacrylate model system using a standard oil objective and a dry objective at ambient temperature as well as a dry objective at liquid nitrogen temperature (77 K). Fluorescence resonance energy transfer (FRET) measurements on doubly labeled, freeze-quenched polyproline chains show the applicability of the new method on biomolecules. The alternating laser excitation (ALEX) is modified to a line-scanning process (slow ALEX) to optimize the sorting of the labeled molecules. Photophysics and photochemistry at liquid nitrogen temperature are investigated.

  11. 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.

  12. Estimation of Nonconservative Aerodynamic Pressure Leading to Flutter of Spinning Disks

    DEFF Research Database (Denmark)

    Hansen, Morten Hartvig; Raman, A; Mote Jr., C.D.

    2001-01-01

    An experimental method for predicting the onset of #utter of a disk spinning in a #uid medium is proposed. The method is based on a description of the aerodynamic loading on the disk as a distributed viscous damping force rotating relative to the disk. This model can arise from two aeroelastic...... theories described herein. It is shown analytically and experimentally that the few parameters of this model may be extracted from frequency response functions of the spinning disk. Parameters for a steel disk in air (with a near vacuum experiment as reference) are estimated at increasing rotation speeds...

  13. Tailoring Cu Nanoparticle Catalyst for Methanol Synthesis Using the Spinning Disk Reactor

    Directory of Open Access Journals (Sweden)

    Christian Ahoba-Sam

    2018-01-01

    Full Text Available Cu nanoparticles are known to be very active for methanol (MeOH synthesis at relatively low temperatures, such that smaller particle sizes yield better MeOH productivity. We aimed to control Cu nanoparticle (NP size and size distribution for catalysing MeOH synthesis, by using the spinning disk reactor. The spinning disk reactor (SDR, which operates based on shear effect and plug flow in thin films, can be used to rapidly micro-mix reactants in order to control nucleation and particle growth for uniform particle size distribution. This could be achieved by varying both physical and chemical operation conditions in a precipitation reaction on the SDR. We have used the SDR for a Cu borohydride reduction to vary Cu NP size from 3 nm to about 55 nm. XRD and TEM characterization confirmed the presence of Cu2O and Cu crystallites when the samples were dried. This technique is readily scalable for Cu NP production by processing continuously over a longer duration than the small-scale tests. However, separation of the nanoparticles from solution posed a challenge as the suspension hardly settled. The Cu NPs produced were tested to be active catalyst for MeOH synthesis at low temperature and MeOH productivity increased with decreasing particle size.

  14. Tailoring Cu Nanoparticle Catalyst for Methanol Synthesis Using the Spinning Disk Reactor.

    Science.gov (United States)

    Ahoba-Sam, Christian; Boodhoo, Kamelia V K; Olsbye, Unni; Jens, Klaus-Joachim

    2018-01-17

    Cu nanoparticles are known to be very active for methanol (MeOH) synthesis at relatively low temperatures, such that smaller particle sizes yield better MeOH productivity. We aimed to control Cu nanoparticle (NP) size and size distribution for catalysing MeOH synthesis, by using the spinning disk reactor. The spinning disk reactor (SDR), which operates based on shear effect and plug flow in thin films, can be used to rapidly micro-mix reactants in order to control nucleation and particle growth for uniform particle size distribution. This could be achieved by varying both physical and chemical operation conditions in a precipitation reaction on the SDR. We have used the SDR for a Cu borohydride reduction to vary Cu NP size from 3 nm to about 55 nm. XRD and TEM characterization confirmed the presence of Cu₂O and Cu crystallites when the samples were dried. This technique is readily scalable for Cu NP production by processing continuously over a longer duration than the small-scale tests. However, separation of the nanoparticles from solution posed a challenge as the suspension hardly settled. The Cu NPs produced were tested to be active catalyst for MeOH synthesis at low temperature and MeOH productivity increased with decreasing particle size.

  15. Detection of adherence of Enterococcus faecalis in infected dentin of extracted human teeth using confocal laser scanning microscope: An In vitro Study

    Directory of Open Access Journals (Sweden)

    V Siddharth Nair

    2017-01-01

    Full Text Available Aim: The aim of this study is to detect in vitro adherence of Enterococcus faecalis to the infected dentinal tubules of human extracted teeth using confocal laser scanning microscope. Subjects and Methods: Roots from human premolar teeth (n = 40 were infected with E. faecalis strain the American Type Culture Collection 29212 in brain heart infusion for 21 days. After the experimental period, specimens were divided into two groups, Group A (n = 20, Group B (n = 20, and Group A specimens were stained with fluorescein diacetate dye for the detection of viability and adherence Group B were stained with acridine orange dye for detection of metabolic activity and adherence. Samples were washed, thoroughly sectioned and examined by confocal laser scanning microscopy. Computer-assisted determinants of fluorescence, bacterial viability, metabolic activity, and adherence were compared statistically. Results: E. faecalis was able to invade the dentinal tubules to a depth of 1–400 μm and adhere to 1–200 μm depth. Adherence (90% was significantly higher in 1–100 μm using fluorescein diacetate and acridine orange dye. Conclusion: Adherence of E. faecalis as evaluated by confocal laser scanning microscope was highest at the depth of 1–100 μm which may have an impact on the shaping and cleaning procedures on the root canal.

  16. A handheld MEMS-based line-scanned dual-axis confocal microscope for early cancer detection and surgical guidance (Conference Presentation)

    Science.gov (United States)

    Chen, Ye; Yin, Chengbo; Wei, Linpeng; Glaser, Adam K.; Abeytunge, Sanjee; Peterson, Gary; Mandella, Michael J.; Sanai, Nader; Rajadhyaksha, Milind; Liu, Jonathan T.

    2017-02-01

    Considerable efforts have been recently undertaken to develop miniature optical-sectioning microscopes for in vivo microendoscopy and point-of-care pathology. These devices enable in vivo interrogation of disease as a real-time and noninvasive alternative to gold-standard histopathology, and therefore could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Regardless of the specific modality, various trade-offs in size, speed, field of view, resolution, contrast, and sensitivity are necessary to optimize a device for a particular application. Here, a miniature MEMS-based line-scanned dual-axis confocal (LS-DAC) microscope, with a 12-mm diameter distal tip, has been developed for point-of-care pathology. The dual-axis architecture has demonstrated superior rejection of out-of-focus and multiply scattered photons compared to a conventional single-axis confocal configuration. The use of line scanning enables fast frame rates (≥15 frames/sec), which mitigates motion artifacts of a handheld device during clinical use. We have developed a method to actively align the illumination and collection beams in this miniature LS-DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo application, enables the device to achieve an axial and lateral resolution of 2.0 and 1.1 microns, respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate that this high-speed LS-DAC microscope can achieve high-contrast imaging of fluorescently labeled tissues with sufficient sensitivity for applications such as oral cancer detection and guiding brain-tumor resections.

  17. Handheld tunable focus confocal microscope utilizing a double-clad fiber coupler for in vivo imaging of oral epithelium

    Science.gov (United States)

    Olsovsky, Cory; Hinsdale, Taylor; Cuenca, Rodrigo; Cheng, Yi-Shing Lisa; Wright, John M.; Rees, Terry D.; Jo, Javier A.; Maitland, Kristen C.

    2017-05-01

    A reflectance confocal endomicroscope with double-clad fiber coupler and electrically tunable focus lens is applied to imaging of the oral mucosa. The instrument is designed to be lightweight and robust for clinical use. The tunable lens allows axial scanning through >250 μm in the epithelium when the probe tip is placed in contact with tissue. Images are acquired at 6.6 frames per second with a field of view diameter up to 850 μm. In vivo imaging of a wide range of normal sites in the oral cavity demonstrates the accessibility of the handheld probe. In vivo imaging of clinical lesions diagnosed as inflammation and dysplasia illustrates the ability of reflectance confocal endomicroscopy to image cellular changes associated with pathology.

  18. 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.

  19. Reinnervation of the tibialis anterior following sciatic nerve crush injury: a confocal microscopic study in transgenic mice.

    Science.gov (United States)

    Magill, Christina K; Tong, Alice; Kawamura, David; Hayashi, Ayato; Hunter, Daniel A; Parsadanian, Alexander; Mackinnon, Susan E; Myckatyn, Terence M

    2007-09-01

    Transgenic mice whose axons and Schwann cells express fluorescent chromophores enable new imaging techniques and augment concepts in developmental neurobiology. The utility of these tools in the study of traumatic nerve injury depends on employing nerve models that are amenable to microsurgical manipulation and gauging functional recovery. Motor recovery from sciatic nerve crush injury is studied here by evaluating motor endplates of the tibialis anterior muscle, which is innervated by the deep peroneal branch of the sciatic nerve. Following sciatic nerve crush, the deep surface of the tibialis anterior muscle is examined using whole mount confocal microscopy, and reinnervation is characterized by imaging fluorescent axons or Schwann cells (SCs). One week following sciatic crush injury, 100% of motor endplates are denervated with partial reinnervation at 2 weeks, hyperinnervation at 3 and 4 weeks, and restoration of a 1:1 axon to motor endplate relationship 6 weeks after injury. Walking track analysis reveals progressive recovery of sciatic nerve function by 6 weeks. SCs reveal reduced S100 expression within 2 weeks of denervation, correlating with regression to a more immature phenotype. Reinnervation of SCs restores S100 expression and a fully differentiated phenotype. Following denervation, there is altered morphology of circumscribed terminal Schwann cells demonstrating extensive process formation between adjacent motor endplates. The thin, uniformly innervated tibialis anterior muscle is well suited for studying motor reinnervation following sciatic nerve injury. Confocal microscopy may be performed coincident with other techniques of assessing nerve regeneration and functional recovery.

  20. Molecular characterization and confocal laser scanning microscopic study of Pygidiopsis macrostomum (Trematoda: Heterophyidae) parasites of guppies Poecilia vivipara.

    Science.gov (United States)

    Borges, J N; Costa, V S; Mantovani, C; Barros, E; Santos, E G N; Mafra, C L; Santos, C P

    2017-02-01

    Pygidiopsis macrostomum and Ascocotyle (Phagicola) pindoramensis (Digenea: Heterophyidae) parasitize guppies as intermediate hosts and, respectively, fish-eating mammals or birds as definitive hosts. Heterophyids have zoonotic potential, and molecular studies associated with morphological and ecological aspects have helped to clarify their taxonomy and phylogeny. Poecilia vivipara naturally parasitized by metacercariae of both species (100% prevalence) exhibit no external signs of parasitism. In this work, four new sequences of P. macrostomum (18S rDNA, 28S rDNA and ITS2 rDNA) and one new sequence of A. (P.) pindoramensis (mtDNA cox-1) are presented. Phylogeny reconstructions linked P. macrostomum to other heterophyids, but the separation of the Heterophyidae and Opisthorchiidae remains unclear. Additionally, we used indirect immunocytochemistry and the phalloidin-fluorescence techniques allied with confocal laser scanning microscopy to describe muscular and neuronal structures of P. macrostomum. A complex arrangement of muscular fibres is associated with the tegument, suckers, gut and reproductive system. Radial fibres around the ventral sucker are thick, branched and extend to the body wall. High-resolution confocal imaging revealed a typical digenean muscular arrangement and important heterophyid morphological traits. These data will support future control measures to reduce the parasitism in guppies reared in fish farming systems, especially for aquarium and experimental purposes. © 2016 John Wiley & Sons Ltd.

  1. Fluorescence correlation spectroscopy, Raster image correlation spectroscopy and Number & Brightness on a commercial confocal laser scanning microscope with analog detectors (Nikon C1)

    Science.gov (United States)

    Moens, Pierre D.J.; Gratton, Enrico; Salvemini, Iyrri L.

    2010-01-01

    Fluorescence correlation spectroscopy (FCS) was developed in 1972 by Magde, Elson and Webb (Magde et al., 1972). Photon counting detectors and avalanche photodiodes have become standards in FCS to the point that there is a widespread belief that these detectors are essential to perform FCS experiments, despite the fact that FCS was developed using analog detectors. Spatial and temporal intensity fluctuation correlations using analog detection on a commercial Olympus Fluoview 300 microscope has been reported by Brown et al. (2008). However, each analog instrument has its own idiosyncrasies that need to be understood before using the instrument for FCS. In this work we explore the capabilities of the Nikon C1, a low cost confocal microscope, to obtain single point FCS, Raster-scan Image Correlation Spectroscopy (RICS) and Number & Brightness data both in solution and incorporated into the membrane of Giant Unilamellar Vesicles (GUVs). We show that it is possible to obtain dynamic information about fluorescent molecules from single point FCS, RICS and Number & Brightness using the Nikon C1. We highlighted the fact that care should be taken in selecting the acquisition parameters in order to avoid possible artifacts due to the detector noise. However, due to relatively large errors in determining the distribution of digital levels for a given microscope setting, the system is probably only adequate for determining relative brightness within the same image. PMID:20734406

  2. Fluorescence correlation spectroscopy, raster image correlation spectroscopy, and number and brightness on a commercial confocal laser scanning microscope with analog detectors (Nikon C1).

    Science.gov (United States)

    Moens, Pierre D J; Gratton, Enrico; Salvemini, Iyrri L

    2011-04-01

    Fluorescence correlation spectroscopy (FCS) was developed in 1972 by Magde, Elson and Webb. Photon counting detectors and avalanche photodiodes have become standards in FCS to the point that there is a widespread belief that these detectors are essential to perform FCS experiments, despite the fact that FCS was developed using analog detectors. Spatial and temporal intensity fluctuation correlations using analog detection on a commercial Olympus Fluoview 300 microscope have been reported by Brown et al. (2008). However, each analog instrument has its own idiosyncrasies that need to be understood before using the instrument for FCS. In this work, we explore the capabilities of the Nikon C1, a low-cost confocal microscope, to obtain single point FCS, Raster-scan image correlation spectroscopy (RICS), and Number and Brightness data both in solution and incorporated into the membrane of giant unilamellar vesicles. We show that it is possible to obtain dynamic information about fluorescent molecules from single point FCS, RICS, and Number and Brightness using the Nikon C1. We highlighted the fact that care should be taken in selecting the acquisition parameters to avoid possible artifacts due to the detector noise. However, due to relatively large errors in determining the distribution of digital levels for a given microscope setting, the system is probably only adequate for determining relative brightness within the same image. Copyright © 2010 Wiley-Liss, Inc.

  3. Aberration correction during real time in vivo imaging of bone marrow with sensorless adaptive optics confocal microscope.

    Science.gov (United States)

    Wang, Zhibin; Wei, Dan; Wei, Ling; He, Yi; Shi, Guohua; Wei, Xunbin; Zhang, Yudong

    2014-08-01

    We have demonstrated adaptive correction of specimen-induced aberration during in vivo imaging of mouse bone marrow vasculature with confocal fluorescence microscopy. Adaptive optics system was completed with wavefront sensorless correction scheme based on stochastic parallel gradient descent algorithm. Using image sharpness as the optimization metric, aberration correction was performed based upon Zernike polynomial modes. The experimental results revealed the improved signal and resolution leading to a substantially enhanced image contrast with aberration correction. The image quality of vessels at 38- and 75-μm depth increased three times and two times, respectively. The corrections allowed us to detect clearer bone marrow vasculature structures at greater contrast and improve the signal-to-noise ratio.

  4. Upgrading the GSI beamline microscope with a confocal fluorescence lifetime scanner to monitor charged particle induced chromatin decondensation in living cells

    Energy Technology Data Exchange (ETDEWEB)

    Abdollahi, Elham; Taucher-Scholz, Gisela [Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse 1, 64291 Darmstadt (Germany); Durante, Marco [Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse 1, 64291 Darmstadt (Germany); Institute for Condensed Matter Physics, Darmstadt University of Technology, 64289 Darmstadt (Germany); Jakob, Burkhard, E-mail: B.Jakob@gsi.de [Department of Biophysics, GSI Helmholtz Center for Heavy Ion Research, Planckstrasse 1, 64291 Darmstadt (Germany)

    2015-12-15

    We report the upgrade of the GSI beamline microscope coupled to the linear accelerator UNILAC by a confocal FLIM scanner utilizing time correlated single photon counting technique (TCSPC). The system can now be used to address the radiation induced chromatin decondensation in more detail and with higher sensitivity compared to intensity based methods. This decondensation of heterochromatic areas is one of the early DNA damage responses observed after charged particle irradiation and might facilitate the further processing of the induced lesions. We describe here the establishment of different DNA dyes as chromatin compaction probes usable for quantification of the DNA condensation status in living cells utilizing lifetime imaging. In addition, we find an evidence of heterochromatic chromatin decondensation in ion irradiated murine chromocenters detected after subsequent fixation using FLIM measurements.

  5. 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.

  6. Confocal laser scanning and electron microscopical studies on osmoregulatory epithelia in the branchial cavity of the lobster homarus gammarus

    Science.gov (United States)

    Haond; Flik; Charmantier

    1998-06-01

    The adult lobster Homarus gammarus is a weak hyper-regulator at low salinity. The objective of this study was to locate the ion-transporting tissues in the branchial chamber of this species, using electron microscopy and confocal laser scanning microscopy with a fluorescent vital stain for mitochondria, DASPMI, which is widely used to locate mitochondria-rich cells in ion-transporting epithelia of fish. A thick mitochondria-rich epithelium is present on the inner side of the branchiostegite and over the entire surface of the epipodites. Ultrastructural observations confirm that this tissue has features typical of an ion-transporting epithelium. When the lobster is transferred to low salinity, these epithelia undergo marked ultrastructural changes, such as an increase in thickness related to the development of basolateral infoldings, the appearance of numerous vesicles and an increase in height of the apical microvilli. In the gills, the branchial filaments are lined by a thin and poorly differentiated epithelium, containing numerous mitochondria; no significant ultrastructural changes were observed in the gills of animals acclimated to low salinity. In summary, in H. gammarus, no evidence of osmoregulatory structures was found in the gills. Differentiated ion-transporting epithelia are present in the branchial cavity, on the inner side of the branchiostegite and on the epipodites; these organs are probably involved in osmoregulation.

  7. Axially-confined in vivo single-cell labeling by primed conversion using blue and red lasers with conventional confocal microscopes.

    Science.gov (United States)

    Taniguchi, Atsushi; Kimura, Yukiko; Mori, Ikue; Nonaka, Shigenori; Higashijima, Shin-Ichi

    2017-12-01

    Green-to-red photoconvertible fluorescent proteins have been found to undergo efficient photoconversion by a new method termed primed conversion that uses dual wave-length illumination with blue and red/near-infrared light. By modifying a confocal laser-scanning microscope (CLSM) such that two laser beams only meet at the focal plane, confined photoconversion at the axial dimension has been achieved. The necessity of this custom modification to the CLSM, however, has precluded the wide-spread use of this method. Here, we investigated whether spatially-restricted primed conversion could be achieved with CLSM without any hardware modification. We found that the primed conversion of Dendra2 using a conventional CLSM with two visible lasers (473 nm and 635 nm) and a high NA objective lens (NA, 1.30) resulted in dramatic restriction of photoconversion volume: half-width half-maximum for the axial dimension was below 5 μm, which is comparable to the outcome of the original method that used the microscope modification. As a proof of this method's effectiveness, we used this technique in living zebrafish embryos and succeeded in revealing the complex anatomy of individual neurons packed between neighboring cells. Because unmodified CLSMs are widely available, this method can be widely applicable for labeling cells with single-cell resolution. © 2017 The Authors Development, Growth & Differentiation published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Developmental Biologists.

  8. 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.

  9. In vivo confocal microscopic investigation of the cornea after autologous implantation of lenticules obtained through small incision lenticule extraction for treatment of hyperopia.

    Science.gov (United States)

    Li, Meiyan; Li, Meng; Sun, Ling; Han, Tian; Ding, Lan; Xiang, Jun; Zhou, Xingtao

    2018-01-01

    To investigate re-innervation in the implanted lenticule, as well as changes to the cornea, after correcting hyperopia with an autologous implantation of a lenticule obtained through small incision lenticule extraction (SMILE). This study retrospectively analysed re-innervation in the implanted lenticule, as well as microscopic morphological changes in the corneal architecture of the recipient cornea in five patients (with myopia in one eye and hyperopia in the contralateral eye) who received SMILE in the myopic eye and femtosecond laser in situ keratomileusis (FS-LASIK) combined with lenticule implantation in the contralateral hyperopic eye. Re-innervation in the implanted lenticule, as well as microscopic morphological changes in corneal architecture, were evaluated by in vivo confocal microscopy. One patient was examined at postoperative week six, two were examined at postoperative month two, one was examined at postoperative month nine, and one was examined at postoperative month 12. Partial subbasal nerve fibre regeneration was detected in two patients (Case 4 who was examined at postoperative month nine and Case 5 who was examined at postoperative month 12). Regenerated and branched nerve fibres were detected in the implanted lenticule in Case 5 who was examined at postoperative month 12. Both the anterior and posterior interfaces showed an absence or decrease of keratocytes and the presence of small particles of various brightnesses. Keratocytes in the implanted lenticule presented abnormal morphology in size and shape after surgery in all treated eyes, but showed partial morphological recovery in two patients (Case 4 who was examined at postoperative month nine and Case 5 who was examined at postoperative month 12). These preliminary findings suggest that nerve fibres will regenerate into the implanted lenticule after autologous lenticule implantation. Keratocytes in lenticules demonstrated a gradual return to a normal morphology. © 2017 Optometry

  10. Series Solution for Steady Three-Dimensional Flow due to Spraying on Inclined Spinning Disk by Homotopy Perturbation Method

    Directory of Open Access Journals (Sweden)

    Saeed Dinarvand

    2012-01-01

    Full Text Available The steady three-dimensional flow of condensation or spraying on inclined spinning disk is studied analytically. The governing nonlinear equations and their associated boundary conditions are transformed into the system of nonlinear ordinary differential equations. The series solution of the problem is obtained by utilizing the homotopy perturbation method (HPM. The velocity and temperature profiles are shown and the influence of Prandtl number on the heat transfer and Nusselt number is discussed in detail. The validity of our solutions is verified by the numerical results. Unlike free surface flows on an incline, this through flow is highly affected by the spray rate and the rotation of the disk.

  11. Confocal nanoscanning, bead picking (CONA): PickoScreen microscopes for automated and quantitative screening of one-bead one-compound libraries.

    Science.gov (United States)

    Hintersteiner, Martin; Buehler, Christof; Uhl, Volker; Schmied, Mario; Müller, Jürgen; Kottig, Karsten; Auer, Manfred

    2009-01-01

    Solid phase combinatorial chemistry provides fast and cost-effective access to large bead based libraries with compound numbers easily exceeding tens of thousands of compounds. Incubating one-bead one-compound library beads with fluorescently labeled target proteins and identifying and isolating the beads which contain a bound target protein, potentially represents one of the most powerful generic primary high throughput screening formats. On-bead screening (OBS) based on this detection principle can be carried out with limited automation. Often hit bead detection, i.e. recognizing beads with a fluorescently labeled protein bound to the compound on the bead, relies on eye-inspection under a wide-field microscope. Using low resolution detection techniques, the identification of hit beads and their ranking is limited by a low fluorescence signal intensity and varying levels of the library beads' autofluorescence. To exploit the full potential of an OBS process, reliable methods for both automated quantitative detection of hit beads and their subsequent isolation are needed. In a joint collaborative effort with Evotec Technologies (now Perkin-Elmer Cellular Technologies Germany GmbH), we have built two confocal bead scanner and picker platforms PS02 and a high-speed variant PS04 dedicated to automated high resolution OBS. The PS0X instruments combine fully automated confocal large area scanning of a bead monolayer at the bottom of standard MTP plates with semiautomated isolation of individual hit beads via hydraulic-driven picker capillaries. The quantification of fluorescence intensities with high spatial resolution in the equatorial plane of each bead allows for a reliable discrimination between entirely bright autofluorescent beads and real hit beads which exhibit an increased fluorescence signal at the outer few micrometers of the bead. The achieved screening speed of up to 200,000 bead assayed in less than 7 h and the picking time of approximately 1 bead

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

    Science.gov (United States)

    Sonomura, Takahiro; Furuta, Takahiro; Nakatani, Ikuko; Yamamoto, Yo; Honma, Satoru; Kaneko, Takeshi

    2014-11-01

    Ten years have passed since a serial block-face scanning electron microscopy (SBF-SEM) method was developed [1]. In this innovative method, samples were automatically sectioned with an ultramicrotome placed inside a scanning electron microscope column, and the block surfaces were imaged one after another by SEM to capture back-scattered electrons. The contrast-inverted images obtained by the SBF-SEM were very similar to those acquired using conventional TEM. SFB-SEM has made easy to acquire image stacks of the transmission electron microscopy (TEM) in the mesoscale, which is taken with the confocal laser-scanning microcopy(CF-LSM).Furthermore, serial-section SEM has been combined with the focused ion beam (FIB) milling method [2]. FIB-incorporated SEM (FIB-SEM) has enabled the acquisition of three-dimensional images with a higher z-axis resolution com- pared to ultramicrotome-equipped SEM.We tried immunocytochemistry for FIB-SEM and correlated this immunoreactivity with that in CF-LSM. Dendrites of neurons in the rat neostriatum were visualized using a recombinant viral vector. Moreover, the thalamostriatal afferent terminals were immunolabeled with Cy5 fluorescence for vesicular glutamate transporter 2 (VGluT2). After detection of the sites of terminals apposed to the dendrites by using CF-LSM, GFP and VGluT2 immunoreactivities were further developed for EM by using immunogold/silver enhancement and immunoperoxidase/diaminobenzidine (DAB) methods, respectively.We showed that conventional immuno-cytochemical staining for TEM was applicable to FIB-SEM. Furthermore, several synaptic contacts, which were thought to exist on the basis of CF-LSM findings, were confirmed with FIB-SEM, revealing the usefulness of the combined method of CF-LSM and FIB-SEM. © The Author 2014. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. Confocal microscopy

    Indian Academy of Sciences (India)

    This is elucidated by time-resolved confocal microscopy. Keywords. Porphyrin; micro-rod; anisotropy; exciton coupling; confocal microscopy. 1. Introduction. Supra-molecular assemblies of porphyrin play a central role in light harvesting during photosynthesis.1 10 In such a system, the absorbed photon shuttles between dif-.

  14. 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...

  15. Conjugation of both on-axis and off-axis light in Nipkow disk confocal microscope to increase availability of incoherent light source.

    Science.gov (United States)

    Saito, Kenta; Arai, Yoshiyuki; Zhang, Jize; Kobayashi, Kentaro; Tani, Tomomi; Nagai, Takeharu

    2011-01-01

    Laser-scanning confocal microscopy has been employed for exploring structures at subcellular, cellular and tissue level in three dimensions. To acquire the confocal image, a coherent light source, such as laser, is generally required in conventional single-point scanning microscopy. The illuminating beam must be focused onto a small spot with diffraction-limited size, and this determines the spatial resolution of the microscopy system. In contrast, multipoint scanning confocal microscopy using a Nipkow disk enables the use of an incoherent light source. We previously demonstrated successful application of a 100 W mercury arc lamp as a light source for the Yokogawa confocal scanner unit in which a microlens array was coupled with a Nipkow disk to focus the collimated incident light onto a pinhole (Saito et al., Cell Struct. Funct., 33: 133-141, 2008). However, transmission efficiency of incident light through the pinhole array was low because off-axis light, the major component of the incident light, was blocked by the non-aperture area of the disk. To improve transmission efficiency, we propose an optical system in which off-axis light is able to be transmitted through pinholes surrounding the pinhole located on the optical axis of the collimator lens. This optical system facilitates the use of not only the on-axis but also the off-axis light such that the available incident light is considerably improved. As a result, we apply the proposed system to high-speed confocal and multicolor imaging both with a satisfactory signal-to-noise ratio.

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

    Directory of Open Access Journals (Sweden)

    I. A. Volkov

    2014-01-01

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

  17. 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.

  18. Effect of EndoActivator and Er,Cr:YSGG laser activation of Qmix, as final endodontic irrigant, on sealer penetration: A Confocal microscopic study.

    Science.gov (United States)

    Chaudhry, Sarika; Yadav, Sudha; Talwar, Sangeeta; Verma, Mahesh

    2017-02-01

    Through chemomechanical debridement of the root canal is a primary requisite for successful endodontic therapy. Thus the aim of this study was to evaluate the effects of using QmiX alone, QmiX with EndoActivator and QmiX with Er,Cr:YSGG laser for final irrigation on sealer penetration into the dentinal tubules. 75 extracted human mandibular premolar teeth were treated with sodium hypochlorite (NaOCl) irrigation. The samples were divided into 5 groups according to the final irrigation solution used: (1) 17% EDTA and 2.5% NaOCl, (2) QmiX (3) QmiX with Er,Cr:YSGG laser and (4) QmiX with EndoActivator (5) 2.5%NaOCl. All teeth were obturated using cold lateral condensation technique with gutta percha and AH 26 sealer (Dentsply; DeTrey,Konstanz, Germany) labeled with Rhodamine B dye. The teeth were sectioned at distances of 2 and 5 from root apex. Total percentage and maximum depth of sealer penetration were measured using confocal laser scanning microscopy. Results of one way Anova analysis showed that there was a significant difference in the percentage and depth of sealer penetration among all groups at 3 and 5 mm level sections ( P laser activated group. Greater depth of sealer penetration was recorded at 5mm as compared to 3mm in all the groups. Activation of QMix using EndoActivator and Er,Cr:YSGG laser enhanced the sealer penetration at apical and middle third. Thus Er,Cr:YSGG laser and EndoActivator may act as an appropriate adjunct during chemomechanical preparation of the root canal. Key words: EndoActivator, Er,Cr:YSGG laser, Qmix, confocal microscopy, sealer penetration.

  19. Molecular confocal laser endomicroscopy

    DEFF Research Database (Denmark)

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

    2014-01-01

    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...... as future challenges of molecular CLE in gastrointestinal diseases....

  20. Light Microscopy Research Group (LMRG): International Test Results for Objective Lens Quality, Resolution, Spectral Accuracy and Spectral Separation for Confocal Laser Scanning Microscopes (CLSM)

    Science.gov (United States)

    Thibault, Marc; Bayles, Carol J.; Eason, Brady; Girard, Anne-Marie; Jinadasa, Tushare; Opansky, Cynthia; Schulz, Katherine; Brown, Claire M.; Cole, Richard W.

    2013-01-01

    As part of an ongoing effort to increase image reproducibility and fidelity in addition to improving cross-instrument consistency we have developed four separate instrument quality tests to augment the ones previously reported. 1) Objective lens quality 2) Resolution 3) Spectral accuracy of the wavelength information from spectral detectors 4) Spectral Separation tested the accuracy and quality of un-mixing algorithms. To ascertain the usefulness of these tests as well as to determine the current ““state”” of microscopes in use, test specimens and detailed protocols were made available worldwide free-of-charge. 55 laboratories located in 18 countries provided data. Objective lens quality: good with most issues arising from user errors or stage/focus drift, approximately 10% of lenses had aberrations. Resolution: within an average of approximately 25% of theoretical values. Spectral accuracy: excellent, even for low resolution systems. Spectral un-mixing: good, poor data collection was the main cause of low quality data.

  1. In vivo confocal Raman microscopic determination of depth profiles of the stratum corneum lipid organization influenced by application of various oils.

    Science.gov (United States)

    Choe, ChunSik; Schleusener, Johannes; Lademann, Jürgen; Darvin, Maxim E

    2017-08-01

    The intercellular lipids (ICL) of stratum corneum (SC) play an important role in maintaining the skin barrier function. The lateral and lamellar packing order of ICL in SC is not homogenous, but rather depth-dependent. This study aimed to analyze the influence of the topically applied mineral-derived (paraffin and petrolatum) and plant-derived (almond oil and jojoba oil) oils on the depth-dependent ICL profile ordering of the SC in vivo. Confocal Raman microscopy (CRM), a unique tool to analyze the depth profile of the ICL structure non-invasively, is employed to investigate the interaction between oils and human SC in vivo. The results show that the response of SC to oils' permeation varies in the depths. All oils remain in the upper layers of the SC (0-20% of SC thickness) and show predominated differences of ICL ordering from intact skin. In these depths, skin treated with plant-derived oils shows more disordered lateral and lamellar packing order of ICL than intact skin (poils do not influence the lateral packing order of SC ICL (p>0.1), except plant-derived oils at the depth 30% of SC thickness. In the deeper layers of the SC (60-100% of SC thickness), no difference between ICL lateral packing order of the oil-treated and intact skin can be observed, except that at the depths of 70-90% of the SC thickness, where slight changes with more disorder states are measured for plant-derived oil treated skin (poil types remain in the superficial layers of the SC (0-20% of the SC thickness). Skin treated with mineral- and plant-derived oils shows significantly higher disordered lateral and lamellar packing order of ICL in these layers of the SC compared to intact skin. Plant-derived oils significantly changed the ICL ordering in the depths of 30% and 70-90% of the SC thickness, which is likely due to the penetration of free fatty acids in the deeper layers of the SC. Copyright © 2017 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights

  2. 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

  3. 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

  4. HeNe laser (633 nm)-coupled confocal microscope allows simulating magnetic resonance imaging/computed tomography scan of the brain and eye: a noninvasive optical approach applicable to small laboratory animals.

    Science.gov (United States)

    Zheng, Ping-Pin; Romme, Edwin; van der Spek, Peter J; Dirven, Clemens M F; Willemsen, Rob; Kros, Johan M

    2011-06-01

    Magnetic resonance imaging (MRI) and computed tomography (CT) are noninvasive medical imaging techniques used for the detailed visualization of internal organs of the human body. Because CT uses X-rays for imaging, there is a risk of radiation exposure. In contrast, MRI uses radiowaves and magnetic fields for imaging; thus, there are no reported biological hazards. However, neither MRI nor CT is suitable as a noninvasive imaging tool applicable in small laboratory animals such as zebrafish embryos or larvae. The recently established micro-CT scanner is only suitable for scanning adult fish and a staining procedure is required for imaging. In addition, CT-based scanning is generally more suitable for skeletal imaging but not for visualization of soft tissues because of its lower contrast. In this study, we evaluated whether 633 nm HeNe laser-coupled confocal microscope allows simulating MRI/CT scan and imaging soft tissues such as brain and eye in zebrafish embryos/larvae. We show that the 633 nm HeNe laser can penetrate well into intact brain and eye of zebrafish. It represents a noninvasive imaging method with high resolution while not requiring contrast agents, enabling the detection of differential signals from normal and pathological organs such as brain and eye.

  5. Confocal microendoscopy with chromatic sectioning

    Science.gov (United States)

    Lane, Pierre M.; Elliott, Robert P.; MacAulay, Calum E.

    2003-07-01

    Placing a spatial light modulator, such as the Texas Instruments Digital Micromirror Device (DMD), in the light path of a microscope enables a variety of novel applications. One application enables reflectance in vivo confocal imaging of cells and tissue structure through a fiber-optic image guide. While multi-wavelength reflectance confocal microendoscopy with optical sectioning is a requirement for a clinically useful device, some form of axial scanning is also necessary. This is readily achieved using a multi-element lens system with some form of mechanical translation, however, this generally results in large probes and high cost. These limitations can be overcome using a two-element GRIN lens system in which the traditionally undesirable chromatic aberration of such a system can be exploited to allow for color-encoded optical sectioning. In our system a wavelength encoding range of 200 nm permits a sectioning range of 40 μm from the tip of the probe into the tissue.

  6. Low-power, Confocal Imaging of Protein Localization in Living Cells (7214-150), Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — The proposed technology genetically labels intracellular structures and visualizes protein interactions in living cells using a compact, confocal microscope with...

  7. 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.

  8. 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.

  9. Practical aspects of quantitative confocal microscopy.

    Science.gov (United States)

    Murray, John M

    2013-01-01

    Confocal microscopes are in principle well suited for quantitative imaging. The 3D fluorophore distribution in a specimen is transformed by the microscope optics and detector into the 2D intensity distribution of a digital image by a linear operation, a convolution. If multiple 2D images of the specimen at different focal planes are obtained, then the original 3D distribution in the specimen can be reconstructed. This reconstruction is a low-pass spatially filtered representation of the original, but quantitatively preserves relative fluorophore concentrations, with of course some limitations on accuracy and precision due to aberrations and noise. Given appropriate calibration, absolute fluorophore concentrations are accessible. A few simple guidelines are given for setting up confocal microscopes and checking their performance. With a little care, the images collected should be suitable for most types of quantitative analysis. Copyright © 2013 Elsevier Inc. All rights reserved.

  10. 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.

  11. 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....... Confocal microscopes are held by many research groups, and a number of methods for qualitative and quantitative imaging of the matrix have emerged in recent years. This review provides an overview and a critical discussion of techniques used to visualize different matrix compounds, to determine...... the concentration of solutes and the diffusive properties of the biofilm matrix....

  12. Recent advances in confocal microscopy for studying drug delivery to the eye: concepts and pharmaceutical applications.

    Science.gov (United States)

    Furrer, Pascal; Gurny, Robert

    2010-01-01

    Since its seminal introduction 50years ago, confocal microscopy has been applied in numerous fields in life sciences. This review presents the different key elements of confocal microscopes, in particular scanning techniques, light sources and especially laser sources are described in this review. Furthermore, an overview of the different image processing systems coupled with confocal microscopy is provided. The chapter closes with the applications of confocal microscopy in drug delivery to the eye. Copyright 2009 Elsevier B.V. All rights reserved.

  13. Real-Time Live Confocal Fluorescence Microscopy as a New Tool for Assessing Platelet Vitality.

    Science.gov (United States)

    Hermann, Martin; Nussbaumer, Oliver; Knöfler, Ralf; Hengster, Paul; Nussbaumer, Walter; Streif, Werner

    2010-01-01

    BACKGROUND: Assessment of platelet vitality is important for patients presenting with inherited or acquired disorders of platelet function and for quality assessment of platelet concentrates. METHODS: Herein we combined live stains with intra-vital confocal fluorescence microscopy in order to obtain an imaging method that allows fast and accurate assessment of platelet vitality. Three fluorescent dyes, FITC-coupled wheat germ agglutinin (WGA), tetramethylrhodamine methyl ester perchlorate (TMRM) and acetoxymethylester (Rhod-2), were used to assess platelet morphology, mitochondrial activity and intra-platelet calcium levels. Microscopy was performed with a microlens-enhanced Nipkow spinning disk-based system allowing live confocal imaging. RESULTS: Comparison of ten samples of donor platelets collected before apheresis and platelets collected on days 5 and 7 of storage showed an increase in the percentage of Rhod-2-positive platelets from 3.6 to 47 and finally to 71%. Mitochondrial potential was demonstrated in 95.4% of donor platelets and in 92.5% of platelets stored for 7 days. CONCLUSION: Such fast and accurate visualization of known key parameters of platelet function could be of relevance for studies addressing the quality of platelets after storage and additional manipulation, such as pathogen inactivation, as well as for the analysis of inherited platelet function disorders.

  14. 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.

  15. Diagnosis of Acanthamoeba keratitis in vivo with confocal microscopy.

    Science.gov (United States)

    Winchester, K; Mathers, W D; Sutphin, J E; Daley, T E

    1995-01-01

    We present eight cases of Acanthamoeba keratitis. In each case; the Acanthamoeba organisms were visualized in the epithelium and anterior stroma using tandem scanning confocal microscopy. The organisms were highly reflective, ovoid, and were 10-25 microns in diameter. The Acanthamoeba organisms in the human corneas were identical in size and shape to Acanthamoeba organisms on an agar plate visualized with the same confocal microscope. Confocal microscopy is a useful method for identifying Acanthamoeba organisms in vivo within the corneal epithelium and anterior stroma.

  16. Confocal Raman microscope mapping as a tool to describe different mineral and organic phases at high spatial resolution within marine biogenic carbonates: case study on Nerita undata (Gastropoda, Neritopsina

    Directory of Open Access Journals (Sweden)

    J. Nouet

    2011-12-01

    Full Text Available Marine biogenic carbonates formed by invertebrates (e.g. corals and mollusks represent complex composites of one or more mineral phases and organic molecules. This complexity ranges from the macroscopic structures observed with the naked eye down to sub micrometric structures only revealed by micro analytical techniques. Understanding to what extent and how organisms can control the formation of these structures requires that the mineral and organic phases can be identified and their spatial distribution related. Here we demonstrate the capability of confocal Raman microscopy applied to cross sections of a shell of Nerita undata to describe the distribution of calcite and aragonite including their crystallographic orientation with high lateral resolution (~300 nm. Moreover, spatial distribution of functional groups of organic compounds can be simultaneously acquired, allowing to specifically relate them to the observed microstructures. The data presented in this case study highlights the possible new contributions of this method to the description of modalities of Nerita undata shell formation, and what could be expected of its application to other marine biogenic carbonates. Localization of areas of interest would also allow further investigations using more localized methods, such as TEM that would provide complementary information on the relation between organic molecules and crystal lattice.

  17. Confocal Raman microscope mapping as a tool to describe different mineral and organic phases at high spatial resolution within marine biogenic carbonates: case study on Nerita undata (Gastropoda, Neritopsina)

    Science.gov (United States)

    Nehrke, G.; Nouet, J.

    2011-12-01

    Marine biogenic carbonates formed by invertebrates (e.g. corals and mollusks) represent complex composites of one or more mineral phases and organic molecules. This complexity ranges from the macroscopic structures observed with the naked eye down to sub micrometric structures only revealed by micro analytical techniques. Understanding to what extent and how organisms can control the formation of these structures requires that the mineral and organic phases can be identified and their spatial distribution related. Here we demonstrate the capability of confocal Raman microscopy applied to cross sections of a shell of Nerita undata to describe the distribution of calcite and aragonite including their crystallographic orientation with high lateral resolution (~300 nm). Moreover, spatial distribution of functional groups of organic compounds can be simultaneously acquired, allowing to specifically relate them to the observed microstructures. The data presented in this case study highlights the possible new contributions of this method to the description of modalities of Nerita undata shell formation, and what could be expected of its application to other marine biogenic carbonates. Localization of areas of interest would also allow further investigations using more localized methods, such as TEM that would provide complementary information on the relation between organic molecules and crystal lattice.

  18. Video-rate scanning confocal microscopy and microendoscopy.

    Science.gov (United States)

    Nichols, Alexander J; Evans, Conor L

    2011-10-20

    Confocal microscopy has become an invaluable tool in biology and the biomedical sciences, enabling rapid, high-sensitivity, and high-resolution optical sectioning of complex systems. Confocal microscopy is routinely used, for example, to study specific cellular targets, monitor dynamics in living cells, and visualize the three dimensional evolution of entire organisms. Extensions of confocal imaging systems, such as confocal microendoscopes, allow for high-resolution imaging in vivo and are currently being applied to disease imaging and diagnosis in clinical settings. Confocal microscopy provides three-dimensional resolution by creating so-called "optical sections" using straightforward geometrical optics. In a standard wide-field microscope, fluorescence generated from a sample is collected by an objective lens and relayed directly to a detector. While acceptable for imaging thin samples, thick samples become blurred by fluorescence generated above and below the objective focal plane. In contrast, confocal microscopy enables virtual, optical sectioning of samples, rejecting out-of-focus light to build high resolution three-dimensional representations of samples. Confocal microscopes achieve this feat by using a confocal aperture in the detection beam path. The fluorescence collected from a sample by the objective is relayed back through the scanning mirrors and through the primary dichroic mirror, a mirror carefully selected to reflect shorter wavelengths such as the laser excitation beam while passing the longer, Stokes-shifted fluorescence emission. This long-wavelength fluorescence signal is then passed to a pair of lenses on either side of a pinhole that is positioned at a plane exactly conjugate with the focal plane of the objective lens. Photons collected from the focal volume of the object are collimated by the objective lens and are focused by the confocal lenses through the pinhole. Fluorescence generated above or below the focal plane will therefore not

  19. A novel confocal line scanning sensor

    Science.gov (United States)

    Chanbai, Sirichanok; Wiora, Georg; Weber, Mark; Roth, Hubert

    2009-05-01

    Optical methods, including confocal microscopes, are widely used for measurements of surface topography. The knowledge of surface morphology and roughness parameters is crucial for many applications, i.e. in industrial and automotive environment, in tribology, wear and functionality prediction. However, confocal microscopy has a limited field of view. A time consuming stitching process is required for extending to long profile lines measurement. Therefore, in this paper we present a novel concept of a Confocal Line Scanning Sensor (CLSS) to cover theoretically infinite profile lengths. The new technique is proposed with no moving parts required for axial scanning, and it has a simpler setup than those of Chromatic Confocal Sensor (CCS). The idea is to produce a stack of focal points on an inclined plane covering a certain axial measurement range. Therefore, by scanning the stack of focal points in lateral direction we can realize a long profile line. By doing that we expect to achieve shorter scanning time, while providing high lateral and axial resolution by using a true confocal principle. A long profile line of a few ten millimeters with a lateral resolution in sub-micrometer range and an axial resolution in tens of nanometers can be expected. Moreover, this concept is easily extensible to an areal measurement. Among other key components, a new design of the pinhole mask has been developed. We design it to produce an inclined focal line with optimum optical parameters. Optimization of the pinhole design fulfills two objectives; minimizing its size by allowing optimal reflected-light intensity, and minimizing crosstalk between nearby pinholes. Further detail of the pinhole design is beyond a scope of this paper. In this paper an overview of the new concept is presented, accompanied by validation of first experimental results.

  20. 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. ...

  1. 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

  2. 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

  3. Scanning differential polarization microscope: Its use to image linear and circular differential scattering

    International Nuclear Information System (INIS)

    Mickols, W.; Maestre, M.F.

    1988-01-01

    A differential polarization microscope that couples the sensitivity of single-beam measurement of circular dichroism and circular differential scattering with the simultaneous measurement of linear dichroism and linear differential scattering has been developed. The microscope uses a scanning microscope stage and single-point illumination to give the very shallow depth of field found in confocal microscopy. This microscope can operate in the confocal mode as well as in the near confocal condition that can allow one to program the coherence and spatial resolution of the microscope. This microscope has been used to study the change in the structure of chromatin during the development of sperm in Drosophila

  4. Confocal Raman microscopy for identification of bacterial species in biofilms

    Science.gov (United States)

    Beier, Brooke D.; Quivey, Robert G.; Berger, Andrew J.

    2011-03-01

    Implemented through a confocal microscope, Raman spectroscopy has been used to distinguish between biofilm samples of two common oral bacteria species, Streptococcus sanguinis and mutans, which are associated with healthy and cariogenic plaque, respectively. Biofilms of these species are studied as a model of dental plaque. A prediction model has been calibrated and validated using pure biofilms. This model has been used to identify the species of transferred and dehydrated samples (much like a plaque scraping) as well as hydrated biofilms in situ. Preliminary results of confocal Raman mapping of species in an intact two-species biofilm will be shown.

  5. A MEANS FOR CALIBRATING A MICROSCOPE, A METHOD OF PREPARING THE MEANS AND A METHOD OF CALIBRATING A MICROSCOPE

    DEFF Research Database (Denmark)

    2005-01-01

    A means or artefact for calibrating the height/depth or Z axis of a microscope, such as a confocal microscope, an interference microscope or a Scanning Electron Microscope. The artefact comprises a number of tapering or pie-shaped, parallel surfaces each extending from a central axis, whereby all...... surfaces and heights are visible independently of the magnification of the microscope. Thus, the full height range may be calibrated and the linearity thereof checked independently of the magnification....

  6. A confocal laser scanning microscopic study on thermoresponsive ...

    Indian Academy of Sciences (India)

    These suspensions can be utilized for making binary colloidal alloys. Acknowledgements. Authors acknowledges Dr Baldev Raj, Dr C S Sundar and Dr A K Arora, Materials. Science Group, IGCAR for support and encouragement. First author acknowledges. UGC-DAE-CSR, Kolkata for financial support. References. [1] J Li ...

  7. A confocal laser scanning microscopic study on thermoresponsive

    Indian Academy of Sciences (India)

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

  8. A confocal laser scanning microscopic study on thermoresponsive ...

    Indian Academy of Sciences (India)

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

  9. A confocal laser scanning microscopic study on thermoresponsive ...

    Indian Academy of Sciences (India)

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

  10. Physical chemistry in a single live cell: confocal microscopy.

    Science.gov (United States)

    Amin, Md Asif; Nandi, Somen; Mondal, Prasenjit; Mahata, Tanushree; Ghosh, Surajit; Bhattacharyya, Kankan

    2017-05-24

    A live cell is a complex, yet extremely important container. Understanding the dynamics in a selected intracellular component is a challenging task. We have recently made significant progress in this direction using a confocal microscope as a tool. The smallest size of the focused spot in a confocal microscope is ∼0.2 μm (200 nm). This is nearly one hundred times smaller than the size of a live cell. Thus, one can selectively study different intracellular components/organelles in a live cell. In this paper, we discuss how one can image different intracellular components/organelles, record fluorescence spectra and decay at different locations, ascertain local polarity and viscosity, and monitor the dynamics of solvation, proton transfer, red-ox and other phenomena at specified locations/organelles inside a cell. We will highlight how this knowledge enriched us in differentiating between cancer and non-cancer cells, 3D tumor spheroids and towards drug delivery.

  11. A Study on 3-Dimensional Surface Measurement using Confocal Principle

    International Nuclear Information System (INIS)

    Kang, Young June; Song, Dae Ho; You, Weon Jae

    2001-01-01

    In modern industry, the accuracy and the sulfate-finish requirements for machined parts have been becoming ever more stringent. In addition, the measurement and understanding of surface topography is rapidly attracting the attention of the physicist and chemist as well as the engineer. Optical measuring method is used in vibration measurement, crack and defect detection with the advent of opto-mechatronics, and it is expected to play an important role in surface topography. In this study, the principle of confocal microscope is described, and the advanced 3-D surface measuring system that has better performance than the traditional confocal microscope is developed. Suitable fixtures arc developed and integrated with the computer system for generating 3-D surface and form data. Software for data acquisition and analysis of various parameters in surface geometrical features has been developed

  12. 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

  13. Submicron confocal Raman imaging of holograms in multicomponent photopolymers

    Science.gov (United States)

    Kagan, C. R.; Harris, T. D.; Harris, A. L.; Schilling, M. L.

    1998-04-01

    We report submicron chemical imaging of optical holograms in multicomponent photopolymers using a confocal scanning Raman microscope. The microscope is sensitive to the submicron, ˜1% concentration variations of the polymeric components that form refractive index modulations (Δn) responsible for hologram diffraction. Concentration variations are established by both small molecule diffusion and polymer matrix swelling during hologram writing. Both density and composition variations contribute to Δn. These measurements demonstrate that submicron Raman microscopy is applicable to multicomponent organic, inorganic, and hybrid materials as a route to correlate materials chemistry/morphology with their physical properties.

  14. Confocal laser endomicroscopy

    DEFF Research Database (Denmark)

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

    2016-01-01

    and to examine whether these are risk factors for relapse. PATIENTS AND METHODS: This was a single-center prospective feasibility study of CLE imaging in patients with Crohn's disease. CLE imaging was performed in the terminal ileum and four colorectal sites, and was correlated with histopathology...... a 12-month follow-up period, ileal fluorescein leakage and microerosions were significant risk factors for relapse in the subgroup of patients in remission (log rank (P = 0.009 and (P = 0.007, respectively) as well as in the entire group of patients with Crohn's disease (log rank (P = 0.006 and (P = 0.......01, respectively). Inter- and intraobserver reproducibility was almost perfect (κ > 0.80) or substantial (κ > 0.60) for the majority of CLE parameters. CONCLUSIONS: CLE can identify reproducible microscopic changes in the terminal ileum that are risk factors for relapse in patients with otherwise inactive Crohn...

  15. 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 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 resolution. In

  16. Proper alignment of the microscope.

    Science.gov (United States)

    Rottenfusser, Rudi

    2013-01-01

    The light microscope is merely the first element of an imaging system in a research facility. Such a system may include high-speed and/or high-resolution image acquisition capabilities, confocal technologies, and super-resolution methods of various types. Yet more than ever, the proverb "garbage in-garbage out" remains a fact. Image manipulations may be used to conceal a suboptimal microscope setup, but an artifact-free image can only be obtained when the microscope is optimally aligned, both mechanically and optically. Something else is often overlooked in the quest to get the best image out of the microscope: Proper sample preparation! The microscope optics can only do its job when its design criteria are matched to the specimen or vice versa. The specimen itself, the mounting medium, the cover slip, and the type of immersion medium (if applicable) are all part of the total optical makeup. To get the best results out of a microscope, understanding the functions of all of its variable components is important. Only then one knows how to optimize these components for the intended application. Different approaches might be chosen to discuss all of the microscope's components. We decided to follow the light path which starts with the light source and ends at the camera or the eyepieces. To add more transparency to this sequence, the section up to the microscope stage was called the "Illuminating Section", to be followed by the "Imaging Section" which starts with the microscope objective. After understanding the various components, we can start "working with the microscope." To get the best resolution and contrast from the microscope, the practice of "Koehler Illumination" should be understood and followed by every serious microscopist. Step-by-step instructions as well as illustrations of the beam path in an upright and inverted microscope are included in this chapter. A few practical considerations are listed in Section 3. Copyright © 2013 Elsevier Inc. All rights

  17. Microscope basics.

    Science.gov (United States)

    Sluder, Greenfield; Nordberg, Joshua J

    2013-01-01

    This chapter provides information on how microscopes work and discusses some of the microscope issues to be considered in using a video camera on the microscope. There are two types of microscopes in use today for research in cell biology-the older finite tube-length (typically 160mm mechanical tube length) microscopes and the infinity optics microscopes that are now produced. The objective lens forms a magnified, real image of the specimen at a specific distance from the objective known as the intermediate image plane. All objectives are designed to be used with the specimen at a defined distance from the front lens element of the objective (the working distance) so that the image formed is located at a specific location in the microscope. Infinity optics microscopes differ from the finite tube-length microscopes in that the objectives are designed to project the image of the specimen to infinity and do not, on their own, form a real image of the specimen. Three types of objectives are in common use today-plan achromats, plan apochromats, and plan fluorite lenses. The concept of mounting video cameras on the microscope is also presented in the chapter. Copyright © 2003 Elsevier Inc. All rights reserved.

  18. Microwave Microscope

    Data.gov (United States)

    Federal Laboratory Consortium — FUNCTION: Makes ultra-high-resolution field measurements. The Microwave Microscope (MWM) has been used in support of several NRL experimental programs involving sea...

  19. 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.

  20. Development of confocal X-ray fluorescence (XRF) microscopy at the Cornell high energy synchrotron source

    International Nuclear Information System (INIS)

    Woll, A.R.; Huang, R.; Mass, J.; Bisulca, C.; Bilderback, D.H.; Gruner, S.; Gao, N.

    2006-01-01

    A confocal X-ray fluorescence microscope was built at the Cornell High Energy Synchrotron Source (CHESS) to obtain compositional depth profiles of historic paintings. The microscope consists of a single-bounce, borosilicate monocapillary optic to focus the incident beam onto the painting and a commercial borosilicate polycapillary lens to collect the fluorescent X-rays. The resolution of the microscope was measured by scanning a variety of thin metal films through this confocal volume while monitoring the fluorescence signal. The capabilities of the technique were then probed using test paint microstructures with up to four distinct layers, each having a thickness in the range of 10-80 microns. Results from confocal XRF were compared with those from stand-alone XRF and visible light microscopy of the paint cross-sections. A large area, high-resolution scanner is currently being built to perform 3D scans on moderately sized paintings. (orig.)

  1. Ex vivo laser confocal microscopy findings of cultured Acanthamoeba trophozoites

    Directory of Open Access Journals (Sweden)

    Yamazaki N

    2012-08-01

    Full Text Available Natsuko Yamazaki,1 Akira Kobayashi,1 Hideaki Yokogawa,1 Yasuhisa Ishibashi,2 Yosaburo Oikawa,3 Masaharu Tokoro,4 Kazuhisa Sugiyama11Department of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan; 2Department of Ophthalmology, East Washinomiya Hospital, Kuki, Japan; 3Department of Medical Zoology, Kanazawa Medical University, Kahoku, Japan; 4Department of Parasitology, Kanazawa University Graduate School of Medical Science, Kanazawa, JapanPurpose: The purpose of the current study was to investigate ex vivo laser confocal microscopic findings of cultured Acanthamoeba trophozoites obtained from Acanthamoeba keratitis patients.Methods: Eight cultured samples of Acanthamoeba trophozoites from eight eyes of seven patients (mean age, 26.9 years; age range, 18–52 years were used. Seven samples were from corneal scrapings of Acanthamoeba keratitis patients and one sample was from the solution in a soft contact lens case. Ex vivo laser confocal microscopy was performed to qualitatively evaluate the shape and degree of light reflection of the living Acanthamoeba trophozoites.Results: Ex vivo laser confocal microscopy demonstrated highly reflective, high-contrast Acanthamoeba trophozoites with no walls (mean size, 25.4 µm; range, 17.1–58.5 µm. The shapes of the trophozoites were highly pleomorphic, and some showed characteristic acanthopodia by laser confocal microscopy.Conclusion: Ex vivo laser confocal microscopy was effective in demonstrating cultured Acanthamoeba trophozoites of various shapes and sizes. The observations of the current study may be helpful when similar structures are identified under in vivo conditions.Keywords: Acanthamoeba, trophozoite, laser confocal microscopy

  2. 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.

  3. 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.

  4. In vivo confocal microscopy for the detection of canine fungal keratitis and monitoring of therapeutic response.

    Science.gov (United States)

    Ledbetter, Eric C; Norman, Mary L; Starr, Jennifer K

    2016-05-01

    To describe in vivo corneal confocal microscopy of dogs during the clinical course of fungal keratitis and correlate findings with clinical evaluations and an ex vivo experimental canine fungal keratitis model. Seven dogs with naturally acquired fungal keratitis and ex vivo canine corneas experimentally infected with clinical fungal isolates. Dogs with naturally acquired fungal keratitis were examined by in vivo laser scanning confocal microscopy. Initial confocal microscopic examinations were performed to assist in establishing the diagnosis of fungal keratitis. Serial confocal microscopic examinations were performed to guide antifungal chemotherapy. Confocal microscopy images of canine corneal fungal isolates were obtained by examination of experimentally infected ex vivo canine corneas to corroborate in vivo findings. Fungi cultured and detected by PCR from canine corneal samples included Candida albicans, Fusarium incarnatum-equiseti, Malassezia pachydermatis, and a Rhodotorula sp. Linear, branching, interlocking, hyperreflective structures were detected by confocal microscopy in dogs with filamentous fungal keratitis and round to oval hyperreflective structures were detected in dogs with yeast fungal keratitis. Antifungal chemotherapy was associated with a progressive reduction in the distribution and density of corneal fungal elements, alterations to fungal morphology, decreased leukocyte numbers, restoration of epithelial layers, and an increased number of visible keratocyte nuclei. No dogs had a recurrence of fungal keratitis following medication discontinuation. Confocal microscopic fungal morphologies were similar between in vivo and ex vivo examinations. In vivo corneal confocal microscopy is a rapid method of diagnosing fungal keratitis in dogs and provides a noninvasive mechanism for monitoring therapeutic response. © 2015 American College of Veterinary Ophthalmologists.

  5. 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.

  6. EUS-Guided Needle-Based Confocal Laser Endomicroscopy

    DEFF Research Database (Denmark)

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

    2015-01-01

    the gut, providing further diagnostic and staging information. Confocal laser endomicroscopy (CLE) is a novel endoscopic method that enables imaging at a subcellular level of resolution during endoscopy, allowing up to 1000-fold magnification of tissue and providing an optical biopsy. A new procedure...... that has been developed in the past few years is needle-based confocal laser endomicroscopy (nCLE), which involves a mini-CLE probe that can be passed through a 1 9-gauge needle during EUS-FNA. This enables the real-time visualization of tissue at a microscopic level, with the potential to further improve....... The aim of this article is to review the role of EUS-guided nCLE in modern endoscopy and its implications in molecular imaging....

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

    Science.gov (United States)

    Chiu, L-D; Su, L; Reichelt, S; Amos, W B

    2012-05-01

    Shortly after its development, the white light supercontinuum laser was applied to confocal scanning microscopy as a more versatile substitute for the multiple monochromatic lasers normally used for the excitation of fluorescence. This light source is now available coupled to commercial confocal fluorescence microscopes. We have evaluated a supercontinuum laser as a source for a different purpose: confocal interferometric imaging of living cells and artificial models by interference reflection. We used light in the range 460-700 nm where this source provides a reasonably flat spectrum, and obtained images free from fringe artefacts caused by the longer coherence length of conventional lasers. We have also obtained images of cytoskeletal detail that is difficult to see with a monochromatic laser. © 2012 The Authors Journal of Microscopy © 2012 Royal Microscopical Society.

  8. Microscopic colitis

    DEFF Research Database (Denmark)

    Münch, A; Aust, D; Bohr, Jakob

    2012-01-01

    Microscopic colitis (MC) is an inflammatory bowel disease presenting with chronic, non-bloody watery diarrhoea and few or no endoscopic abnormalities. The histological examination reveals mainly two subtypes of MC, lymphocytic or collagenous colitis. Despite the fact that the incidence in MC has...... been rising over the last decades, research has been sparse and our knowledge about MC remains limited. Specialists in the field have initiated the European Microscopic Colitis Group (EMCG) with the primary goal to create awareness on MC. The EMCG is furthermore a forum with the intention to promote...

  9. Martian Microscope

    Science.gov (United States)

    2004-01-01

    The microscopic imager (circular device in center) is in clear view above the surface at Meridiani Planum, Mars, in this approximate true-color image taken by the panoramic camera on the Mars Exploration Rover Opportunity. The image was taken on the 9th sol of the rover's journey. The microscopic imager is located on the rover's instrument deployment device, or arm. The arrow is pointing to the lens of the instrument. Note the dust cover, which flips out to the left of the lens, is open. This approximated color image was created using the camera's violet and infrared filters as blue and red.

  10. Adaptive optics confocal microscopy using direct wavefront sensing.

    Science.gov (United States)

    Tao, Xiaodong; Fernandez, Bautista; Azucena, Oscar; Fu, Min; Garcia, Denise; Zuo, Yi; Chen, Diana C; Kubby, Joel

    2011-04-01

    Optical aberrations due to the inhomogeneous refractive index of tissue degrade the resolution and brightness of images in deep-tissue imaging. We introduce a confocal fluorescence microscope with adaptive optics, which can correct aberrations based on direct wavefront measurements using a Shack-Hartmann wavefront sensor with a fluorescent bead used as a point source reference beacon. The results show a 4.3× improvement in the Strehl ratio and a 240% improvement in the signal intensity for fixed mouse tissues at depths of up to 100 μm.

  11. Reflectance confocal microscopy: non-invasive distinction between actinic keratosis and squamous cell carcinoma

    NARCIS (Netherlands)

    Peppelman, M.; Nguyen, K.P.; Hoogedoorn, L.; Erp, P.E.J. van; Gerritsen, M.J.P.

    2015-01-01

    BACKGROUND: Early recognition of squamous cell carcinoma (SCC) is difficult. Non-invasive reflectance confocal microscopic (RCM) imaging of the skin is a promising diagnostic technique. Although several RCM features for SCC and AK have been described, it is not determined whether RCM has the ability

  12. The interference Linnik type microscope (IM) developed at Inmetro

    International Nuclear Information System (INIS)

    Malinovski, I; Couceiro, I B; Lima, M S; França, R S; Von der Weid, J P

    2015-01-01

    The interference Linnik type microscope was developed and it design was modernized to provide high resolution nanometrology facility. This microscope can perform length measurement directly relative to wavelength standards such as the frequency stabilized lasers. Thus, one can use interference microscope for primary calibration of the step heights. These step heights can be used to calibrate vertical axis of the other instruments like optical coherence tomography, atomic force microscope, confocal microscope, etc. In all cases the traceability to the National Institute of Metrology, Quality and Technology length primary standards was attempted to be established

  13. Shear Brillouin light scattering microscope.

    Science.gov (United States)

    Kim, Moonseok; Besner, Sebastien; Ramier, Antoine; Kwok, Sheldon J J; An, Jeesoo; Scarcelli, Giuliano; Yun, Seok Hyun

    2016-01-11

    Brillouin spectroscopy has been used to characterize shear acoustic phonons in materials. However, conventional instruments had slow acquisition times over 10 min per 1 mW of input optical power, and they required two objective lenses to form a 90° scattering geometry necessary for polarization coupling by shear phonons. Here, we demonstrate a confocal Brillouin microscope capable of detecting both shear and longitudinal phonons with improved speeds and with a single objective lens. Brillouin scattering spectra were measured from polycarbonate, fused quartz, and borosilicate in 1-10 s at an optical power level of 10 mW. The elastic constants, phonon mean free path and the ratio of the Pockels coefficients were determined at microscopic resolution.

  14. Submicron Confocal Raman Microscopy of Optical Holograms in Multicomponent Photopolymers

    Science.gov (United States)

    Kagan, C. R.; Harris, T. D.; Harris, A. L.; Schilling, M. L.

    1998-03-01

    We demonstrate submicron chemical imaging of optical holograms in multicomponent photopolymers using a scanning confocal Raman microscope. Our microscope is sensitive to the submicron, <1 percent concentration variations of the polymeric components that form the refractive index modulation responsible for hologram diffraction. Photopolymers are attractive media for holographic data storage, yet the mechanisms for generating the refractive index modulations responsible for hologram diffraction remain poorly understood. We obtain the first direct chemical evidence showing that these concentration modulations are established both by monomer diffusion and by polymer matrix swelling during hologram writing. Spatial variations in both density and composition contribute to the refractive index modulation. These measurements demonstrate the feasibility of submicron Raman microscopy in chemically imaging photodegradable organic and biological materials.

  15. Confocal Raman Microspectroscopy of Oral Streptococci

    Science.gov (United States)

    Beier, Brooke D.

    Raman spectroscopy has been used in a variety of applications throughout the field of biomedical optics. It has the ability to acquire chemically-specific information in a non-invasive manner, without the need for exogenous markers. This makes it useful in the identification of bacterial species, as well as in the study of tissues and other cells. In this work, a species identification model has been created in order to discriminate between the oral bacterial species Streptococcus sanguinis and Streptococcus mutans. These are two of the most prevalent species within the human mouth and their relative concentrations can be an indicator of a patient's oral health and risk of tooth decay. They are predominantly found within plaque on the tooth's surface. To study a simplified model for dental plaque, we have examined S. sanguinis and S. mutans grown in biofilm forms. Raman spectroscopy has been implemented here through a confocal microscope. The optical system has been equipped with computationally controlled stages to allow for automated scanning, including autofocusing to probe a consistent depth within a sample. A spectrum has been acquired from each position within a scan and sent for spectral preprocessing before being submitted for species identification. This preprocessing includes an algorithm that has been developed to remove fluorescence features from known contaminants within the confocal volume, to include signal from a fluorescent substrate. Species classification has been accomplished using a principal component score-fed logistic regression model constructed from a variety of biofilm samples that have been transferred and allowed to dry, as might occur with the study of plaque samples. This binary classification model has been validated on other samples with identical preparations. The model has also been transferred to determine the species of hydrated biofilms studied in situ. Artificially mixed biofilms have been examined to test the spatial

  16. Specificity of in vivo confocal cornea microscopy in Acanthamoeba keratitis.

    Science.gov (United States)

    Füst, Ágnes; Tóth, Jeannette; Simon, Gyula; Imre, László; Nagy, Zoltán Z

    2017-01-19

    To report on the presence of 4 different structures visualized by confocal microscopy in patients whose clinical presentation suggested infection by Acanthamoeba. Data and charts of 28 consecutive patients were analyzed in a retrospective study. Four types of structures were recognized by confocal microscopy performed with HRT II Rostock Cornea Module: trophozoites, double-walled cysts, signet rings, and bright spots. The 28 patients (mean age 30.8 years, range 17-61 years, 10 male, 18 female) were divided into 4 groups according to the diagnosis established later by microscopic examination of smear, culture, response to therapy, and the course of keratitis. The 4 groups were Acanthamoeba keratitis (AK), Acanthamoeba suspect (AK-suspect), bacterial keratitis (BK), and fungal keratitis (FK). The rate of patients in AK, AK-suspect, FK, and BK groups where bright spots were found were 100%, 100%, 40%, and 55%, respectively. The sensitivity of presence of bright spots in the in vivo confocal microscopy in Acanthamoeba keratitis was 100% (95% confidence interval [CI] 73.5% to 100.00%) and specificity was 50% (CI 24.7% to 75.4%). When cases where the only signs of Acanthamoeba were bright spots were excluded, and only those cases were counted where any of cysts, trophozoites, or signet rings were also found, the sensitivity was 67% (95% CI 34. 9% to 90.1%) and the specificity was 94% (95% CI 69.8% to 99.8%). The relatively high rate of bright spots in non-Acanthamoeba keratitis challenges the assumption that bright spots seen by confocal microscopy are a specific indication of Acanthamoeba keratitis.

  17. QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY

    Directory of Open Access Journals (Sweden)

    Merete Krog Raarup

    2011-05-01

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

  18. 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

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

    Science.gov (United States)

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

    2013-09-01

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

  20. 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.

  1. Confocal microscopy findings of Acanthamoeba keratitis.

    Science.gov (United States)

    Pfister, D R; Cameron, J D; Krachmer, J H; Holland, E J

    1996-02-01

    Tandem scanning confocal microscopy was performed on two patients with Acanthamoeba keratitis to provide images detailing characteristic findings of the disease. Although tandem scanning confocal microscopy of Acanthamoeba has been described in previous reports, Acanthamoeba keratitis has not been fully characterized with this instrument. In vivo confocal micrographs showed the double-walled structure of the Acanthamoeba cyst and associated radial keratoneuritis (perineuritis). We reviewed the records of two patients with a clinical diagnosis of Acanthamoeba keratitis, one with culture-proven Acanthamoeba and the other with a suspected Acanthamoeba infection. Slit-lamp biomicroscopy and in vivo tandem scanning confocal microscopy were performed. The images obtained were compared with images from patients without corneal disease. High-contrast round bodies suggestive of Acanthamoeba cysts, as previously described, and irregular forms suggestive of Acanthamoeba trophozoites were found by tandem scanning confocal microscopy. Additionally, we showed conclusively that under certain circumstances (that is, corneal scarring) tandem scanning confocal microscopy can resolve the double-walled structure of the Acanthamoeba ectocyst surrounding the endocyst. Furthermore, radial keratoneuritis was demonstrated, consisting of an irregularly swollen nerve fiber with probable amoebic infiltration. Confocal microscopy can be a useful, noninvasive imaging technique helpful in the study, diagnosis, and treatment of Acanthamoeba keratitis.

  2. Line-scanning tomographic optical microscope with isotropic transfer function

    International Nuclear Information System (INIS)

    Gajdátsy, Gábor; Dudás, László; Erdélyi, Miklós; Szabó, Gábor

    2010-01-01

    An imaging method and optical system, referred to as a line-scanning tomographic optical microscope (LSTOM) using a combination of line-scanning technique and CT reconstruction principle, is proposed and studied theoretically and experimentally. In our implementation a narrow focus line is scanned over the sample and the reflected light is measured in a confocal arrangement. One such scan is equivalent to a transverse projection in tomography. Repeating the scanning procedure in several directions, a number of transverse projections are recorded from which the image can be obtained using conventional CT reconstruction algorithms. The resolution of the image is independent of the spatial dimensions and structure of the applied detector; furthermore, the transfer function of the system is isotropic. The imaging performance of the implemented confocal LSTOM was compared with a point-scanning confocal microscope, based on recorded images. These images demonstrate that the resolution of the confocal LSTOM exceeds (by 15%) the resolution limit of a point-scanning confocal microscope

  3. 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.

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

    Science.gov (United States)

    Yang, Yong-fa; Li, Qi

    2014-12-01

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

  5. 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.

  6. Confocal Laser Endomicroscopy in Inflammatory Bowel Disease

    DEFF Research Database (Denmark)

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

    2015-01-01

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

  7. In-vivo immunofluorescence confocal microscopy of herpes simplex virus type 1 keratitis

    Science.gov (United States)

    Kaufman, Stephen C.; Laird, Jeffery A.; Beuerman, Roger W.

    1996-05-01

    The white-light confocal microscope offers an in vivo, cellular-level resolution view of the cornea. This instrument has proven to be a valuable research and diagnostic tool for the study of infectious keratitis. In this study, we investigate the direct visualization of herpes simplex virus type 1 (HSV-1)-infected corneal epithelium, with in vivo confocal microscopy, using HSV-1 immunofluorescent antibodies. New Zealand white rabbits were infected with McKrae strain of HSV-1 in one eye; the other eye of each rabbit was used as an uninfected control. Four days later, the rabbits were anesthetized and a cellulose sponge was applied to each cornea, and a drop of direct HSV fluorescein-tagged antibody was placed on each sponge every 3 to 5 minutes for 1 hour. Fluorescence confocal microscopy was then performed. The HSV-infected corneas showed broad regions of hyperfluorescent epithelial cells. The uninfected corneas revealed no background fluorescence. Thus, using the confocal microscope with a fluorescent cube, we were able to visualize HSV-infected corneal epithelial cells tagged with a direct fluorescent antibody. This process may prove to be a useful clinical tool for the in vivo diagnosis of HSV keratitis.

  8. Scanner component and head development for confocal microscopy using moving mirror technology

    Science.gov (United States)

    Loney, Gregory C.

    1993-12-01

    One of the challenges in designing a confocal microscope is choosing the scan system configuration. The selection is based largely on the microscope application and involves a few distinct schemes. One scheme, moving mirror using galvanometer and resonant scanners, has been shown to offer an excellent solution exhibited by the large number of commercial systems which utilize them. Perceived shortcomings, such as slow image acquisition, are being dispelled due to the advent of large angle, high frequency resonant scanners. These newer devices offer near video rate performance at good scan efficiency.

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    Kobayashi A

    2014-02-01

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

  11. Basic principles of confocal microscopy of cаlcium signals

    Directory of Open Access Journals (Sweden)

    В. М. Шкрыль

    2017-12-01

    Full Text Available Background: The study of fast processes occurring in living cells, for example, the dynamics of calcium ions or other ions, is actual for modern biophysics, physiology and medicine and can be carried out by laser scanning confocal microscopy. Confocal microscopy as a method of studying biological objects has a number of features requiring an understanding of the delicate mechanisms underlying it. Objectives: The aim of the work is the description of principles and details of measurements of calcium signals in the living cells using laser scanning confocal microscopy and fluorescent indicators. Materials and methods: The analysis of the methodological and practical aspects of studies of calcium signals was carried out by confocal microscopy in the work. Results: Confocal microscopy allows to study the changes in the concentration of free calcium within the cell and even a small part of it by a fluorescent dye, which increases the contrast in comparison with conventional fluorescence microscopy through an additional confocal aperture located in the front of the detector and the use of laser light source and scanning the object. To register calcium signals it is necessary to make a selection of a number of adequate parameters: the registration frequency, the size of a single scanning element (pixel, the sensitivity of the detector, the intensity of laser irradiation, the diameter of the confocal gap, the corresponding filters for exciting and emitting the fluorescent dye used and the corresponding dichroic mirror. An important stage of a setup of a confocal system is the determination of the value of the scattering function of a point. Compensation of the process of bleaching of the fluorescence dye and reduction of phototoxicity, minimization of the scattering process of the image allow to increase a reproducibility of the experiments. Conclusions: Using modern laser scanning confocal microscopes for registration of calcium signals from a small

  12. Microscope spectrometer for light scattering investigations.

    Science.gov (United States)

    Barbara, Aude; Lopez-Rios, Tomas; Dumont, Sylvain; Gay, Frédéric; Quémerais, Pascal

    2010-08-01

    We describe a setup including a microscope to study volumes of a few mum(3) by static and dynamic light scattering (DLS) in a backscattering configuration. Light scattered by individual objects of micrometric size can be analyzed in the 400-800?nm spectral range. This setup can also be employed to study both diluted and concentrated colloidal solutions by DLS measurements. For diluted solutions we found evidence of the fluctuations of the number of particles in a confocal volume. We discuss their contribution to the autocorrelation function of the scattered intensity measured as a function of time.

  13. Confocal pattern period in multiple-aperture confocal imaging systems with coherent illumination.

    Science.gov (United States)

    Liang, M; Stehr, R L; Krause, A W

    1997-06-01

    We have built a programmable multiple-aperture confocal imaging system that uses a spatial light modulator (a Texas Instruments digital micromirror device). Excellent axial resolution and confocal imaging quality have been experimentally demonstrated with this system, even when the distance between adjacent ON pixels is four times the size of the pixel aperture. By contrast, typical pinhole-based systems (e.g., a Nipkow disk) employ an adjacent aperture distance-to-size ratio of 10x . We have achieved improvement over such systems by 6.25x in both light-utilization efficiency and confocal image rate without adding more components and extra processing time.

  14. Coherent artifact suppression in line-field reflection confocal microscopy using a low spatial coherence light source.

    Science.gov (United States)

    Liu, Changgeng; Cao, Hui; Choma, Michael A

    2016-10-15

    Line-field reflection confocal microscopy (LF-RCM) has the potential to add a dimension of parallelization to traditional confocal microscopy while reducing the need for two-axis beam scanning. LF-RCM systems often employ light sources with a high degree of spatial coherence. This high degree of spatial coherence potentially leads to unwanted coherent artifact in the setting of nontrivial sample scattering. Here, we (a) confirm that a coherent artifact is a nontrivial problem in LF-RCM when using spatially coherent light, and (b) demonstrate that such a coherent artifact can be mitigated through the use of reduced spatial coherence line-field sources. We demonstrate coherent noise suppression in a full-pupil line-field confocal microscope using a large number of mutually incoherent emitters from a vertical-cavity surface-emitting lasers (VCSEL) array. The coherent noise from a highly scattering sample is significantly suppressed by the use of this synthesized reduced spatial coherence light source compared to a fully coherent light source. Lastly, with scattering samples, the axial confocality of line-field confocal microscopy is compromised independent of the source spatial coherence, as demonstrated by our experimental result. Our results highlight the importance of spatial coherence engineering in parallelized reflection confocal microscopy.

  15. Fast imaging with inelastically scattered electrons by off-axis chromatic confocal electron microscopy.

    Science.gov (United States)

    Zheng, Changlin; Zhu, Ye; Lazar, Sorin; Etheridge, Joanne

    2014-04-25

    We introduce off-axis chromatic scanning confocal electron microscopy, a technique for fast mapping of inelastically scattered electrons in a scanning transmission electron microscope without a spectrometer. The off-axis confocal mode enables the inelastically scattered electrons to be chromatically dispersed both parallel and perpendicular to the optic axis. This enables electrons with different energy losses to be separated and detected in the image plane, enabling efficient energy filtering in a confocal mode with an integrating detector. We describe the experimental configuration and demonstrate the method with nanoscale core-loss chemical mapping of silver (M4,5) in an aluminium-silver alloy and atomic scale imaging of the low intensity core-loss La (M4,5@840  eV) signal in LaB6. Scan rates up to 2 orders of magnitude faster than conventional methods were used, enabling a corresponding reduction in radiation dose and increase in the field of view. If coupled with the enhanced depth and lateral resolution of the incoherent confocal configuration, this offers an approach for nanoscale three-dimensional chemical mapping.

  16. Application of a novel confocal imaging technique for early the detection of dental decay

    Science.gov (United States)

    Rousseau, Christel; Girkin, John M.; Vaidya, Shilpa; Hall, Andrew F.; Whitters, C. J.; Creanor, Steve L.

    2002-06-01

    In order to stop or prevent the progression of dental disease, early detection and quantification of decay are crucially important. Dental decay (caries) detection methods have traditionally involved clinical examination by eye, using probes and dental radiography, but up to 60% of lesions are missed. What the dentist requires is a cheap, reliable method of detection of early disease, ideally with information on the depth and rate of growth or healing. Conventional commercial scanning confocal microscopes are unsuitable for use on dental patients. We report on a fibre optic based confocal microscope designed for in vivo examination of caries lesions. The system utilizes a common fibre both as the source and to detect the reflected confocal signal. The initial system has been optimized using dielectric mirrors and the thickness of the stack has been measured with high precision. Dental samples have been examined and the system has been demonstrated to provide information on the depth and mineral loss of a lesion. Fibre optic microscopy (FOCM) demonstrates a practical route to developing an in vivo caries profiler. In this paper, the FOCM and its applications in caries detection are described and the potential of this scheme as a practical dental probe is discussed.

  17. 3D imaging of neutron tracks using confocal microscopy

    Science.gov (United States)

    Gillmore, Gavin; Wertheim, David; Flowers, Alan

    2016-04-01

    Neutron detection and neutron flux assessment are important aspects in monitoring nuclear energy production. Neutron flux measurements can also provide information on potential biological damage from exposure. In addition to the applications for neutron measurement in nuclear energy, neutron detection has been proposed as a method of enhancing neutrino detectors and cosmic ray flux has also been assessed using ground-level neutron detectors. Solid State Nuclear Track Detectors (or SSNTDs) have been used extensively to examine cosmic rays, long-lived radioactive elements, radon concentrations in buildings and the age of geological samples. Passive SSNTDs consisting of a CR-39 plastic are commonly used to measure radon because they respond to incident charged particles such as alpha particles from radon gas in air. They have a large dynamic range and a linear flux response. We have previously applied confocal microscopy to obtain 3D images of alpha particle tracks in SSNTDs from radon track monitoring (1). As a charged particle traverses through the polymer it creates an ionisation trail along its path. The trail or track is normally enhanced by chemical etching to better expose radiation damage, as the damaged area is more sensitive to the etchant than the bulk material. Particle tracks in CR-39 are usually assessed using 2D optical microscopy. In this study 6 detectors were examined using an Olympus OLS4100 LEXT 3D laser scanning confocal microscope (Olympus Corporation, Japan). The detectors had been etched for 2 hours 50 minutes at 85 °C in 6.25M NaOH. Post etch the plastics had been treated with a 10 minute immersion in a 2% acetic acid stop bath, followed by rinsing in deionised water. The detectors examined had been irradiated with a 2mSv neutron dose from an Am(Be) neutron source (producing roughly 20 tracks per mm2). We were able to successfully acquire 3D images of neutron tracks in the detectors studied. The range of track diameter observed was between 4

  18. Confocal Microscopy for Real Time Detection of Oral Cavity Neoplasia

    National Research Council Canada - National Science Library

    Clark, Anne

    2003-01-01

    .... Reflectance confocal images were compared to histologic images from the same sample to determine which tissue features contribute to image contrast and can be potentially imaged using in vivo confocal microscopy...

  19. Transmission positron microscopes

    International Nuclear Information System (INIS)

    Doyama, Masao; Kogure, Yoshiaki; Inoue, Miyoshi; Kurihara, Toshikazu; Yoshiie, Toshimasa; Oshima, Ryuichiro; Matsuya, Miyuki

    2006-01-01

    Immediate and near-future plans for transmission positron microscopes being built at KEK, Tsukuba, Japan, are described. The characteristic feature of this project is remolding a commercial electron microscope to a positron microscope. A point source of electrons kept at a negative high voltage is changed to a point source of positrons kept at a high positive voltage. Positional resolution of transmission microscopes should be theoretically the same as electron microscopes. Positron microscopes utilizing trapping of positrons have always positional ambiguity due to the diffusion of positrons

  20. 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.

  1. Compensation of phase aberration by using a virtual confocal scheme in digital holographic microscopy.

    Science.gov (United States)

    Chew, Yang-Kun; Shiu, Min-Tzung; Wang, Je-Chung; Chang, Chi-Ching

    2014-09-20

    This work presents cost-effective, simple arbitrary phase-step digital holographic microscopy to suppress both zero-order and twin-image terms. A virtual confocal offset lens under in-line configuration is also used to compensate for the introduced quadratic phase by using a microscope objective lens. In addition to reducing the difficulties of physical confocal configurations, the proposed method significantly increases the magnification power, ultimately achieving the purposes of an optical zoom. An attempt is also made to reduce the noise interference of a high magnification system by developing a long focal lens to reduce light detection size, subsequently gaining an approximately plane wave light source to illuminate the object within the effective depth of focus. Experimental results indicate that the proposed high magnification system can be elevated with low noise interference, and image reconstruction without quadratic phase terms.

  2. Fluorescence lifetime measurement with confocal endomicroscopy for direct analysis of tissue biochemistry in vivo

    Directory of Open Access Journals (Sweden)

    Youngjae Won

    2016-08-01

    Full Text Available Confocal endomicroscopy is a powerful tool for in vivo real-time imaging at cellular resolution inside a living body without tissue resection. Microscopic fluorescence lifetime measurement can provide information about localized biochemical conditions such as pH and the concentrations of oxygen and calcium. We hypothesized that combining these techniques could assist accurate cancer discrimination by providing both biochemical and morphological information. We designed a dual-mode experimental setup for confocal endomicroscopic imaging and fluorescence lifetime measurement and applied it to a mouse xenograft model of activated human pancreatic cancer generated by subcutaneous injection of AsPC-1 tumor cells. Using this method with pH-sensitive sodium fluorescein injection, we demonstrated discrimination between normal and cancerous tissues in a living mouse. With further development, this method may be useful for clinical cancer detection.

  3. 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...

  4. Analysis of multi-dimensional confocal images

    International Nuclear Information System (INIS)

    Samarabandu, J.K.; Acharya, R.; Edirisinghe, C.D.; Cheng, P.C.; Lin, T.H.

    1991-01-01

    In this paper, a confocal image understanding system is developed which used the blackboard model of problem solving to achieve computerized identification and characterization of confocal fluorescent images (serial optical sections). The system is capable of identifying a large percentage of structures (e.g. cell nucleus) in the presence of background noise and non specific staining of cellular structures. The blackboard architecture provides a convenient framework within which a combination of image processing techniques can be applied to successively refine the input image. The system is organized to find the surfaces of highly visible structures first, using simple image processing techniques and then to adjust and fill in the missing areas of these object surfaces using external knowledge, and a number of more complex image processing techniques when necessary. As a result, the image analysis system is capable of obtaining morphometrical parameters such as surface area, volume and position of structures of interest automatically

  5. 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.

  6. In vivo visualization of honeycomb pattern, cobblestone pattern, ringed pattern, and dermal papillae by confocal laser scanning microscopy.

    Science.gov (United States)

    Xiang, W; Peng, J; Song, X; Xu, A; Zhang, D; Liu, J; Bi, Z

    2016-02-01

    In vivo confocal laser scanning microscopy (CLSM) represents a promising technique for noninvasive visualization of lesions. Honeycomb pattern, cobblestone pattern, ringed pattern and dermal papillae are terms commonly used to describe CLSM architectures. Understanding of these four terms individually or in combination on confocal images facilitates the diagnosis and differential diagnosis of skin diseases. Seventy-eight patients and 5 healthy controls were enrolled in the study. Their lesions were imaged with the VivaScope 1500 reflectance confocal microscope. Concerning these four terms mentioned above, they had different features in different skin diseases and played an important role in the understanding of skin diseases individually or in combination. By understanding the CLSM features of honeycomb pattern, cobblestone pattern, ringed pattern and dermal papillae individually or in combination, the findings support the roles of these characteristic architectures in diagnosis and differential diagnosis of skin diseases. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. The Scanning Optical Microscope.

    Science.gov (United States)

    Sheppard, C. J. R.

    1978-01-01

    Describes the principle of the scanning optical microscope and explains its advantages over the conventional microscope in the improvement of resolution and contrast, as well as the possibility of producing a picture from optical harmonies generated within the specimen.

  8. 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.

  9. Confocal laser scanning microscopy in study of bone calcification

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-12-01

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

  10. Wettability study and surface characterizacion by confocal laser scanning microscopy of rotary-peeled wood veneers

    OpenAIRE

    Vázquez Uña, Gonzalo; Galiñanes Bello, Cristina; Freire Leira, María Sonia; Antorrena Álvarez, Gervasio; González Álvarez, Julia

    2011-01-01

    [ES]El objetivo de este trabajo fue estudiar las propiedades superficiales de chapas obtenidas por desenrollo de cinco especies de maderas duras: okume, ayous, chopo, fromager y eucalipto, y comparar el comportamiento de ambas caras de las chapas. Se han determinado los ángulos de contacto de equilibrio y se ha determinado la energía libre superficial de la madera así como sus componentes polar y dispersa. Asimismo, mediante microscopía de barrido láser confocal se ha evaluado la rugosidad su...

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Microscopia confocal en córneas de cien ojos sanos Confocal microscopy results of one hundred healthy eye corneas

    Directory of Open Access Journals (Sweden)

    Zulema Gómez Castillo

    2012-06-01

    disease notified from May 2007 to May 2008 were considered. Results: The 100 studied eyes came from apparently healthy patients, and the pachymetry values of 64 of them were above the average. Since they were healthy corneas, the 3 types of epithelial cells as well as the keratocytes were present in almost all the patients and at different depths of the corneal stroma, respectively. Most of the studied eyes had endothelial cell counts above 2 500, a figure within the normal range, and there were nerve fibers in each of their layers. Conclusion: Confocal microscope is a new tool that allows in vivo observation of corneal histology and thus supplements conventional biomicroscopy observations. It contributes to better understanding of corneal pathology, with a view to prophylactically and therapeutically acting upon corneal pathologies during their follow-up and evolution.

  13. Confocal profilometer with nanometric vertical resolution

    International Nuclear Information System (INIS)

    Butler, D.J.; Horsfall, A.; Hrynevych, M.; Kearney, P.D.; Nugent, K.A.

    1992-01-01

    An optical system is described which is based on the method of Kobayashi et al. The instrument is capable of simultaneous confocal imaging and profilometry with nanometric (nm) vertical resolution. The profile is independent of the reflectivity of the sample and is obtained by raster-scanning the sample without active feedback. It is shown that the vertical resolution of this technique may be extended down to 1 nm and have recorded images with a resolution of better than 3 nm. The concept and design is considerably simpler than most optical profilometers and may represent the most affordable method of profilometry with nanometric resolution available. 15 refs., 6 figs

  14. Cryogenic immersion microscope

    Science.gov (United States)

    Le Gros, Mark; Larabell, Carolyn A.

    2010-12-14

    A cryogenic immersion microscope whose objective lens is at least partially in contact with a liquid reservoir of a cryogenic liquid, in which reservoir a sample of interest is immersed is disclosed. When the cryogenic liquid has an index of refraction that reduces refraction at interfaces between the lens and the sample, overall resolution and image quality are improved. A combination of an immersion microscope and x-ray microscope, suitable for imaging at cryogenic temperatures is also disclosed.

  15. Objectives comparison in a confocal microscope using pseuso-random roughness artefacts

    DEFF Research Database (Denmark)

    Quagliotti, Danilo; Tosello, Guido; Hansen, Hans Nørgaard

    The increasing importance of optical microscopy in the geometrical and dimensional assessment of structured and freeform surfaces has easily overcome many difficulties related to the use of contact instruments. Nonetheless, some concerns related to optical instruments when measuring such surfaces...

  16. Microscopía confocal en las enfermedades ampollosas subepidérmicas autoinmunes

    OpenAIRE

    Campos Domínguez, Minia

    2016-01-01

    Las enfermedades ampollosas subepidérmicas autoinmunes constituyen un complejo grupo de trastornos cutáneos en cuya patogenia se rompe la unión dermoepidérmica. Su alta morbilidad se debe a la afectación de grandes áreas cutáneas por ampollas, erosiones o cicatrices y a la posible afectación de las mucosas oral, conjuntival, laríngea, esofágica y genitourinaria. Además, se ha demostrado su asociación con enfermedades neurológicas degenerativas y neoplasias. Existe un gran solapamiento clínico...

  17. 3D Segmentations of Neuronal Nuclei from Confocal Microscope Image Stacks

    Directory of Open Access Journals (Sweden)

    Antonio eLaTorre

    2013-12-01

    Full Text Available In this paper, we present an algorithm to create 3D segmentations of neuronal cells from stacks of previously segmented 2D images. The idea behind this proposal is to provide a general method to reconstruct 3D structures from 2D stacks, regardless of how these 2D stacks have been obtained. The algorithm not only reuses the information obtained in the 2D segmentation, but also attempts to correct some typical mistakes made by the 2D segmentation algorithms (for example, under segmentation of tightly-coupled clusters of cells. We have tested our algorithm in a real scenario --- the segmentation of the neuronal nuclei in different layers of the rat cerebral cortex. Several representative images from different layers of the cerebral cortex have been considered and several 2D segmentation algorithms have been compared. Furthermore, the algorithm has also been compared with the traditional 3D Watershed algorithm and the results obtained here show better performance in terms of correctly identified neuronal nuclei.

  18. Analytical Electron Microscope

    Data.gov (United States)

    Federal Laboratory Consortium — The Titan 80-300 is a transmission electron microscope (TEM) equipped with spectroscopic detectors to allow chemical, elemental, and other analytical measurements to...

  19. Compressive confocal microscopy: 3D reconstruction algorithms

    Science.gov (United States)

    Ye, P.; Paredes, J. L.; Wu, Y.; Chen, C.; Arce, G. R.; Prather, D. W.

    2009-02-01

    In this paper, a new approach for Confocal Microscopy (CM) based on the framework of compressive sensing is developed. In the proposed approach, a point illumination and a random set of pinholes are used to eliminate out-of-focus information at the detector. Furthermore, a Digital Micromirror Device (DMD) is used to efficiently scan the 2D or 3D specimen but, unlike the conventional CM that uses CCD detectors, the measured data in the proposed compressive confocal microscopy (CCM) emerge from random sets of pinhole illuminated pixels in the specimen that are linearly combined (projected) and measured by a single photon detector. Compared to conventional CM or programmable array microscopy (PAM), the number of measurements needed for nearly perfect reconstruction in CCM is significantly reduced. Our experimental results are based on a testbed that uses a Texas Instruments DMD (an array of 1024×768 13.68×13.68 μm2 mirrors) for computing the linear projections of illuminated pixels and a single photon detector is used to obtain the compressive sensing measurement. The position of each element in the DMD is defined by the compressed sensing measurement matrices. Threedimensional image reconstruction algorithms are developed that exploit the inter-slice spatial image correlation as well as the correlation between different 2D slices. A comprehensive performance comparison between several binary projection patterns is shown. Experimental and simulation results are provided to illustrate the features of the proposed systems.

  20. Refractive index measurement based on confocal method

    Science.gov (United States)

    An, Zhe; Xu, XiPing; Yang, JinHua; Qiao, Yang; Liu, Yang

    2017-10-01

    The development of transparent materials is closed to optoelectronic technology. It plays an increasingly important role in various fields. It is not only widely used in optical lens, optical element, optical fiber grating, optoelectronics, but also widely used in the building material, pharmaceutical industry with vessel, aircraft windshield and daily wear glasses.Regard of solving the problem of refractive index measurement in optical transparent materials. We proposed that using the polychromatic confocal method to measuring the refractive index of transparent materials. In this article, we describes the principle of polychromatic confocal method for measuring the refractive index of glass,and sketched the optical system and its optimization. Then we establish the measurement model of the refractive index, and set up the experimental system. In this way, the refractive index of the glass has been calibrated for refractive index experiment. Due to the error in the experimental process, we manipulated the experiment data to compensate the refractive index measurement formula. The experiment taking the quartz glass for instance. The measurement accuracy of the refractive index of the glass is +/-1.8×10-5. This method is more practical and accurate, especially suitable for non-contact measurement occasions, which environmental requirements is not high. Environmental requirements are not high, the ordinary glass production line up to the ambient temperature can be fully adapted. There is no need for the color of the measured object that you can measure the white and a variety of colored glass.

  1. Fungal keratitis - improving diagnostics by confocal microscopy.

    Science.gov (United States)

    Nielsen, E; Heegaard, S; Prause, J U; Ivarsen, A; Mortensen, K L; Hjortdal, J

    2013-09-01

    Introducing a simple image grading system to support the interpretation of in vivo confocal microscopy (IVCM) images in filamentous fungal keratitis. 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. 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. 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 12-69), 6 out of 17 (35%) cultures were positive and a total of 6/7 (86%) IVCM scans were positive. Three different categories of IVCM results for the grading of diagnostic certainty were formed. IVCM is a valuable tool for diagnosing filamentous fungal keratitis. In order to improve the reliability of IVCM, we suggest implementing a simple and clinically applicable grading system for aiding the interpretation of IVCM images of filamentous fungal keratitis.

  2. Fungal Keratitis - Improving Diagnostics by Confocal Microscopy

    Directory of Open Access Journals (Sweden)

    Esben Nielsen

    2013-12-01

    Full Text Available 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 12-69, 6 out of 17 (35% cultures were positive and a total of 6/7 (86% IVCM scans were positive. Three different categories of IVCM results for the grading of diagnostic certainty were formed. Conclusion: IVCM is a valuable tool for diagnosing filamentous fungal keratitis. In order to improve the reliability of IVCM, we suggest implementing a simple and clinically applicable grading system for aiding the interpretation of IVCM images of filamentous fungal keratitis.

  3. 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.

  4. Transverse vibration of spinning disk with attached distributed patch ...

    African Journals Online (AJOL)

    Free and forced transverse vibration characteristics of a thin spinning disc attached to a rigid core have been investigated by finite element analysis using ANSYS software. The effect of discrete point masses and patches of distributed masses attached at the periphery of the plate on free and forced vibration behavior of a ...

  5. Minimizing light exposure with the programmable array microscope.

    Science.gov (United States)

    Caarls, W; Rieger, B; De Vries, A H B; Arndt-Jovin, D J; Jovin, T M

    2011-01-01

    The exposure of fluorophores to intense illumination in a microscope often results in photobleaching and phototoxicity, thus constituting a major limiting factor in time lapse live cell or single molecule imaging. Laser scanning confocal microscopes are particularly prone to this problem, inasmuch as they require high irradiances to compensate for the inherently low duty cycle of point scanning systems. In the attempt to maintain adequate speed and signal-to-noise ratios, the fluorophores are often driven into saturation, thereby generating a nonlinear response. One approach for reducing photodegradation in the laser scanning confocal microscope is represented by controlled light exposure microscopy, introduced by Manders and colleagues. The strategy is to reduce the illumination intensity in both background areas (devoid of information) as well as in bright foreground regions, for which an adequate signal-to-noise ratio can be achieved with lower excitation levels than those required for the less intense foreground pixels/voxels. Such a variable illumination scheme can also be exploited in widefield microscopes that employ lower irradiance but higher illumination duty cycles. We report here on the adaptation of the controlled light exposure microscopy principle to the programmable array microscope, which achieves optical sectioning by use of a spatial light modulator (SLM) in an image plane as a programmable mask for illumination and conjugate (and nonconjugate) detection. By incorporating the basic controlled light exposure microscopy concept for minimizing exposure, we have obtained a reduction in the rate of photobleaching of up to ~5-fold, while maintaining an image quality comparable to regular imaging with the programmable array microscope. © 2010 The Authors Journal of Microscopy © 2010 The Royal Microscopical Society.

  6. Optical microscopic imaging based on VRML language

    Science.gov (United States)

    Zhang, Xuedian; Zhang, Zhenyi; Sun, Jun

    2009-11-01

    As so-called VRML (Virtual Reality Modeling Language), is a kind of language used to establish a model of the real world or a colorful world made by people. As in international standard, VRML is the main kind of program language based on the "www" net building, which is defined by ISO, the kind of MIME is x-world or x-VRML. The most important is that it has no relationship with the operating system. Otherwise, because of the birth of VRML 2.0, its ability of describing the dynamic condition gets better, and the interaction of the internet evolved too. The use of VRML will bring a revolutionary change of confocal microscope. For example, we could send different kinds of swatch in virtual 3D style to the net. On the other hand, scientists in different countries could use the same microscope in the same time to watch the same samples by the internet. The mode of sending original data in the model of text has many advantages, such as: the faster transporting, the fewer data, the more convenient updating and fewer errors. In the following words we shall discuss the basic elements of using VRML in the field of Optical Microscopic imaging.

  7. Poly(diacetylene) Monolayers Studied with a Fluorescence Scanning Near-Field Optical Microscope

    NARCIS (Netherlands)

    Moers, Marco H.P.; Moers, M.H.P.; Gaub, Hermann E.; van Hulst, N.F.

    1994-01-01

    A novel and powerful method to study the optical properties of thin lipid films which a resolution superior to confocal microscopy is presented. With a scanning near-field optical microscope, fluorescence images of a Langmuir-Blodgett film of diethylene glycol diamine pentacosadiynoic amide are

  8. The Light Microscope.

    Science.gov (United States)

    Baker, W. L.

    1995-01-01

    Describes the function of the various parts of the microscope and their integration in the formation of an optical image. Presents a procedure for setting up a microscope to obtain maximum resolution and contrast for each objective lens at all magnifications. (JRH)

  9. Mailing microscope slides

    Science.gov (United States)

    Many insects feed agriculturally important crops, trees, and ornamental plants and cause millions of dollars of damage annually. Identification for some of these require the preparation of a microscope slide for examination. There are times when a microscope slide may need to be sent away to a speci...

  10. Quantification of whey in fluid milk using confocal Raman microscopy and artificial neural network.

    Science.gov (United States)

    Alves da Rocha, Roney; Paiva, Igor Moura; Anjos, Virgílio; Furtado, Marco Antônio Moreira; Bell, Maria José Valenzuela

    2015-06-01

    In this work, we assessed the use of confocal Raman microscopy and artificial neural network as a practical method to assess and quantify adulteration of fluid milk by addition of whey. Milk samples with added whey (from 0 to 100%) were prepared, simulating different levels of fraudulent adulteration. All analyses were carried out by direct inspection at the light microscope after depositing drops from each sample on a microscope slide and drying them at room temperature. No pre- or posttreatment (e.g., sample preparation or spectral correction) was required in the analyses. Quantitative determination of adulteration was performed through a feed-forward artificial neural network (ANN). Different ANN configurations were evaluated based on their coefficient of determination (R2) and root mean square error values, which were criteria for selecting the best predictor model. In the selected model, we observed that data from both training and validation subsets presented R2>99.99%, indicating that the combination of confocal Raman microscopy and ANN is a rapid, simple, and efficient method to quantify milk adulteration by whey. Because sample preparation and postprocessing of spectra were not required, the method has potential applications in health surveillance and food quality monitoring. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  11. Effect of hygroscopic expansion of resin filling on interfacial gap and sealing: a confocal microscopy study.

    Science.gov (United States)

    Rosales-Leal, Juan I; Castillo-Salmerón, Ramón Del; Molino-Serrano, María A; González-Moreira, Humberto; Cabrerizo-Vílchez, Miguel A

    2013-10-01

    To measure dimensional changes due to hygroscopic expansion and their effect on interface gaps and sealing in four light-cured restorative materials using an original confocal microscopic methodology. The materials tested were an ormocer (Admira [Voco]), a compomer (Dyract AP [Dentsply]), a hybrid composite (Spectrum [Dentsply]), and a nanohybrid composite (Esthet·X [Dentsply]). Water sorption was evaluated by weighing material disks after immersion. Hygroscopic expansion was measured from volumetric variations of material fillings in cylindrical cavities in dentin slices; the interfacial gap size was obtained from the same cavities using a novel confocal microscopic method. Microleakage was evaluated in cavities prepared in extracted third molars. Measurements followed water immersion for 24 h, 1 week, 4 weeks, and 8 weeks. A factorial ANOVA, the Student Newman Keuls test for post-hoc comparisons, the Student's t-test, and the Pearson test were used for the statistical analysis (p hygroscopic expansion, and sealing. Hygroscopic expansion reduced post-polymerization interfacial gaps and improved cavity sealing. Dyract AP and Admira showed the highest water sorption, hygroscopic expansion, and gap size reduction. 1. The proposed methodology is valid to measure hygroscopic expansion and interfacial gap. 2. Water sorption and hygroscopic expansion are positively correlated, and hygroscopic expansion, gap size, and sealing are also positively correlated. 3. The adhesive influences the interfacial gap size and its variation after hygroscopic expansion. 4. Hygroscopic expansion reduces the interfacial gaps generated by polymerization shrinkage and improves cavity sealing.

  12. Cross-Sectional Shape of Rat Mesenteric Arterioles at Branching Studied by Confocal Laser Microscopy

    Science.gov (United States)

    Nakano, Atushi; Minamiyama, Motomu; Niimi, Hideyuki

    This study was aimed to investigate the cross-sectional shape of mesenteric arterioles at branching, using confocal laser microscopy. Wistar rats (8 weeks, male) were anesthetized with thiobutabarbital sodium. Blood flow and microvascular network in the mesentery were observed using video microscopy. The rat intestine with mesentery was extracted and the intestinal vasculature was perfused with Krebs-Ringer and then fixed with paraformaldehyde under a static pressure of 100mmHg. A section of mesentery was isolated from the intestine, and spread up to the in vivo geometry based on the intravital microscopic observation. The mesentery section was stained with tetramethyl rhodamine isothiocyanate (TRITC)-phalloidin. The samples were observed under a confocal laser microscope. The cross-sectional image was re-sliced to measure the cross-sectional area and major/minor axes of the best fitting ellipse. The aspect ratio was defined in terms of the minor/major diameter ratio. The extended focus image of mesenteric arterioles showed that the cross-sectional shape was not circular but elliptic-like. The cross-sectional area of the parent vessel decreased from proximal to distal positions. The mean aspect ratio of the parent vessel was approximately 0.5, while that of the branching vessel was approximately 0.8. The flattened shape and variation of the cross-sectional area of arterioles requires some correction of in vivo data of the two-dimensional mesenteric microvasculature obtained using intravital microscopy.

  13. Scanning a microhabitat: plant-microbe interactions revealed by confocal laser microscopy

    Directory of Open Access Journals (Sweden)

    Massimiliano eCardinale

    2014-03-01

    Full Text Available No plant or cryptogam exists in nature without microorganisms associated with its tissues. Plants as microbial hosts are puzzles of different microhabitats, each of them colonized by specifically adapted microbiomes. The interactions with such microorganisms have drastic effects on the host fitness. Since the last 20 years, the combination of microscopic tools and molecular approaches contributed to new insights into microbe-host interactions. Particularly, confocal laser scanning microscopy (CLSM facilitated the exploration of microbial habitats and allowed the observation of host-associated microorganisms in situ with an unprecedented accuracy. Here I present an overview of the progresses made in the study of the interactions between microorganisms and plants or plant-like organisms, focusing on the role of CLSM for the understanding of their significance. I critically discuss risks of misinterpretation when procedures of CLSM are not properly optimized. I also review approaches for quantitative and statistical analyses of CLSM images, the combination with other molecular and microscopic methods, and suggest the re-evaluation of natural autofluorescence. In this review, technical aspects were coupled with scientific outcomes, to facilitate the readers in identifying possible CLSM applications in their research or to expand their existing potential. The scope of this review is to highlight the importance of confocal microscopy in the study of plant-microbe interactions and also to be an inspiration for integrating microscopy with molecular techniques in future researches of microbial ecology.

  14. Impression Cytology in Eyes with Clinical and Confocal Scan Features of Acanthamoeba Keratitis

    Directory of Open Access Journals (Sweden)

    Mozhgan Rezaei Kanavi

    2013-01-01

    Full Text Available Purpose: To report impression cytology findings in specimens obtained from eyes with clinical and confocal microscopic features of Acanthamoeba keratitis (AK. Methods: In this interventional case series, impression cytology was obtained from corneas of patients with clinical and confocal microscopic features indicative of AK. Specimens were stained with Periodic acid-Schiff/Papanicolaou (PAS/PAP and examined for the presence of PAS-reactive Acanthamoeba cysts and/or hyperchromatic pear-shaped trophozoites. All specimens were then decolorized and re-stained with calcofluor white (CFW for the presence of chemofluorescent cysts. Results: Fifty-six eyes of 50 patients with mean age of 25.5±9.3 (range, 17 to 78 years were evaluated. Forty-one (82% cases were female and 51 (91.1% eyes had history of contact lens wear. PAS-reactive Acanthamoeba cysts and/or hyperchromatic pear-shaped trophozoites were identified in 53 eyes (94.6%, 2 of which demonstrated only trophozoitelike structures. CFW staining was able to reveal the presence of chemofluorescent cysts in all 51 specimens (91.1% in which cysts had been demonstrated with PAS/PAP staining. Trophozoites were not detected with CFW due to background staining of the cellulose acetate strip used for impression cytology. Conclusion: Corneal impression cytology, stained with PAS/PAP or with CFW, successfully detects Acanthamoeba and can be employed for early noninvasive diagnosis of AK.

  15. Adipocyte size and cellular expression of caveolar proteins analyzed by confocal microscopy

    DEFF Research Database (Denmark)

    Hulstrøm, Veronica; Prats Gavalda, Clara; Vinten, Jørgen

    2013-01-01

    Caveolae are abundant in adipocytes and are involved in the regulation of lipid accumulation, which is the main volume determinant of these cells. We have developed and applied a confocal microscopic technique for measuring individual cellular expression of the caveolar proteins cavin-1 and caveo......Caveolae are abundant in adipocytes and are involved in the regulation of lipid accumulation, which is the main volume determinant of these cells. We have developed and applied a confocal microscopic technique for measuring individual cellular expression of the caveolar proteins cavin-1...... and caveolin-1 along with the size of individual adipocytes. The technique was applied on collagenase isolated adipocytes from ad libitum fed Sprague-Dawley rats of different age (4-26 wk) and weight (103-629 g). We found that cellular expression of caveolar proteins was variable (SD of log expression...... in the range from 0.25 to 0.65). Regression analysis of protein expression on adipocyte size revealed that the expression of the caveolar proteins cavin-1 and caveolin-1 on adipocytes from individual rats was tightly related to adipocyte cell surface area (mean coefficient of regression was 0.83 for cavin...

  16. Assessment of Petrological Microscopes.

    Science.gov (United States)

    Mathison, Charter Innes

    1990-01-01

    Presented is a set of procedures designed to check the design, ergonomics, illumination, function, optics, accessory equipment, and image quality of a microscope being considered for purchase. Functions for use in a petrology or mineralogy laboratory are stressed. (CW)

  17. Scanning Auger Electron Microscope

    Data.gov (United States)

    Federal Laboratory Consortium — A JEOL model 7830F field emission source, scanning Auger microscope.Specifications / Capabilities:Ultra-high vacuum (UHV), electron gun range from 0.1 kV to 25 kV,...

  18. 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

  19. Three-dimensional reconstructions of solid surfaces using conventional microscopes.

    Science.gov (United States)

    Ficker, Tomáš; Martišek, Dalibor

    2016-01-01

    The three-dimensional digital replicas of solid surfaces are subject of interest of different branches of science and technology. The present paper in its introductory parts brings an overview of the various microscopic reconstructive techniques based on optical sectioning. The main attention is devoted to conventional reconstruction methods and especially to that one employing the Fourier transform. The three-dimensional replicas of this special reconstructive frequency method are compared graphically and numerically with the three-dimensional replicas of the confocal method. Based on the comparative study it has been concluded that the quality of the conventional replicas of surfaces possessing textures of intermediate height irregularities is acceptable and almost comparable with the quality of confocal replicas. This study is relevant both for identifying a convenient technique that provides good qualities of three-dimensional replicas and for selecting the hardware whose price is affordable even for small research groups studying rougher surface textures. © Wiley Periodicals, Inc.

  20. Minimizing inter-microscope variability in dental microwear texture analysis

    Science.gov (United States)

    Arman, Samuel D.; Ungar, Peter S.; Brown, Christopher A.; DeSantis, Larisa R. G.; Schmidt, Christopher; Prideaux, Gavin J.

    2016-06-01

    A common approach to dental microwear texture analysis (DMTA) uses confocal profilometry in concert with scale-sensitive fractal analysis to help understand the diets of extinct mammals. One of the main benefits of DMTA over other methods is the repeatable, objective manner of data collection. This repeatability, however, is threatened by variation in results of DMTA of the same dental surfaces yielded by different microscopes. Here we compare DMTA data of five species of kangaroos measured on seven profilers of varying specifications. Comparison between microscopes confirms that inter-microscope differences are present, but we show that deployment of a number of automated treatments to remove measurement noise can help minimize inter-microscope differences. Applying these same treatments to a published hominin DMTA dataset shows that they alter some significant differences between dietary groups. Minimising microscope variability while maintaining interspecific dietary differences requires then that these factors are balanced in determining appropriate treatments. The process outlined here offers a solution for allowing comparison of data between microscopes, which is essential for ongoing DMTA research. In addition, the process undertaken, including considerations of other elements of DMTA protocols also promises to streamline methodology, remove measurement noise and in doing so, optimize recovery of a reliable dietary signature.

  1. Deep stroma investigation by confocal microscopy

    Science.gov (United States)

    Rossi, Francesca; Tatini, Francesca; Pini, Roberto; Valente, Paola; Ardia, Roberta; Buzzonetti, Luca; Canovetti, Annalisa; Malandrini, Alex; Lenzetti, Ivo; Menabuoni, Luca

    2015-03-01

    Laser assisted keratoplasty is nowadays largely used to perform minimally invasive surgery and partial thickness keratoplasty [1-3]. The use of the femtosecond laser enables to perform a customized surgery, solving the specific problem of the single patient, designing new graft profiles and partial thickness keratoplasty (PTK). The common characteristics of the PTKs and that make them eligible respect to the standard penetrating keratoplasty, are: the preservation of eyeball integrity, a reduced risk of graft rejection, a controlled postoperative astigmatism. On the other hand, the optimal surgical results after these PTKs are related to a correct comprehension of the deep stroma layers morphology, which can help in the identification of the correct cleavage plane during surgeries. In the last years some studies were published, giving new insights about the posterior stroma morphology in adult subjects [4,5]. In this work we present a study performed on two groups of tissues: one group is from 20 adult subjects aged 59 +/- 18 y.o., and the other group is from 15 young subjects, aged 12+/-5 y.o.. The samples were from tissues not suitable for transplant in patients. Confocal microscopy and Environmental Scanning Electron Microscopy (ESEM) were used for the analysis of the deep stroma. The preliminary results of this analysis show the main differences in between young and adult tissues, enabling to improve the knowledge of the morphology and of the biomechanical properties of human cornea, in order to improve the surgical results in partial thickness keratoplasty.

  2. 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.

  3. 'En face' ex vivo reflectance confocal microscopy to help the surgery of basal cell carcinoma of the eyelid.

    Science.gov (United States)

    Espinasse, Marine; Cinotti, Elisa; Grivet, Damien; Labeille, Bruno; Prade, Virginie; Douchet, Catherine; Cambazard, Frédéric; Thuret, Gilles; Gain, Philippe; Perrot, Jean Luc

    2017-07-01

    Ex vivo confocal microscopy is a recent imaging technique for the perioperative control of skin tumour margins. Up to date, it has been used in the fluorescence mode and with vertical sections of the specimen margins. The aim of this study was to evaluate its use in the reflectance mode and with a horizontal ('en face') scanning of the surgical specimen in a series of basal cell carcinoma of the eyelid. Prospective consecutive cohort study was performed at the University Hospital of Saint-Etienne, France. Forty-one patients with 42 basal cell carcinoma of the eyelid participated in this study. Basal cell carcinomas were excised with a 2-mm-wide clinically safe margin. The surgical specimens were analysed under ex vivo confocal microscopy in the reflectance mode and with an en face scanning in order to control at a microscopic level if the margins were free from tumour invasion. Histopathogical examination was later performed in order to compare the results. Sensitivity and specificity of ex vivo confocal microscopy for the presence of tumour-free margins. Ex vivo confocal microscopy results were consistent with histopathology in all cases (tumour-free margins in 40 out of 42 samples; sensitivity and specificity of 100%). Ex vivo confocal microscopy in the reflectance mode with an 'en face' scanning can control tumour margins of eyelid basal cell carcinomas and optimize their surgical management. This procedure has the advantage on the fluorescent mode of not needing any contrast agent to examine the samples. © 2016 Royal Australian and New Zealand College of Ophthalmologists.

  4. Electron microscope studies

    Energy Technology Data Exchange (ETDEWEB)

    Crewe, A.V.; Kapp, O.H.

    1992-07-01

    This is a report covering the research performed in the Crewe laboratory between 1964 and 1992. Because of limitations of space we have provided relatively brief summaries of the major research directions of the facility during these years. A complete bibliography has been included and we have referenced groups of pertinent publications at the beginning of each section. This report summarizes our efforts to develop better electron microscopes and chronicles many of the experimental programs, in materials science and biology, that acted both as a stimulus to better microscope design and also as a testing ground for many instrumental innovations.

  5. Electron microscope studies

    International Nuclear Information System (INIS)

    Crewe, A.V.; Kapp, O.H.

    1992-01-01

    This is a report covering the research performed in the Crewe laboratory between 1964 and 1992. Because of limitations of space we have provided relatively brief summaries of the major research directions of the facility during these years. A complete bibliography has been included and we have referenced groups of pertinent publications at the beginning of each section. This report summarizes our efforts to develop better electron microscopes and chronicles many of the experimental programs, in materials science and biology, that acted both as a stimulus to better microscope design and also as a testing ground for many instrumental innovations

  6. Reconstruction of Axial Tomographic High Resolution Data from Confocal Fluorescence Microscopy: A Method for Improving 3D FISH Images

    Directory of Open Access Journals (Sweden)

    R. Heintzmann

    2000-01-01

    Full Text Available Fluorescent confocal laser scanning microscopy allows an improved imaging of microscopic objects in three dimensions. However, the resolution along the axial direction is three times worse than the resolution in lateral directions. A method to overcome this axial limitation is tilting the object under the microscope, in a way that the direction of the optical axis points into different directions relative to the sample. A new technique for a simultaneous reconstruction from a number of such axial tomographic confocal data sets was developed and used for high resolution reconstruction of 3D‐data both from experimental and virtual microscopic data sets. The reconstructed images have a highly improved 3D resolution, which is comparable to the lateral resolution of a single deconvolved data set. Axial tomographic imaging in combination with simultaneous data reconstruction also opens the possibility for a more precise quantification of 3D data. The color images of this publication can be accessed from http://www.esacp.org/acp/2000/20‐1/heintzmann.htm. At this web address an interactive 3D viewer is additionally provided for browsing the 3D data. This java applet displays three orthogonal slices of the data set which are dynamically updated by user mouse clicks or keystrokes.

  7. 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.

  8. Diagnostic value of the confocal laser scanning microscopy in vivo

    Directory of Open Access Journals (Sweden)

    A. A. Kubanova

    2015-01-01

    Full Text Available The confocal laser scanning microscopy in vivo is a promising study method to visualize cell structures of epidermis and papillary dermis without affecting the skin integrity, which provides for a resolution and contrast similar to those characteristic of the classical histology examination. Goal. To assess the confocal laser scanning microscopy in vivo (CLSM technique for diagnosing actinic keratosis, psoriasis vulgaris and rosacea vs. the classical histology examination. Study materials. The article describes the results obtained by using the confocal laser scanning microscopy in vivo technique vs. the histology examination in 10 patients with erythematous actinic keratosis, 10 patients with extensive psoriasis and 10 patients with erythematous and papulous rosacea. Results. The article describes diagnostically significant signs of the diseases detected by using the confocal laser scanning microscopy in vivo as well as the potential of this method in terms of diagnosing inflammatory skin diseases.

  9. Confocal optical system: a novel noninvasive sensor to study mixing.

    Science.gov (United States)

    Vallejos, Jose R; Kostov, Yordan; Marten, Mark R; Rao, Govind

    2005-01-01

    A novel confocal optical system to study mixing time in small-scale bioreactors is presented. The system is designed to monitor fluorescence upon tracer addition from a localized confocal volume of 0.21 mL within a glass vessel. The key elements of the fluorescence-based confocal system are a pinhole, a lens, an APD (Avalanche photodiode) detector, and light filters. The optical technique was validated by comparison with a pH-based technique. Finally, the optical sensor was tested and a real cultivation media (i.e., spent mammalian cell media) was used to measure mixing time in a 12.5-mL stirred transparent vessel. High accuracy, easy results interpretation, and low costs are the three most attractive characteristics of the sensor. Because of its noninvasive nature and versatility, the results suggest that the confocal system is a promising tool to perform mixing time studies in stirred vessels.

  10. [Diagnostic value of confocal microscopy in primary corneal ectasia].

    Science.gov (United States)

    Egorova, G B; Rogova, A Ia; Mitichkina, T S

    2012-01-01

    Diagnostic value of confocal microscopy in subclinical corneal ectasia is analysed. Morphological changes were revealed in 71.2% of cases with subclinical keratoconus. Results of the study let us reveal typical morphological abnormalities in subclinical keratoconus, although these changes are not specific for this condition. The method is also not informative for detecting of early signs of pellucid marginal degeneration. Confocal microscopy may be considered an additional specifying method in diagnostic algorithm of subclinical corneal ectasia.

  11. Cuticle and subsurface ornamentation of intact plant leaf epidermis under confocal and superresolution microscopy.

    Science.gov (United States)

    Urban, Michael A; Barclay, Richard S; Sivaguru, Mayandi; Punyasena, Surangi W

    2018-02-01

    Plant cuticle micromorphology is an invaluable tool in modern ecology and paleoecology. It has expanded our knowledge of systematic relationships among diverse plant groups and can be used to identify fossil plants. Furthermore, fossil plant leaf micromorphology is used for reconstructing past environments, most notably for estimating atmospheric CO 2 concentration. Here we outline a new protocol for imaging plant cuticle for archival and paleoecological applications. Traditionally, both modern reference and fossil samples undergo maceration with subsequent imaging via environmental SEM, widefield fluorescence, or light microscopy. In this paper, we demonstrate the capabilities of alternative preparation and imaging methods using confocal and superresolution microscopy with intact leaf samples. This method produces detailed three-dimensional images of surficial and subsurface structures of the intact leaf. Multiple layers are captured simultaneously, which previously required independent maceration and microtome steps. We compared clearing agents (chloral hydrate, KOH, and Visikol); mounting media (Eukitt and Hoyer's); fluorescent stains (periodic acid Schiff, propidium iodide); and confocal vs. superresolution microscopes. We conclude that Eukitt is the best medium for long-term preservation and imaging. Because of nontoxicity and ease of procurement, Visikol made for the best clearing agent. Staining improves contrast and under most circumstances PAS provided the clearest images. Supperresolution produced higher clarity images than traditional confocal, but the information gained was minimal. This new protocol provides the botanical and paleobotanical community an alternative to traditional techniques. Our proposed workflow has the net benefit of being more efficient than traditional methods, which only capture the surface of the plant epidermis. Microsc. Res. Tech. 81:129-140, 2018. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  12. Sensitivity and Specificity of Cardiac Tissue Discrimination Using Fiber-Optics Confocal Microscopy.

    Science.gov (United States)

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

    2016-01-01

    Disturbances of the cardiac conduction system constitute a major risk after surgical repair of complex cases of congenital heart disease. Intraoperative identification of the conduction system may reduce the incidence of these disturbances. We previously developed an approach to identify cardiac tissue types using fiber-optics confocal microscopy and extracellular fluorophores. Here, we applied this approach to investigate sensitivity and specificity of human and automated classification in discriminating images of atrial working myocardium and specialized tissue of the conduction system. Two-dimensional image sequences from atrial working myocardium and nodal tissue of isolated perfused rodent hearts were acquired using a fiber-optics confocal microscope (Leica FCM1000). We compared two methods for local application of extracellular fluorophores: topical via pipette and with a dye carrier. Eight blinded examiners evaluated 162 randomly selected images of atrial working myocardium (n = 81) and nodal tissue (n = 81). In addition, we evaluated the images using automated classification. Blinded examiners achieved a sensitivity and specificity of 99.2 ± 0.3% and 98.0 ± 0.7%, respectively, with the dye carrier method of dye application. Sensitivity and specificity was similar for dye application via a pipette (99.2 ± 0.3% and 94.0 ± 2.4%, respectively). Sensitivity and specificity for automated methods of tissue discrimination were similarly high. Human and automated classification achieved high sensitivity and specificity in discriminating atrial working myocardium and nodal tissue. We suggest that our findings facilitate clinical translation of fiber-optics confocal microscopy as an intraoperative imaging modality to reduce the incidence of conduction disturbances during surgical correction of congenital heart disease.

  13. Quantitative analysis of microbicide concentrations in fluids, gels and tissues using confocal Raman spectroscopy.

    Directory of Open Access Journals (Sweden)

    Oranat Chuchuen

    Full Text Available Topical vaginal anti-HIV microbicides are an important focus in female-based strategies to prevent the sexual transmission of HIV. Understanding microbicide pharmacokinetics is essential to development, characterization and implementation of efficacious microbicide drug delivery formulations. Current methods to measure drug concentrations in tissue (e.g., LC-MS/MS, liquid chromatography coupled with tandem mass spectrometry are highly sensitive, but destructive and complex. This project explored the use of confocal Raman spectroscopy to detect microbicide drugs and to measure their local concentrations in fluids, drug delivery gels, and tissues. We evaluated three candidate microbicide drugs: tenofovir, Dapivirine and IQP-0528. Measurements were performed in freshly excised porcine buccal tissue specimens, gel vehicles and fluids using two Horiba Raman microscopes, one of which is confocal. Characteristic spectral peak calibrations for each drug were obtained using serial dilutions in the three matrices. These specific Raman bands demonstrated strong linear concentration dependences in the matrices and were characterized with respect to their unique vibrational signatures. At least one specific Raman feature was identified for each drug as a marker band for detection in tissue. Sensitivity of detection was evaluated in the three matrices. A specific peak was also identified for tenofovir diphosphate, the anti-HIV bioactive product of tenofovir after phosphorylation in host cells. Z-scans of drug concentrations vs. depth in excised tissue specimens, incubated under layers of tenofovir solution in a Transwell assay, showed decreasing concentration with depth from the surface into the tissue. Time-dependent concentration profiles were obtained from tissue samples incubated in the Transwell assay, for times ranging 30 minutes - 6 hours. Calibrations and measurements from tissue permeation studies for tenofovir showed good correlation with gold

  14. Scanning transmission electron microscope

    NARCIS (Netherlands)

    Kruit, P.

    2006-01-01

    The invention relates to a scanning transmission electron microscope comprising an electron source, an electron accelerator and deflection means for directing electrons emitted by the electron source at an object to be examined, and in addition a detector for detecting electrons coming from the

  15. MICROSCOPIC, PHYSICOCHEMICAL AND CHROMATOGRAPHIC ...

    African Journals Online (AJOL)

    Peters

    MICROSCOPIC, PHYSICOCHEMICAL AND CHROMATOGRAPHIC. FINGERPRINTS OF LEAVES OF NIGERIAN CASSIA TORA LINN. Fatokun Omolola T1*., EsievoKevwe B2., Ugbabe Grace E3. and Kunle Oluyemisi F4. Department of Medicinal Plant Research and Traditional Medicine, National Institute for.

  16. SPM: Scanning positron microscope

    Directory of Open Access Journals (Sweden)

    Marcel Dickmann

    2015-08-01

    Full Text Available The Munich scanning positron microscope, operated by the Universität der Bundeswehr München and the Technische Universität München, located at NEPOMUC, permits positron lifetime measurements with a lateral resolution in the µm range and within an energy range of 1 – 20 keV.

  17. Microscope on Mars

    Science.gov (United States)

    2004-01-01

    This image taken at Meridiani Planum, Mars by the panoramic camera on the Mars Exploration Rover Opportunity shows the rover's microscopic imager (circular device in center), located on its instrument deployment device, or 'arm.' The image was acquired on the ninth martian day or sol of the rover's mission.

  18. Making Art with Microscopes

    Science.gov (United States)

    Benedis-Grab, Gregory

    2011-01-01

    Interdisciplinary teaching is a great way to focus on overarching concepts and help students make connections across disciplines. Historically, art and science have been connected disciplines. The botanical prints of the 18th and 19th centuries and early work with microscopes are two examples of a need for strong artistic skills in the science…

  19. Terahertz scanning probe microscope

    NARCIS (Netherlands)

    Klapwijk, T.M.

    2014-01-01

    The invention provides aterahertz scanning probe microscope setup comprising (i) a terahertz radiation source configured to generate terahertz radiation; (ii) a terahertz lens configured to receive at least part of the terahertz radiation from the terahertz radiation source; (iii) a cantilever unit

  20. Direct in situ viability assessment of bacteria in probiotic dairy products using viability staining in conjunction with confocal scanning laser microscopy.

    Science.gov (United States)

    Auty, M A; Gardiner, G E; McBrearty, S J; O'Sullivan, E O; Mulvihill, D M; Collins, J K; Fitzgerald, G F; Stanton, C; Ross, R P

    2001-01-01

    The viability of the human probiotic strains Lactobacillus paracasei NFBC 338 and Bifidobacterium sp. strain UCC 35612 in reconstituted skim milk was assessed by confocal scanning laser microscopy using the LIVE/DEAD BacLight viability stain. The technique was rapid (milk and fermented milk. These results indicate the value of the microscopic approach for rapid viability testing of such probiotic products. In contrast, the numbers obtained by direct microscopic counting for Cheddar cheese and spray-dried probiotic milk powder were lower than those obtained by plate counting. These results highlight the limitations of LIVE/DEAD BacLight staining and the need to optimize the technique for different strain-product combinations. The minimum detection limit for in situ viability staining in conjunction with confocal scanning laser microscopy enumeration was approximately 10(8) bacteria/ml (equivalent to approximately 10(7) CFU/ml), based on Bifidobacterium sp. strain UCC 35612 counts in maximum-recovery diluent.

  1. Improved sampling and analysis of images in corneal confocal microscopy.

    Science.gov (United States)

    Schaldemose, E L; Fontain, F I; Karlsson, P; Nyengaard, J R

    2017-10-01

    Corneal confocal microscopy (CCM) is a noninvasive clinical method to analyse and quantify corneal nerve fibres in vivo. Although the CCM technique is in constant progress, there are methodological limitations in terms of sampling of images and objectivity of the nerve quantification. The aim of this study was to present a randomized sampling method of the CCM images and to develop an adjusted area-dependent image analysis. Furthermore, a manual nerve fibre analysis method was compared to a fully automated method. 23 idiopathic small-fibre neuropathy patients were investigated using CCM. Corneal nerve fibre length density (CNFL) and corneal nerve fibre branch density (CNBD) were determined in both a manual and automatic manner. Differences in CNFL and CNBD between (1) the randomized and the most common sampling method, (2) the adjusted and the unadjusted area and (3) the manual and automated quantification method were investigated. The CNFL values were significantly lower when using the randomized sampling method compared to the most common method (p = 0.01). There was not a statistical significant difference in the CNBD values between the randomized and the most common sampling method (p = 0.85). CNFL and CNBD values were increased when using the adjusted area compared to the standard area. Additionally, the study found a significant increase in the CNFL and CNBD values when using the manual method compared to the automatic method (p ≤ 0.001). The study demonstrated a significant difference in the CNFL values between the randomized and common sampling method indicating the importance of clear guidelines for the image sampling. The increase in CNFL and CNBD values when using the adjusted cornea area is not surprising. The observed increases in both CNFL and CNBD values when using the manual method of nerve quantification compared to the automatic method are consistent with earlier findings. This study underlines the importance of improving the analysis of the

  2. 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

  3. [Antiphospholipid antibodies and recurrent abortions: possible pathogenetic role of annexin A5 investigated by confocal microscopy].

    Science.gov (United States)

    Peluso, G; Morrone, G

    2007-06-01

    It has been established that antiphospholipid antibodies (aPL) are associated with recurrent abortions, but the pathophysiologic mechanisms that characterize thrombosis and recurrent pregnancy losses are still not clear.However, it is known that they are associated with the presence of antibodies directed against anionic phospholipids and putative cofactors. In this study the pathogenetic role of annexin A5, a potent anticoagulant cofactor protein for its anticoagulant property in recurrent abortions, was investigated. Endothelial cells of human umbilical veins ''EAHY2936 Line'' in culture were used, incubated with antiphospholipid anticardiolipin (ACA) antibodies purified from plasma of patients with recurrent abortions. The expression of annexin A5 on the cells with ACA was investigated by immunofluorescence and by confocal microscope. The negative control was also carried out: EAHY cells in cultivation medium without ACA. Confocal analysis revealed a uniform distribution of annexin A5 on the cellular membranes in the negative control. Instead, in EAHY cells with ACA, the annexin A5 appears distributed in irregular manners on the cellular membranes (cytoplasmic and nuclear). The results of an irregular ''cluster'' distribution of annexin A5 on the EAHY cells in presence of aPL, and in agreement with the literature, demonstrated that aPL, inhibiting annexin A5 ability to protect anionic phospholipid, promote the coagulation factors to diffuse laterally against phospholipids.

  4. Monitoring treatment response in psoriasis: current perspectives on the clinical utility of reflectance confocal microscopy

    Directory of Open Access Journals (Sweden)

    Agozzino M

    2017-02-01

    Full Text Available Marina Agozzino,1 Cecilia Noal,2 Francesco Lacarrubba,3 Marco Ardigò4 1Dermatology Unit, Second University of Naples, Naples, 2Dermatology Unit, University of Trieste, Trieste, 3Dermatology Clinic, University of Catania, Catania, 4Clinical Dermatology Department, San Gallicano Dermatological Institute, Istituto di Ricovero e Cura a Carattere Scientifico, Rome, Italy Abstract: Reflectance confocal microscopy (RCM evaluation of inflammatory skin diseases represents a relatively new technique that, during the past 5 years, has attracted increasing interest, with consequent progressive increment of publications in literature. The success of RCM is directly related to the high need for noninvasive techniques able to both reduce the number of skin biopsies and support clinical diagnosis and patient management. RCM helps to visualize microscopic descriptors of plaque psoriasis (PP with good reproducibility between observers and a high grade of correspondence with histopathology. Several clinical tests are used for the therapeutic management of PP, but they are limited by subjective interpretation. Skin biopsy presents objective interpretation, but the procedure is invasive and not repeatable. RCM has been used not only for the evaluation of skin cancer or inflammatory skin diseases, but also for monitoring the efficacy of different treatments in PP. In this review, we present some examples of RCM applications in therapeutic psoriasis follow-up. Keywords: reflectance confocal microscopy, psoriasis noninvasive follow-up, psoriasis monitoring

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

    Science.gov (United States)

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

    2012-03-01

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

  6. Morphological study of adult male worms of Schistosoma mansoni Sambon, 1907 by confocal laser scanning microscopy

    Directory of Open Access Journals (Sweden)

    Machado-Silva José Roberto

    1998-01-01

    Full Text Available Aiming to detail data obtained through brightfield microscopy (BM on reproductive, excretory and digestive system, specimens of Schistosoma mansoni eight weeks old, were recovered from SW mice, stained with Langeron's carmine and analyzed under a confocal laser scanning microscope CLSM 410 (Carl Zeiss. The reproductive system presented a single and lobate testis, with intercommunications between the lobes without efferent duct. Supernumerary testicular lobe was amorphous and isolated from the normal ones. Collecting tubules (excretory ducts, followed by the excretory bladder, opening to the external media through the excretory pore, were observed at the posterior extremity of the body. In the digestive tract, a cecal swelling was noted at the junction that originates the single cecum. It was concluded that through confocal laser scanning microscopy, new interpretations of morphological structures of S. mansoni worms could be achieved, modifying adopted and current descriptions. The gonad consists of a single lobed testis, similar to that observed in some trematode species. Moreover, the same specimens can be observed either by BM or CLSM, considering that the latter causes only focal and limited damage in tissue structures.

  7. Parameter-free binarization and skeletonization of fiber networks from confocal image stacks.

    Directory of Open Access Journals (Sweden)

    Patrick Krauss

    Full Text Available We present a method to reconstruct a disordered network of thin biopolymers, such as collagen gels, from three-dimensional (3D image stacks recorded with a confocal microscope. The method is based on a template matching algorithm that simultaneously performs a binarization and skeletonization of the network. The size and intensity pattern of the template is automatically adapted to the input data so that the method is scale invariant and generic. Furthermore, the template matching threshold is iteratively optimized to ensure that the final skeletonized network obeys a universal property of voxelized random line networks, namely, solid-phase voxels have most likely three solid-phase neighbors in a 3 x 3 x 3 neighborhood. This optimization criterion makes our method free of user-defined parameters and the output exceptionally robust against imaging noise.

  8. Live cell refractometry using Hilbert phase microscopy and confocal reflectance microscopy.

    Science.gov (United States)

    Lue, Niyom; Choi, Wonshik; Popescu, Gabriel; Yaqoob, Zahid; Badizadegan, Kamran; Dasari, Ramachandra R; Feld, Michael S

    2009-11-26

    Quantitative chemical analysis has served as a useful tool for understanding cellular metabolisms in biology. Among many physical properties used in chemical analysis, refractive index in particular has provided molecular concentration that is an important indicator for biological activities. In this report, we present a method of extracting full-field refractive index maps of live cells in their native states. We first record full-field optical thickness maps of living cells by Hilbert phase microscopy and then acquire physical thickness maps of the same cells using a custom-built confocal reflectance microscope. Full-field and axially averaged refractive index maps are acquired from the ratio of optical thickness to physical thickness. The accuracy of the axially averaged index measurement is 0.002. This approach can provide novel biological assays of label-free living cells in situ.

  9. Live Cell Refractometry Using Hilbert Phase Microscopy and Confocal Reflectance Microscopy†

    Science.gov (United States)

    Lue, Niyom; Choi, Wonshik; Popescu, Gabriel; Yaqoob, Zahid; Badizadegan, Kamran; Dasari, Ramachandra R.; Feld, Michael S.

    2010-01-01

    Quantitative chemical analysis has served as a useful tool for understanding cellular metabolisms in biology. Among many physical properties used in chemical analysis, refractive index in particular has provided molecular concentration that is an important indicator for biological activities. In this report, we present a method of extracting full-field refractive index maps of live cells in their native states. We first record full-field optical thickness maps of living cells by Hilbert phase microscopy and then acquire physical thickness maps of the same cells using a custom-built confocal reflectance microscope. Full-field and axially averaged refractive index maps are acquired from the ratio of optical thickness to physical thickness. The accuracy of the axially averaged index measurement is 0.002. This approach can provide novel biological assays of label-free living cells in situ. PMID:19803506

  10. Surface determination of 3D confocal Raman microscopy imaging of the skin

    Science.gov (United States)

    Schleusener, J.; Carrer, V.; Patzelt, A.; Lademann, J.; Darvin, M. E.

    2017-12-01

    A surface determination method for the application of 3D confocal Raman microscopy on inhomogeneous skin sections has been presented, which is based on depth profiles of the keratin contribution of the acquired Raman spectra. The method was compared to two similar auto-focusing methods that are based on the intensity of the reflected excitation light and Raman spectra, respectively. The measurements were performed on hair follicles containing skin sections of porcine ears ex vivo. The surface determination on such samples is especially challenging due to their different molecular composition and surface inhomogeneity. An advantage of this method is molecular sensitivity, whereby only the surface of the sample will be detected and not the substrate of the microscope slide, in the case of disruptions during the processing of samples. A disadvantage of the method is the increased overall acquisition time if only the surface spectra are to be applied for 2D mapping.

  11. Electric field and energy of a point electric charge between confocal hyperbolaidal electrodes

    Energy Technology Data Exchange (ETDEWEB)

    Ley-Koo, E. [Universidad Nacional Autonoma de Mexico, Mexico, D. F. (Mexico)

    2001-06-01

    The electric potential and intensity field, as well as the energy of a point electric charge between confocal hyperboloidal electrodes is evaluated as a superposition of prolate spheroidal harmonics using the Green-function technique. This study is motivated by the need to model the electric field between the tip and the sample in a scanning tunnelling microscope, and it can also be applied to a conductor-insulator-conductor junction. [Spanish] Los campos de potencial y de intensidad electrica, asi como la energia de una carga electrica puntual entre electrodos hiperboloidales confocales se evaluan como superposiciones de armonicos esferoidales prolatos usando la tecnica de la funcion de Green. Este estudio ha sido motivado por la necesidad de modelar el campo electrico entre la punta y la muestra de un microscopio de tunelamiento y barrido, y se puede aplicar tambien a una union de conductor-aislante-conductor.

  12. 3D-confocal microscopy for surface analysis of microstructured materials

    Science.gov (United States)

    Kagerer, Bernd; Brodmann, Rainer; Valentin, Juergen; Filzek, Jan; Popp, Uwe

    2002-06-01

    The surface of technical materials is playing an ever more important part in modern production processes. However, standard roughness values, which are obtained from a profile, frequently no longer provide sufficient descriptions. What are desired are three-dimensional measurements of surfaces over a macroscopic range with a high degree of vertical and lateral resolution. This has become necessary to be able to describe both deterministic and non-deterministic structures in the same fashion. Due to increased requirements for data and the measuring speed demanded by industry, only optical systems are a possibility. Using the example of tribology, the capability of this technology is shown in this article on the basis of the commercial confocal 3D white light microscope, the NanoFocusTMμSurfTM. On the one hand, the technology and data preparation used are discussed, and on the other, a comparison is drawn with other standard optical measuring methods.

  13. Detection of living Sarcoptes scabiei larvae by reflectance mode confocal microscopy in the skin of a patient with crusted scabies

    Science.gov (United States)

    Levi, Assi; Mumcuoglu, Kosta Y.; Ingber, Arieh; Enk, Claes D.

    2012-06-01

    Scabies is an intensely pruritic disorder induced by a delayed type hypersensitivity reaction to infestation of the skin by the mite Sarcoptes scabiei. The diagnosis of scabies is established clinically and confirmed by identifying mites or eggs by microscopic examination of scrapings from the skin or by surface microscopy using a dermatoscope. Reflectance-mode confocal microscopy is a novel technique used for noninvasive imaging of skin structures and lesions at a resolution compatible to that of conventional histology. Recently, the technique was employed for the confirmation of the clinical diagnosis of scabies. We demonstrate the first ever documentation of a larva moving freely inside the skin of a patient infected with scabies.

  14. Monitoring the dynamics of phase separation in a polymer blend by confocal imaging and fluorescence correlation spectroscopy.

    Science.gov (United States)

    Doroshenko, Mikheil; Gonzales, Maria; Best, Andreas; Butt, Hans-Jürgen; Koynov, Kaloian; Floudas, George

    2012-09-26

    The phase separation of the polymer blend polystyrene/poly(methyl phenyl siloxane) (PS/PMPS) is studied in situ by laser scanning confocal microscopy (LSCM) and by fluorescence correlation spectroscopy (FCS) at macroscopic and microscopic length scales, respectively. It is shown for the first time that FCS when combined with LSCM can provide independent information on the local concentration within the phase-separated domains as well as the interfacial width. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Confocal Microscopy on Light-emitting Nanostructures and X-ray Imaging Detectors Based on Color Centers in Lithium Fluoride

    International Nuclear Information System (INIS)

    Bonfigli, F.; Almaviva, S.; Montereali, R. M.

    2010-01-01

    Confocal Laser Scanning Microscope (CLSM) is a versatile and powerful optical instrument which is gaining a strong increase of interest for biological investigations and also for the characterization of materials, microstructures and devices. We exploit its capability for the characterization of light-emitting micro and nano-structures based on color centers in lithium fluoride. CLSM was successfully used as an advanced optical reading system to detect X-ray micro-radiographies of biological specimens stored in LiF imaging detectors.

  16. Confocal microscopy assessment of filling material remaining on root canal walls after retreatment.

    Science.gov (United States)

    Rached-Júnior, F A; Sousa-Neto, M D; Bruniera, J F B; Duarte, M A H; Silva-Sousa, Y T C

    2014-03-01

    The percentage of Endofill remaining on canal walls after retreatment with different techniques was evaluated using confocal microscopy and qualitative analysis of the interface between the filling material/dentine. Sixty-four root canals of incisors were prepared with ProTaper, filled with gutta-percha and Endofill mixed with 0.1% rhodamine B. The roots were thermocycled and distributed into groups according to the method of evaluation: GI - direct viewing (DV) and GII - operating microscope (OM) and according to the removal technique: A) ProTaper retreatment (PR), B) PR/xylol, C) ultrasound and D) ultrasound/xylol. The root canals were then refilled with gutta-percha and AH Plus with 0.1% fluorescein and sectioned at 2, 4 and 6 mm from the apex. The percentage of remaining Endofill was analysed by confocal microscopy. Additionally, 16 roots were prepared with a ProTaper F5 instrument and were filled with Endofill + 0.1% rhodamine B/gutta-percha (negative control group) (n = 8), and the positive control group (n = 8) were filled with AH Plus with 0.1% fluorescein/gutta-percha. Three-way anova demonstrated differences in the method of evaluation, removal techniques and their interaction (P  0.05). Qualitative analysis detected that ultrasound had lower remaining Endofill than PR, particularly when combined with xylol. Operating microscope resulted in lower residual sealer, regardless of the removal technique. None of the protocols was associated with complete removal of the filling material; however, the use of ultrasound/xylol under an OM provided better results. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  17. Improving axial resolution in confocal microscopy with new high refractive index mounting media.

    Directory of Open Access Journals (Sweden)

    Coralie Fouquet

    Full Text Available Resolution, high signal intensity and elevated signal to noise ratio (SNR are key issues for biologists who aim at studying the localisation of biological structures at the cellular and subcellular levels using confocal microscopy. The resolution required to separate sub-cellular biological structures is often near to the resolving power of the microscope. When optimally used, confocal microscopes may reach resolutions of 180 nm laterally and 500 nm axially, however, axial resolution in depth is often impaired by spherical aberration that may occur due to refractive index mismatches. Spherical aberration results in broadening of the point-spread function (PSF, a decrease in peak signal intensity when imaging in depth and a focal shift that leads to the distortion of the image along the z-axis and thus in a scaling error. In this study, we use the novel mounting medium CFM3 (Citifluor Ltd., UK with a refractive index of 1.518 to minimize the effects of spherical aberration. This mounting medium is compatible with most common fluorochromes and fluorescent proteins. We compare its performance with established mounting media, harbouring refractive indices below 1.500, by estimating lateral and axial resolution with sub-resolution fluorescent beads. We show furthermore that the use of the high refractive index media renders the tissue transparent and improves considerably the axial resolution and imaging depth in immuno-labelled or fluorescent protein labelled fixed mouse brain tissue. We thus propose to use those novel high refractive index mounting media, whenever optimal axial resolution is required.

  18. Intensity calibration and flat-field correction for fluorescence microscopes.

    Science.gov (United States)

    Model, Michael

    2014-04-01

    Standardization in fluorescence microscopy involves calibration of intensity in reproducible units and correction for spatial nonuniformity of illumination (flat-field or shading correction). Both goals can be achieved using concentrated solutions of fluorescent dyes. When a drop of a highly concentrated fluorescent dye is placed between a slide and a coverslip it produces a spatially uniform field, resistant to photobleaching and with reproducible quantum yield; it can be used as a brightness standard for wide-field and confocal microscopes. For wide-field microscopes, calibration can be further extended to absolute molecular units. This can be done by imaging a solution of known concentration and known depth; the latter can be prepared by placing a small spherical lens in a diluted solution of the same fluorophore that is used in the biological specimen. Copyright © 2014 John Wiley & Sons, Inc.

  19. Microscopic dynamical Casimir effect

    Science.gov (United States)

    Souza, Reinaldo de Melo e.; Impens, François; Neto, Paulo A. Maia

    2018-03-01

    We consider an atom in its ground state undergoing a nonrelativistic oscillation in free space. The interaction with the electromagnetic quantum vacuum leads to two effects to leading order in perturbation theory. When the mechanical frequency is larger than the atomic transition frequency, the dominant effect is the motion-induced transition to an excited state with the emission of a photon carrying the excess energy. We compute the angular distribution of emitted photons and the excitation rate. On the other hand, when the mechanical frequency is smaller than the transition frequency, the leading-order effect is the parametric emission of photon pairs, which constitutes the microscopic counterpart of the dynamical Casimir effect. We discuss the properties of the microscopic dynamical Casimir effect and build a connection with the photon production by an oscillating macroscopic metallic mirror.

  20. Microscopic approach to polaritons

    DEFF Research Database (Denmark)

    Skettrup, Torben

    1981-01-01

    The interaction between excitons and light has been investigated in detail. The perturbational approach turns out to be invalid. However, an exact solution can be obtained directly from the Schrödinger equation for a fixed light field. This solution corresponds to a nonlinear optical response...... contrary to experimental experience. In order to remove this absurdity the semiclassical approach must be abandoned and the electromagnetic field quantized. A simple microscopic polariton model is then derived. From this the wave function for the interacting exciton-photon complex is obtained...... of light of the crystal. The introduction of damping smears out the excitonic spectra. The wave function of the polariton, however, turns out to be very independent of damping up to large damping values. Finally, this simplified microscopic polariton model is compared with the exact solutions obtained...

  1. Ion photon emission microscope

    Science.gov (United States)

    Doyle, Barney L.

    2003-04-22

    An ion beam analysis system that creates microscopic multidimensional image maps of the effects of high energy ions from an unfocussed source upon a sample by correlating the exact entry point of an ion into a sample by projection imaging of the ion-induced photons emitted at that point with a signal from a detector that measures the interaction of that ion within the sample. The emitted photons are collected in the lens system of a conventional optical microscope, and projected on the image plane of a high resolution single photon position sensitive detector. Position signals from this photon detector are then correlated in time with electrical effects, including the malfunction of digital circuits, detected within the sample that were caused by the individual ion that created these photons initially.

  2. Neuromorphic Data Microscope

    Energy Technology Data Exchange (ETDEWEB)

    Naegle, John H.; Suppona, Roger A.; Aimone, James Bradley; James, Conrad D.; Follett, David R.; Townsend, Duncan C.M.; Follett, Pamela L.; Karpman, Gabe D.

    2017-08-01

    In 2016, Lewis Rhodes Labs, (LRL), shipped the first commercially viable Neuromorphic Processing Unit, (NPU), branded as a Neuromorphic Data Microscope (NDM). This product leverages architectural mechanisms derived from the sensory cortex of the human brain to efficiently implement pattern matching. LRL and Sandia National Labs have optimized this product for streaming analytics, and demonstrated a 1,000x power per operation reduction in an FPGA format. When reduced to an ASIC, the efficiency will improve to 1,000,000x. Additionally, the neuromorphic nature of the device gives it powerful computational attributes that are counterintuitive to those schooled in traditional von Neumann architectures. The Neuromorphic Data Microscope is the first of a broad class of brain-inspired, time domain processors that will profoundly alter the functionality and economics of data processing.

  3. Thermal Lens Microscope

    Science.gov (United States)

    Uchiyama, Kenji; Hibara, Akihide; Kimura, Hiroko; Sawada, Tsuguo; Kitamori, Takehiko

    2000-09-01

    We developed a novel laser microscope based on the thermal lens effect induced by a coaxial beam comprised of excitation and probe beams. The signal generation mechanism was confirmed to be an authentic thermal lens effect from the measurement of signal and phase dependences on optical configurations between the sample and the probe beam focus, and therefore, the thermal lens effect theory could be applied. Two-point spatial resolution was determined by the spot size of the excitation beam, not by the thermal diffusion length. Sensitivity was quite high, and the detection ability, evaluated using a submicron microparticle containing dye molecules, was 0.8 zmol/μm2, hence a distribution image of trace chemical species could be obtained quantitatively. In addition, analytes are not restricted to fluorescent species, therefore, the thermal lens microscope is a promising analytical microscope. A two-dimensional image of a histamine molecule distribution, which was produced in mast cells at the femtomole level in a human nasal mucous polyp, was obtained.

  4. Digital adaptive optics line-scanning confocal imaging system

    Science.gov (United States)

    Liu, Changgeng; Kim, Myung K.

    2015-01-01

    Abstract. A digital adaptive optics line-scanning confocal imaging (DAOLCI) system is proposed by applying digital holographic adaptive optics to a digital form of line-scanning confocal imaging system. In DAOLCI, each line scan is recorded by a digital hologram, which allows access to the complex optical field from one slice of the sample through digital holography. This complex optical field contains both the information of one slice of the sample and the optical aberration of the system, thus allowing us to compensate for the effect of the optical aberration, which can be sensed by a complex guide star hologram. After numerical aberration compensation, the corrected optical fields of a sequence of line scans are stitched into the final corrected confocal image. In DAOLCI, a numerical slit is applied to realize the confocality at the sensor end. The width of this slit can be adjusted to control the image contrast and speckle noise for scattering samples. DAOLCI dispenses with the hardware pieces, such as Shack–Hartmann wavefront sensor and deformable mirror, and the closed-loop feedbacks adopted in the conventional adaptive optics confocal imaging system, thus reducing the optomechanical complexity and cost. Numerical simulations and proof-of-principle experiments are presented that demonstrate the feasibility of this idea. PMID:26140334

  5. (LMRG): Microscope Resolution, Objective Quality, Spectral Accuracy and Spectral Un-mixing

    Science.gov (United States)

    Bayles, Carol J.; Cole, Richard W.; Eason, Brady; Girard, Anne-Marie; Jinadasa, Tushare; Martin, Karen; McNamara, George; Opansky, Cynthia; Schulz, Katherine; Thibault, Marc; Brown, Claire M.

    2012-01-01

    The second study by the LMRG focuses on measuring confocal laser scanning microscope (CLSM) resolution, objective lens quality, spectral imaging accuracy and spectral un-mixing. Affordable test samples for each aspect of the study were designed, prepared and sent to 116 labs from 23 countries across the globe. Detailed protocols were designed for the three tests and customized for most of the major confocal instruments being used by the study participants. One protocol developed for measuring resolution and objective quality was recently published in Nature Protocols (Cole, R. W., T. Jinadasa, et al. (2011). Nature Protocols 6(12): 1929–1941). The first study involved 3D imaging of sub-resolution fluorescent microspheres to determine the microscope point spread function. Results of the resolution studies as well as point spread function quality (i.e. objective lens quality) from 140 different objective lenses will be presented. The second study of spectral accuracy looked at the reflection of the laser excitation lines into the spectral detection in order to determine the accuracy of these systems to report back the accurate laser emission wavelengths. Results will be presented from 42 different spectral confocal systems. Finally, samples with double orange beads (orange core and orange coating) were imaged spectrally and the imaging software was used to un-mix fluorescence signals from the two orange dyes. Results from 26 different confocal systems will be summarized. Time will be left to discuss possibilities for the next LMRG study.

  6. Q: How do Microscopes Work?

    Science.gov (United States)

    Zimov, Sarah

    2004-01-01

    Microscopes allow scientists to examine everyday objects in extraordinary ways. They provide high-resolution images that show objects in fine detail. This brief article describes the many types of microscopes and how they are used in different scientific venues.

  7. Neutron microscope with refractive wedge

    International Nuclear Information System (INIS)

    Masalovich, S.V.

    1990-01-01

    A possibility of applying a refractive element in a mirror-neutron microscope using ultracold neutrons to reduce neutron aberrations is considered. Application of a refractive element in a neutron microscope with horizontal optical axis is studied. A scheme of neutron microscope with a refractive wedge is presented, evaluation of quartz wedge parameters is made. It is stressed that application of refractive elements in neutron microscopes facilitates aberration reduction in neutron-optical systems

  8. The Light Microscopy Module: An On-Orbit Multi-User Microscope Facility

    Science.gov (United States)

    Motil, Susan M.; Snead, John H.

    2002-01-01

    The Light Microscopy Module (LMM) is planned as a remotely controllable on-orbit microscope subrack facility, allowing flexible scheduling and operation of fluids and biology experiments within the Fluids and Combustion Facility (FCF) Fluids Integrated Rack (FIR) on the International Space Station (ISS). The LMM will be the first integrated payload with the FIR to conduct four fluid physics experiments. A description of the LMM diagnostic capabilities, including video microscopy, interferometry, laser tweezers, confocal, and spectrophotometry, will be provided.

  9. Atomic Force Microscope

    Energy Technology Data Exchange (ETDEWEB)

    Day, R.D.; Russell, P.E.

    1988-12-01

    The Atomic Force Microscope (AFM) is a recently developed instrument that has achieved atomic resolution imaging of both conducting and non- conducting surfaces. Because the AFM is in the early stages of development, and because of the difficulty of building the instrument, it is currently in use in fewer than ten laboratories worldwide. It promises to be a valuable tool for obtaining information about engineering surfaces and aiding the .study of precision fabrication processes. This paper gives an overview of AFM technology and presents plans to build an instrument designed to look at engineering surfaces.

  10. Solid state optical microscope

    Science.gov (United States)

    Young, Ian T.

    1983-01-01

    A solid state optical microscope wherein wide-field and high-resolution images of an object are produced at a rapid rate by utilizing conventional optics with a charge-coupled photodiode array. A galvanometer scanning mirror, for scanning in one of two orthogonal directions is provided, while the charge-coupled photodiode array scans in the other orthogonal direction. Illumination light from the object is incident upon the photodiodes, creating packets of electrons (signals) which are representative of the illuminated object. The signals are then processed, stored in a memory, and finally displayed as a video signal.

  11. Photography with a Microscope

    Science.gov (United States)

    Rost, Fred; Oldfield, Ron

    2000-03-01

    This beautifully illustrated book describes the methods used to record images viewed through a microscope. The text describes the principles and practices of photomicrography, and is written for all who take photomicrographs, beginners and/or experienced practitioners. The authors describe techniques that may be applied to many disciplines for teaching, research, archives, or pleasure. The book includes chapters on standard photography, modern digital techniques, methods for improving contrast, and a short chapter on drawing. In addition to its value as a work of reference, the authors' clear, didactic style makes this book suitable as a textbook for courses in photomicrography and/or elementary light microscopy.

  12. Confocal microscopy as an early relapse marker for acanthamoeba keratitis.

    Science.gov (United States)

    Daas, Loay; Viestenz, Arne; Schnabel, Philipp Albert; Fries, Fabian N; Hager, Tobias; SzentmÁry, Nora; Seitz, Berthold

    2018-01-01

    Acanthameoba keratitis is a serious ophthalmological condition with a potentially vision-threatening prognosis. Early diagnosis and recognition of relapse, and the detection of persistent Acanthamoeba cysts, are essential for informing the prognosis and managing the condition. We suggest the use of in vivo confocal microscopy not only to identify the early signs of relapse after keratoplasty in patients with Acanthamoeba keratitis, but also as an additional follow-up tool after antimicrobial crosslinking. This study shows that in vivo confocal microscopy is, in experienced hands, a quick and reliable diagnostic tool. Clin. Anat. 31:60-63, 2018. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  13. 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.

  14. DIY: "Do Imaging Yourself" - Conventional microscopes as powerful tools for in vivo investigation.

    Science.gov (United States)

    Antunes, Maísa Mota; Carvalho, Érika de; Menezes, Gustavo Batista

    2018-01-01

    Intravital imaging has been increasingly employed in cell biology studies and it is becoming one of the most powerful tools for in vivo investigation. Although some protocols can be extremely complex, most intravital imaging procedures can be performed using basic surgery and animal maintenance techniques. More importantly, regular confocal microscopes - the same that are used for imaging immunofluorescence slides - can also acquire high quality intravital images and movies after minor adaptations. Here we propose minimal adaptations in stock microscopes that allow major improvements in different fields of scientific investigation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Clinical management and microscopic characterisation of fatique-induced failure of a dental implant. Case report

    Directory of Open Access Journals (Sweden)

    De Frenza G

    2006-06-01

    Full Text Available Abstract Background Osseointegrated endosseous implants are widely used for the rehabilitation of completely and partially edentulous patients, being the final prosthodontic treatment more predictable and the failures extremely infrequent. A case of fracture of an endosseous dental implant, replacing the maxillary first molar, occurring in a middle-age woman, 5 years after placement is reported. Materials and methods The difficult management of this rare complication of implant dentistry together with the following rehabilitation is described. Additionally, the authors performed an accurate analysis of the removed fractured implant both by the stereomicroscope and by the confocal laser scanning microscope. Results and discussion The fractured impant showed the typical signs of a fatigue-induced fracture in the coronal portion of the implant together with numerous micro-fractures in the apical one. Three dimensional imaging performed by confocal laser scanning microscope led easily to a diagnosis of "fatigue fracture" of the implant. The biomechanical mechanism of implant fractures when overstress of the implant components due to bending overload is discussed. Conclusion When a fatigue-induced fracture of an dental implant occurs in presence of bending overload, the whole implant suffers a deformation that is confirmed by the alterations (micro-fractures of the implant observable also in the osseointegrated portion that is easily appraisable by the use of stereomicroscope and confocal laser scanning microscope without preparation of the sample.

  16. Nitrogen implantation with a scanning electron microscope.

    Science.gov (United States)

    Becker, S; Raatz, N; Jankuhn, St; John, R; Meijer, J

    2018-01-08

    Established techniques for ion implantation rely on technically advanced and costly machines like particle accelerators that only few research groups possess. We report here about a new and surprisingly simple ion implantation method that is based upon a widespread laboratory instrument: The scanning electron microscope. We show that it can be utilized to ionize atoms and molecules from the restgas by collisions with electrons of the beam and subsequently accelerate and implant them into an insulating sample by the effect of a potential building up at the sample surface. Our method is demonstrated by the implantation of nitrogen ions into diamond and their subsequent conversion to nitrogen vacancy centres which can be easily measured by fluorescence confocal microscopy. To provide evidence that the observed centres are truly generated in the way we describe, we supplied a 98% isotopically enriched 15 N gas to the chamber, whose natural abundance is very low. By employing the method of optically detected magnetic resonance, we were thus able to verify that the investigated centres are actually created from the 15 N isotopes. We also show that this method is compatible with lithography techniques using e-beam resist, as demonstrated by the implantation of lines using PMMA.

  17. Novel Multiwavelength Microscopic Scanner for Mouse Imaging

    Directory of Open Access Journals (Sweden)

    Herlen Alencar

    2005-11-01

    Full Text Available Real-time in vivo imaging of molecular targets at (subcellular resolution is essential in better understanding complex biology. Confocal microscopy and multiphoton microscopy have been used in the past to achieve this goal, but their true capabilities have often been limited by bulky optics and difficult experimental set-ups requiring exteriorized organs. We describe here the development and validation of a unique nearinfrared laser scanning microscope system that uses novel optics with a millimeter footprint. Optimized for use in the far red and near-infrared ranges, the system allows an imaging depth that extends up to 500 Mm from a 1.3-mm-diameter stick objective, which is up to 2 cm in length. We show exceptionally high spatial, temporal, and multiwavelength resolutions of the system and show that it can be applied to virtually any internal organ through a keyhole surgical access. We demonstrate that, when combined with novel far red imaging probes, it is possible to image the cellular details of many organs and disease processes. The new optics, coupled with the use of near-infrared probes, should prove immensely valuable for in vivo cancer imaging.

  18. Electron microscope phase enhancement

    Science.gov (United States)

    Jin, Jian; Glaeser, Robert M.

    2010-06-15

    A microfabricated electron phase shift element is used for modifying the phase characteristics of an electron beam passing though its center aperture, while not affecting the more divergent portion of an incident beam to selectively provide a ninety-degree phase shift to the unscattered beam in the back focal plan of the objective lens, in order to realize Zernike-type, in-focus phase contrast in an electron microscope. One application of the element is to increase the contrast of an electron microscope for viewing weakly scattering samples while in focus. Typical weakly scattering samples include biological samples such as macromolecules, or perhaps cells. Preliminary experimental images demonstrate that these devices do apply a ninety degree phase shift as expected. Electrostatic calculations have been used to determine that fringing fields in the region of the scattered electron beams will cause a negligible phase shift as long as the ratio of electrode length to the transverse feature-size aperture is about 5:1. Calculations are underway to determine the feasibility of aspect smaller aspect ratios of about 3:1 and about 2:1.

  19. Forensic Scanning Electron Microscope

    Science.gov (United States)

    Keeley, R. H.

    1983-03-01

    The scanning electron microscope equipped with an x-ray spectrometer is a versatile instrument which has many uses in the investigation of crime and preparation of scientific evidence for the courts. Major applications include microscopy and analysis of very small fragments of paint, glass and other materials which may link an individual with a scene of crime, identification of firearms residues and examination of questioned documents. Although simultaneous observation and chemical analysis of the sample is the most important feature of the instrument, other modes of operation such as cathodoluminescence spectrometry, backscattered electron imaging and direct x-ray excitation are also exploited. Marks on two bullets or cartridge cases can be compared directly by sequential scanning with a single beam or electronic linkage of two instruments. Particles of primer residue deposited on the skin and clothing when a gun is fired can be collected on adhesive tape and identified by their morphology and elemental composition. It is also possible to differentiate between the primer residues of different types of ammunition. Bullets may be identified from the small fragments left behind as they pass through the body tissues. In the examination of questioned documents the scanning electron microscope is used to establish the order in which two intersecting ink lines were written and to detect traces of chemical markers added to the security inks on official documents.

  20. Microinjection techniques for studying centrosome function in Drosophila melanogaster syncytial embryos.

    Science.gov (United States)

    Conduit, Paul T; Hayward, Daniel; Wakefield, James G

    2015-01-01

    Microinjection is a powerful technique that can be used to study protein function. Early Drosophila embryos are particularly amenable to microinjection due to their large size and their single cell status. Here, we report methods to microinject these embryos with various reagents to study the function of proteins at centrosomes and centrosome function more generally. Although precise details vary between laboratories, many aspects of the process are conserved. We describe the process from setting up a fly cage to imaging the injected embryos on a spinning disk confocal microscope and use specific examples to highlight the potency of this technique. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Anisotropic contrast optical microscope.

    Science.gov (United States)

    Peev, D; Hofmann, T; Kananizadeh, N; Beeram, S; Rodriguez, E; Wimer, S; Rodenhausen, K B; Herzinger, C M; Kasputis, T; Pfaunmiller, E; Nguyen, A; Korlacki, R; Pannier, A; Li, Y; Schubert, E; Hage, D; Schubert, M

    2016-11-01

    An optical microscope is described that reveals contrast in the Mueller matrix images of a thin, transparent, or semi-transparent specimen located within an anisotropic object plane (anisotropic filter). The specimen changes the anisotropy of the filter and thereby produces contrast within the Mueller matrix images. Here we use an anisotropic filter composed of a semi-transparent, nanostructured thin film with sub-wavelength thickness placed within the object plane. The sample is illuminated as in common optical microscopy but the light is modulated in its polarization using combinations of linear polarizers and phase plate (compensator) to control and analyze the state of polarization. Direct generalized ellipsometry data analysis approaches permit extraction of fundamental Mueller matrix object plane images dispensing with the need of Fourier expansion methods. Generalized ellipsometry model approaches are used for quantitative image analyses. These images are obtained from sets of multiple images obtained under various polarizer, analyzer, and compensator settings. Up to 16 independent Mueller matrix images can be obtained, while our current setup is limited to 11 images normalized by the unpolarized intensity. We demonstrate the anisotropic contrast optical microscope by measuring lithographically defined micro-patterned anisotropic filters, and we quantify the adsorption of an organic self-assembled monolayer film onto the anisotropic filter. Comparison with an isotropic glass slide demonstrates the image enhancement obtained by our method over microscopy without the use of an anisotropic filter. In our current instrument, we estimate the limit of detection for organic volumetric mass within the object plane of ≈49 fg within ≈7 × 7 μm 2 object surface area. Compared to a quartz crystal microbalance with dissipation instrumentation, where contemporary limits require a total load of ≈500 pg for detection, the instrumentation demonstrated here improves

  2. Axial Confocal Tomography of Capillary-Contained Colloidal Structures

    NARCIS (Netherlands)

    Liber, Shir R.; Indech, Ganit; Wee, Van Der Ernest B.; Butenko, Alexander V.; Kodger, Thomas E.; Lu, Peter J.; Schofield, Andrew B.; Weitz, David A.; Blaaderen, Van Alfons; Sloutskin, Eli

    2017-01-01

    Confocal microscopy is widely used for three-dimensional (3D) sample reconstructions. Arguably, the most significant challenge in such reconstructions is posed by the resolution along the optical axis being significantly lower than in the lateral directions. In addition, the imaging rate is lower

  3. Improvement in volume estimation from confocal sections after image deconvolution

    Czech Academy of Sciences Publication Activity Database

    Difato, Francesco; Mazzone, F.; Scaglione, S.; Fato, M.; Beltrame, F.; Kubínová, Lucie; Janáček, Jiří; Ramoino, P.; Vicidomini, G.; Diaspro, A.

    2004-01-01

    Roč. 64, č. 2 (2004), s. 151-155 ISSN 1059-910X Institutional research plan: CEZ:AV0Z5011922 Keywords : confocal microscopy * image deconvolution * point spread function Subject RIV: EA - Cell Biology Impact factor: 2.609, year: 2004

  4. Adaptive optics in digital micromirror based confocal microscopy

    NARCIS (Netherlands)

    Pozzi, P.; Wilding, D.; Soloviev, O.A.; Vdovine, Gleb; Verhaegen, M.H.G.; Bifano, Thomas G.; Kubby, Joel; Gigan, Sylvain

    2016-01-01

    This proceeding reports early results in the development of a new technique for adaptive optics in confocal microscopy. The term adaptive optics refers to the branch of optics in which an active element in the optical system is used to correct inhomogeneities in the media through which light

  5. Microscopic entropy and nonlocality

    International Nuclear Information System (INIS)

    Karpov, E.; Ordonets, G.; Petroskij, T.; Prigozhin, I.

    2003-01-01

    We have obtained a microscopic expression for entropy in terms of H function based on nonunitary Λ transformation which leads from the time evolution as a unitary group to a Markovian dynamics and unifies the reversible and irreversible aspects of quantum mechanics. This requires a new representation outside the Hilbert space. In terms of H, we show the entropy production and the entropy flow during the emission and absorption of radiation by an atom. Analyzing the time inversion experiment, we emphasize the importance of pre- and postcollisional correlations, which break the symmetry between incoming and outgoing waves. We consider the angle dependence of the H function in a three-dimensional situation. A model including virtual transitions is discussed in a subsequent paper

  6. Embryos, microscopes, and society.

    Science.gov (United States)

    Maienschein, Jane

    2016-06-01

    Embryos have different meanings for different people and in different contexts. Seen under the microscope, the biological embryo starts out as one cell and then becomes a bunch of cells. Gradually these divide and differentiate to make up the embryo, which in humans becomes a fetus at eight weeks, and then eventually a baby. At least, that happens in those cases that carry through normally and successfully. Yet a popular public perception imagines the embryo as already a little person in the very earliest stages of development, as if it were predictably to become an adult. In actuality, cells can combine, pull apart, and recombine in a variety of ways and still produce embryos, whereas most embryos never develop into adults at all. Biological embryos and popular imaginations of embryos diverge. This paper looks at some of the historical reasons for and social implications of that divergence. Copyright © 2016 The Author. Published by Elsevier Ltd.. All rights reserved.

  7. Imaging arrangement and microscope

    Science.gov (United States)

    Pertsinidis, Alexandros; Chu, Steven

    2015-12-15

    An embodiment of the present invention is an imaging arrangement that includes imaging optics, a fiducial light source, and a control system. In operation, the imaging optics separate light into first and second tight by wavelength and project the first and second light onto first and second areas within first and second detector regions, respectively. The imaging optics separate fiducial light from the fiducial light source into first and second fiducial light and project the first and second fiducial light onto third and fourth areas within the first and second detector regions, respectively. The control system adjusts alignment of the imaging optics so that the first and second fiducial light projected onto the first and second detector regions maintain relatively constant positions within the first and second detector regions, respectively. Another embodiment of the present invention is a microscope that includes the imaging arrangement.

  8. Microscopic Theory of Transconductivity

    Directory of Open Access Journals (Sweden)

    A. P. Jauho

    1998-01-01

    Full Text Available Measurements of momentum transfer between two closely spaced mesoscopic electronic systems, which couple via Coulomb interaction but where tunneling is inhibited, have proven to be a fruitful method of extracting information about interactions in mesoscopic systems. We report a fully microscopic theory for transconductivity σ12, or, equivalently, momentum transfer rate between the system constituents. Our main formal result expresses the transconductivity in terms of two fluctuation diagrams, which are topologically related, but not equivalent to, the Azlamazov-Larkin and Maki-Thompson diagrams known for superconductivity. In the present paper the magnetic field dependence of σ12 is discussed, and we find that σ12(B is strongly enhanced over its zero field value, and it displays strong features, which can be understood in terms of a competition between density-of-states and screening effects.

  9. Femtosecond photoelectron point projection microscope

    International Nuclear Information System (INIS)

    Quinonez, Erik; Handali, Jonathan; Barwick, Brett

    2013-01-01

    By utilizing a nanometer ultrafast electron source in a point projection microscope we demonstrate that images of nanoparticles with spatial resolutions of the order of 100 nanometers can be obtained. The duration of the emission process of the photoemitted electrons used to make images is shown to be of the order of 100 fs using an autocorrelation technique. The compact geometry of this photoelectron point projection microscope does not preclude its use as a simple ultrafast electron microscope, and we use simple analytic models to estimate temporal resolutions that can be expected when using it as a pump-probe ultrafast electron microscope. These models show a significant increase in temporal resolution when comparing to ultrafast electron microscopes based on conventional designs. We also model the microscopes spectroscopic abilities to capture ultrafast phenomena such as the photon induced near field effect

  10. Three-dimensional graphic reconstruction of the insect exoskeleton through confocal imaging of endogenous fluorescence.

    Science.gov (United States)

    Zill, S; Frazier, S F; Neff, D; Quimby, L; Carney, M; DiCaprio, R; Thuma, J; Norton, M

    2000-03-15

    The exoskeleton of the cockroach leg was imaged via confocal microscopy to generate digital graphic reconstructions of its three-dimensional structure. The cuticle is autofluorescent and can be visualized without staining, but is maximally imaged in aldehyde-fixed preparations viewed under krypton-argon laser illumination (yellow green (568 nm) excitation, commonly used in confocal microscopes). Images of the entire trochanteral segment of the leg were constructed as montages from optical sections taken as overlapping series that were coincident in the z-axis. Reconstructions of the exoskeleton from these images showed that strain sensing mechanoreceptors are located in association with buttresses and thickenings that form a consistent internal architecture in both juvenile and adult animals. Accuracy of reconstructions was gauged by embedding specimens in Spurr's resin and histologically sectioning them perpendicular to the optical plane of section (z-axis). Comparison of plastic sections with two-dimensional images generated by "resectioning" the software model showed that reconstructed exoskeleton had a high level of accuracy. Imaging of older and larger animals was limited by the sclerotization and increased thickness of the cuticle. Surface extraction algorithms were used to generate vector graphic files in CAD format for export to software used in engineering and design. Among other potential uses, these models have been studied by Finite Element Analysis to examine the distribution of mechanical strains in the exoskeleton that occur during posture and locomotion. The advantages and limitations of the techniques are discussed. These methods may be used in studying the exoskeleton and the anatomy of cuticular mechanoreceptors of other arthropods to similar advantage. Copyright 2000 Wiley-Liss, Inc.

  11. Scanning Electron Microscope Analysis System

    Data.gov (United States)

    Federal Laboratory Consortium — This facility provides the capability to examine surfaces microscopically with high resolution (5 nanometers), perform micro chemical analyses of these surfaces, and...

  12. Photon scanning tunneling microscope in combination with a force microscope

    NARCIS (Netherlands)

    Moers, M.H.P.; Moers, M.H.P.; Tack, R.G.; van Hulst, N.F.; Bölger, B.; Bölger, B.

    1994-01-01

    The simultaneous operation of a photon scanning tunneling microscope with an atomic force microscope is presented. The use of standard atomic force silicon nitride cantilevers as near-field optical probes offers the possibility to combine the two methods. Vertical forces and torsion are detected

  13. 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.

  14. 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.

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

    Science.gov (United States)

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

    2016-01-01

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

  16. The Latest in Handheld Microscopes

    Science.gov (United States)

    Wighting, Mervyn J.; Lucking, Robert A.; Christmann, Edwin P.

    2004-01-01

    Around 1590, Zacharias Jansenn of Holland invented the microscope. Jansenn, an eyeglass maker by trade, experimented with lenses and discovered that things appeared closer with combinations of lenses. Over the past 400 years, several refinements to microscopes have occurred, making it possible to magnify objects between 200 and 1,500 times their…

  17. Midfield microscope : Exploring the extraordinary

    NARCIS (Netherlands)

    Docter, M.W.

    2008-01-01

    In this thesis the development of the midfield microscope is presented. This is a microscope in which the extraordinary transmission (EOT) through sub-wavelength hole-arrays is applied. Before trying to combine microscopy and EOT, we look at them separately. In chapter 1 an overview is given of the

  18. Automated identification of epidermal keratinocytes in reflectance confocal microscopy

    Science.gov (United States)

    Gareau, Dan

    2011-03-01

    Keratinocytes in skin epidermis, which have bright cytoplasmic contrast and dark nuclear contrast in reflectance confocal microscopy (RCM), were modeled with a simple error function reflectance profile: erf( ). Forty-two example keratinocytes were identified as a training set which characterized the nuclear size a = 8.6+/-2.8 μm and reflectance gradient b = 3.6+/-2.1 μm at the nuclear/cytoplasmic boundary. These mean a and b parameters were used to create a rotationally symmetric erf( ) mask that approximated the mean keratinocyte image. A computer vision algorithm used an erf( ) mask to scan RCM images, identifying the coordinates of keratinocytes. Applying the mask to the confocal data identified the positions of keratinocytes in the epidermis. This simple model may be used to noninvasively evaluate keratinocyte populations as a quantitative morphometric diagnostic in skin cancer detection and evaluation of dermatological cosmetics.

  19. Confocal scanning laser ophthalmoscopy in glaucoma diagnosis and management.

    Science.gov (United States)

    Alexandrescu, C; Dascalu, A M; Panca, A; Sescioreanu, A; Mitulescu, C; Ciuluvica, R; Voinea, L; Celea, C

    2010-01-01

    The early diagnosis and detection of progression are two key-elements in the actual management of glaucoma. The current opinion in clinical practice is to quantify the structural damage for a better follow-up of the patient and the standardization of the results. The present review is a concise survey of literature covering the period of 1990-2010, documenting the evidence-based role of confocal scanning laser ophthalmoscopy in glaucoma diagnosis and management.

  20. In vivo intraoral reflectance confocal microscopy of an amalgam tattoo

    OpenAIRE

    Yélamos, Oriol; Cordova, Miguel; Peterson, Gary; Pulitzer, Melissa P.; Singh, Bhuvanesh; Rajadhyaksha, Milind; DeFazio, Jennifer L.

    2017-01-01

    The majority of oral pigmentations are benign lesions such as nevi, melanotic macules, melanoacanthomas or amalgam tattoos. Conversely, mucosal melanomas are rare but often lethal; therefore, excluding oral melanomas in this setting is crucial. Reflectance confocal microscopy is a non-invasive, in vivo imaging system with cellular resolution that has been used to distinguish benign from malignant pigmented lesions in the skin, and more recently in the mucosa. However, lesions located posterio...

  1. Line-scanning confocal microendoscope for nuclear morphometry imaging

    Science.gov (United States)

    Tang, Yubo; Carns, Jennifer; Richards-Kortum, Rebecca R.

    2017-11-01

    Fiber-optic endomicroscopy is a minimally invasive method to image cellular morphology in vivo. Using a coherent fiber bundle as an image relay, it allows additional imaging optics to be placed at the distal end of the fiber outside the body. In this research, we use this approach to demonstrate a compact, low-cost line-scanning confocal fluorescence microendoscope that can be constructed for pathological conditions.

  2. Adaptive optics in digital micromirror based confocal microscopy

    OpenAIRE

    Pozzi, P.; Wilding, D.; Soloviev, O.A.; Vdovine, Gleb; Verhaegen, M.H.G.; Bifano, Thomas G.; Kubby, Joel; Gigan, Sylvain

    2016-01-01

    This proceeding reports early results in the development of a new technique for adaptive optics in confocal microscopy. The term adaptive optics refers to the branch of optics in which an active element in the optical system is used to correct inhomogeneities in the media through which light propagates. In its most classical form, mostly used in astronomical imaging, adaptive optics is achieved through a closed loop in which the actuators of a deformable mirror are driven by a wavefront senso...

  3. The N-salicylidene aniline mesogen: Microscopic and macroscopic properties

    International Nuclear Information System (INIS)

    Nesrullazade, A.

    2004-01-01

    The vast majority of compounds exhibiting Iiquid crystalline phases may be regarded as having a rigid molecular central group with one or two flexible terminal alkyl or alkyloxy chains. The N-saIicyIidene anilines are very interesting and important materials both from fundamental and application points of view. These materials are on the one hand the ligands used to obtain metal containing complexes and on the other hand they are materials having the thermotropic mesomorphism. In this work we present investigations of microscopic and macroscopic properties of the 4-(Octyloxy)-N-(4-hexylphenyl)-2-hydrobenzaIimine (8SA) compound which was synthesized by our group. The 8SA compound shows the smectic C and nematic mesophases. These mesophases are enantiotropic and display specific confocal and schlieren textures, respectively. Thermotropic and thermodynamical properties of the straight and reverse phase transitions between smectic C and nematic mesophases and between nematic mesophase and isotropic liquid have been investigated

  4. Light Microscopy Module: International Space Station Premier Automated Microscope

    Science.gov (United States)

    Sicker, Ronald J.; Foster, William M.; Motil, Brian J.; Meyer, William V.; Chiaramonte, Francis P.; Abbott-Hearn, Amber; Atherton, Arthur; Beltram, Alexander; Bodzioney, Christopher; Brinkman, John; hide

    2016-01-01

    The Light Microscopy Module (LMM) was launched to the International Space Station (ISS) in 2009 and began hardware operations in 2010. It continues to support Physical and Biological scientific research on ISS. During 2016, if all goes as planned, three experiments will be completed: [1] Advanced Colloids Experiments with Heated base-2 (ACE-H2) and [2] Advanced Colloids Experiments with Temperature control (ACE-T1). Preliminary results, along with an overview of present and future LMM capabilities will be presented; this includes details on the planned data imaging processing and storage system, along with the confocal upgrade to the core microscope. [1] a consortium of universities from the State of Kentucky working through the Experimental Program to Stimulate Competitive Research (EPSCoR): Stuart Williams, Gerold Willing, Hemali Rathnayake, et al. and [2] from Chungnam National University, Daejeon, S. Korea: Chang-Soo Lee, et al.

  5. Setting up a simple light sheet microscope for in toto imaging of C. elegans development.

    Science.gov (United States)

    Chardès, Claire; Mélénec, Pauline; Bertrand, Vincent; Lenne, Pierre-François

    2014-05-05

    Fast and low phototoxic imaging techniques are pre-requisite to study the development of organisms in toto. Light sheet based microscopy reduces photo-bleaching and phototoxic effects compared to confocal microscopy, while providing 3D images with subcellular resolution. Here we present the setup of a light sheet based microscope, which is composed of an upright microscope and a small set of opto-mechanical elements for the generation of the light sheet. The protocol describes how to build, align the microscope and characterize the light sheet. In addition, it details how to implement the method for in toto imaging of C. elegans embryos using a simple observation chamber. The method allows the capture of 3D two-colors time-lapse movies over few hours of development. This should ease the tracking of cell shape, cell divisions and tagged proteins over long periods of time.

  6. Imaging the Microscopic Structure of Shear Thinning and Thickening Colloidal Suspensions

    KAUST Repository

    Cheng, X.

    2011-09-01

    The viscosity of colloidal suspensions varies with shear rate, an important effect encountered in many natural and industrial processes. Although this non-Newtonian behavior is believed to arise from the arrangement of suspended particles and their mutual interactions, microscopic particle dynamics are difficult to measure. By combining fast confocal microscopy with simultaneous force measurements, we systematically investigate a suspension\\'s structure as it transitions through regimes of different flow signatures. Our measurements of the microscopic single-particle dynamics show that shear thinning results from the decreased relative contribution of entropic forces and that shear thickening arises from particle clustering induced by hydrodynamic lubrication forces. This combination of techniques illustrates an approach that complements current methods for determining the microscopic origins of non-Newtonian flow behavior in complex fluids.

  7. The head-mounted microscope.

    Science.gov (United States)

    Chen, Ting; Dailey, Seth H; Naze, Sawyer A; Jiang, Jack J

    2012-04-01

    Microsurgical equipment has greatly advanced since the inception of the microscope into the operating room. These advancements have allowed for superior surgical precision and better post-operative results. This study focuses on the use of the Leica HM500 head-mounted microscope for the operating phonosurgeon. The head-mounted microscope has an optical zoom from 2× to 9× and provides a working distance from 300 mm to 700 mm. The headpiece, with its articulated eyepieces, adjusts easily to head shape and circumference, and offers a focus function, which is either automatic or manually controlled. We performed five microlaryngoscopic operations utilizing the head-mounted microscope with successful results. By creating a more ergonomically favorable operating posture, a surgeon may be able to obtain greater precision and success in phonomicrosurgery. Phonomicrosurgery requires the precise manipulation of long-handled cantilevered instruments through the narrow bore of a laryngoscope. The head-mounted microscope shortens the working distance compared with a stand microscope, thereby increasing arm stability, which may improve surgical precision. Also, the head-mounted design permits flexibility in head position, enabling operator comfort, and delaying musculoskeletal fatigue. A head-mounted microscope decreases the working distance and provides better ergonomics in laryngoscopic microsurgery. These advances provide the potential to promote precision in phonomicrosurgery. Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.

  8. Extrusion and intrusion evolution in cyclically strained cast superalloy Inconel 738LC using confocal laser scanning microscope and AFM

    Czech Academy of Sciences Publication Activity Database

    Obrtlík, Karel; Juliš, M.; Man, Jiří; Podrábský, T.; Polák, Jaroslav

    2010-01-01

    Roč. 241, - (2010), Art. No. 012054 ISSN 1742-6588. [ICSMA-15 (15th International Conference on the Strength of Materials). Dresden, 16.08.2009-21.08.2009] R&D Projects: GA AV ČR 1QS200410502; GA ČR GA106/07/1507 Institutional research plan: CEZ:AV0Z20410507 Keywords : fatigue * persistent slip marking * Inconel 738LC Subject RIV: JL - Materials Fatigue, Friction Mechanics http://iopscience.iop.org/1742-6596/240/1/012054

  9. Mobile microscope complex GIB-1

    International Nuclear Information System (INIS)

    Belyakov, A.V.; Gorbachev, A.N.

    2002-01-01

    To study microstructure in operating pipelines of power units a mobile microscope system is developed and successfully used. The system includes a portable microscope, a monitor, power supply and a portable computer. The monitor is used for surveying images from a video camera mounted on the microscope. The magnification on visual examination constitutes x 100 and x 500. Diameters of pipelines examined should not be less than 130 mm. Surface preparation for microstructural studies includes routine mechanical rough grinding and polishing with subsequent etching [ru

  10. Live confocal imaging of intrashell cytoplasm and calcification process in a symbiotic large benthic foraminifer Amphisorus kudakajimensis

    Science.gov (United States)

    Ohno, Yoshikazu; Fuita, Kazuhiko; Toyofuku, Takashi; Nakamura, Takashi

    2015-04-01

    Algal symbiont-bearing large benthic foraminifers are primary and carbonate producers as well as paleoenvironmental indicators in tropical and subtropical reef environments. Despite their importance, their cellular physiology is not well known. In this study, we have developed methods to observe in vivo images of a living symbiotic porcelaneous large foraminifer, Amphisorus kudakajimensis. Nikon A1 confocal laser scanning microscope with Calcein-AM as a fluorescent indicator was used to visualize morphology and streaming of cytoplasm in living A. kudakajimensis. The observation showed that in the marginal part of intrashell cytoplasm, cytoplasm gradually became highly branched and vacuolated. We also succeeded in observing elevated pH (pH 9.0) and lowered pH (pH 6.0) vesicles in reticulopodia-like cytoplasms, using a cell membrane-impermeable fluorescent pH indicator with low toxicity (HPTS). This study demonstrate the use of confocal microscopy in studying cytoplasmic dynamics and initial calcification process such as seawater endocytosis and alkalization of seawater vacuoles.

  11. Characterization of the stem anatomy of the Eocene fern Dennstaedtiopsis aerenchymata (Dennstaedtiaceae) by use of confocal laser scanning microscopy.

    Science.gov (United States)

    Shi, Chris S; Schopf, J William; Kudryavtsev, Anatoliy B

    2013-08-01

    Permineralization provides the most faithful known mode of three-dimensional preservation of the morphology and cellular anatomy of fossil plants. Standard optical microscopic documentation of such structures can provide only an approximation of their true three-dimensional form and is incapable of revealing fine-structural (<300 nm) details, deficiencies that can be addressed by the use of confocal laser scanning microscopy (CLSM). To demonstrate the usefulness of CLSM in such studies, we compare confocal laser scanning micrographs and optical photomicrographs of the permineralized tissues of rhizomes and petioles of the Eocene fern Dennstaedtiopsis aerenchymata preserved in cherts of the Clarno Formation of Oregon, USA, and the Allenby Formation (Princeton chert) of British Columbia, Canada. The laser-induced fluorescence detected by CLSM produces crisp high-resolution images of the three-dimensionally permineralized tissues of Dennstaedtiopsis aerenchymata. Tissues analyzed include the epidermis and epidermal hairs, cortex, aerenchyma, endodermis, vascular tissue, and pith-for each of which, CLSM yields results superior to those of standard optical microscopy. CLSM and previous Raman spectroscopic analyses of the same specimens provide evidence consistent with original biochemistry. Use of CLSM to characterize the morphology and cellular anatomy of permineralized fossil plants can provide accurate data in two and three dimensions at high spatial resolution, information that can be critically important to taxonomic, taphonomic, and developmental interpretations. Results presented here from this first detailed CLSM-based study of permineralized plant axes indicate that this nonintrusive, nondestructive technique should be widely applicable in paleobotany.

  12. 3D optical sectioning with a new hyperspectral confocal fluorescence imaging system.

    Energy Technology Data Exchange (ETDEWEB)

    Nieman, Linda T.; Sinclair, Michael B.; Davidson, George S.; Van Benthem, Mark Hilary; Haaland, David Michael; Timlin, Jerilyn Ann; Sasaki, Darryl Yoshio; Bachand, George David; Jones, Howland D. T.

    2007-02-01

    A novel hyperspectral fluorescence microscope for high-resolution 3D optical sectioning of cells and other structures has been designed, constructed, and used to investigate a number of different problems. We have significantly extended new multivariate curve resolution (MCR) data analysis methods to deconvolve the hyperspectral image data and to rapidly extract quantitative 3D concentration distribution maps of all emitting species. The imaging system has many advantages over current confocal imaging systems including simultaneous monitoring of numerous highly overlapped fluorophores, immunity to autofluorescence or impurity fluorescence, enhanced sensitivity, and dramatically improved accuracy, reliability, and dynamic range. Efficient data compression in the spectral dimension has allowed personal computers to perform quantitative analysis of hyperspectral images of large size without loss of image quality. We have also developed and tested software to perform analysis of time resolved hyperspectral images using trilinear multivariate analysis methods. The new imaging system is an enabling technology for numerous applications including (1) 3D composition mapping analysis of multicomponent processes occurring during host-pathogen interactions, (2) monitoring microfluidic processes, (3) imaging of molecular motors and (4) understanding photosynthetic processes in wild type and mutant Synechocystis cyanobacteria.

  13. Detection of wood cell wall porosity using small carbohydrate molecules and confocal fluorescence microscopy.

    Science.gov (United States)

    Donaldson, L A; Kroese, H W; Hill, S J; Franich, R A

    2015-09-01

    A novel approach to nanoscale detection of cell wall porosity using confocal fluorescence microscopy is described. Infiltration of cell walls with a range of nitrophenyl-substituted carbohydrates of different molecular weights was assessed by measuring changes in the intensity of lignin fluorescence, in response to the quenching effect of the 4-nitrophenyl group. The following carbohydrates were used in order of increasing molecular weight; 4-nitrophenyl β-D-glucopyrano-side (monosaccharide), 4-nitrophenyl β-D-lactopyranoside (disaccharide), 2-chloro-4-nitrophenyl β-D-maltotrioside (trisaccharide), and 4-nitrophenyl α-D-maltopentaoside (pentasaccharide). This technique was used to compare cell wall porosity in wood which had been dewatered to 40% moisture content using supercritical CO2, where cell walls remain fully hydrated, with kiln dried wood equilibrated to 12% moisture content. Infiltration of cell walls as measured by fluorescence quenching, was found to decrease with increasing molecular weight, with the pentasaccharide being significantly excluded compared to the monosaccharide. Porosity experiments were performed on blocks and sections to assess differences in cell wall accessibility. Dewatered and kiln dried wood infiltrated as blocks showed similar results, but greater infiltration was achieved by using sections, indicating that not all pores were easily accessible by infiltration from the lumen surface. In wood blocks infiltrated with 4-nitrophenyl α-D-maltopentaoside, quenching of the secondary wall was quite variable, especially in kiln dried wood, indicating limited connectivity of pores accessible from the lumen surface. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

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

    Directory of Open Access Journals (Sweden)

    KA. Al-Salihi

    2009-12-01

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

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

    Science.gov (United States)

    Martisek, Dalibor; Prochazkova, Jana

    2017-12-01

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

  16. Localization of extracellular matrix components in developing mouse salivary glands by confocal microscopy

    Science.gov (United States)

    Hardman, P.; Spooner, B. S.

    1992-01-01

    The importance of the extracellular matrix (ECM) in epithelial-mesenchymal interactions in developing organisms is well established. Proteoglycans and interstitial collagens are required for the growth, morphogenesis, and differentiation of epithelial organs and the distribution of these molecules has been described. However, much less is known about other ECM macromolecules in developing epithelial organs. We used confocal microscopy to examine the distribution of laminin, heparan sulfate (BM-1) proteoglycan, fibronectin, and collagen types I, IV, and V, in mouse embryonic salivary glands. Organ rudiments were isolated from gestational day 13 mouse embryos and cultured for 24, 48, or 72 hours. Whole mounts were stained by indirect immunofluorescence and then examined using a Zeiss Laser Scan Microscope. We found that each ECM component examined had a distinct distribution and that the distribution of some molecules varied with culture time. Laminin was mainly restricted to the basement membrane. BM-1 proteoglycan was concentrated in the basement membrane and also formed a fine network throughout the mesenchyme. Type IV collagen was mainly located in the basement membrane of the epithelium, but it was also present throughout the mesenchyme. Type V collagen was distributed throughout the mesenchyme at 24 hours, but at 48 hours was principally located in the basement membrane. Type I collagen was distributed throughout the mesenchyme at all culture times, and accumulated in the clefts and particularly at the epithelial-mesenchymal interface as time in culture increased. Fibronectin was observed throughout the mesenchyme at all times.

  17. Confocal Microscopy Monitoring of pH Dynamics in a 5-Species Dental Biofilm Model Under Flow Conditions

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Dige, Irene

    2017-01-01

    -species biofilm model of dental caries under static and flow conditions. Biofilms were grown in the absence of sucrose in ibidi µ-slides for 30 h or 120 h. Biofilms were then stained with the ratiometric probe C-SNARF-4 at pH 7.0, incubated with 0.4% glucose, and z-stacks were acquired in 9 microscopic......Measuring pH dynamics in dental biofilms is of utmost importance for a thorough understanding of the caries process. We have developed a confocal microscopy based methodology to monitor pH in biofilms at the microscale in real-time. Here, we measure pH developments in different layers of a 5...

  18. Microscopic examination of deteriorated concrete

    NARCIS (Netherlands)

    Nijland, T.G.; Larbi, J.A.

    2010-01-01

    Concrete petrography is the integrated microscopic and mesoscale (hand specimen size) investigation of hardened concrete, that can provide information on the composition of concrete, the original relationships between the concrete's various constituents, and any changes therein, whether as a result

  19. Center of excellence: Microlaser microscope

    Science.gov (United States)

    Webb, R. H.

    This Center-of-Excellence grant has two components: development of an imaging system based on microlaser arrays forms a central project among a group of laser diagnostic and therapeutic efforts primarily funded outside the grant. In these first 8 months we have set up the Microlaser Microscope using small microlaser arrays. We have emphasized the basics of microlaser handling and electronic addressing and the optics of the microscope. Details of electronics and optics given here will be used in the larger arrays which should be available soon. After a description of the central Microlaser Microscope project, we touch briefly on the other projects of the Center, which have been outstandingly fruitful this year. Publications are necessarily concerned with the smaller projects, since the Microlaser Microscope is in its early stages.

  20. In vivo skin characterization by confocal Raman microspectroscopy

    NARCIS (Netherlands)

    P.J. Caspers (Peter)

    2003-01-01

    markdownabstract__Abstract__ Various areas of skin research depend on detailed knowledge of the molecular composition of skin and molecular structure of skin constituents. On a microscopic scale the skin is a highly heterogeneous tissue. Molecular composition and structure vary

  1. On thermodynamic and microscopic reversibility

    International Nuclear Information System (INIS)

    Crooks, Gavin E

    2011-01-01

    The word 'reversible' has two (apparently) distinct applications in statistical thermodynamics. A thermodynamically reversible process indicates an experimental protocol for which the entropy change is zero, whereas the principle of microscopic reversibility asserts that the probability of any trajectory of a system through phase space equals that of the time reversed trajectory. However, these two terms are actually synonymous: a thermodynamically reversible process is microscopically reversible, and vice versa

  2. Characterization of a hazardous eyeliner (kohl) by confocal Raman microscopy

    International Nuclear Information System (INIS)

    Jallad, Karim N.; Hedderich, Hartmut G.

    2005-01-01

    A new method of analyzing kohl, a cosmetic eyeliner, using confocal Raman microscopy is reported. This technique offers an important alternative to conventional spectroscopic techniques that provide elemental/atomic composition. Raman spectra of three kohl samples have been measured between 150 and 3000 cm -1 at room temperature. The main component of two kohl samples was found to be lead(II) sulfide (PbS). Kohl is used as a traditional cosmetic and remedy in the Middle East, Far East, and Northern Africa. Since kohl products contain very high concentrations of lead, they constitute a risk for public health, particularly for children

  3. Volume visualization of biological tissue specimens using confocal microscopy

    Czech Academy of Sciences Publication Activity Database

    Čapek, Martin; Janáček, Jiří; Kubínová, Lucie; Smrčka, P.; Hána, K.

    2006-01-01

    Roč. 36, č. 2 (2006), s. 240-244 ISSN 0301-5491. [Biomedical Engineering Conference of Young Biomedical Engineers and Researchers /2./. Kladno, 19.07.2006-21.07.2006] R&D Projects: GA MŠk(CZ) LC06063; GA AV ČR(CZ) IAA100110502; GA AV ČR(CZ) IAA500200510; GA ČR(CZ) GA304/05/0153 Institutional research plan: CEZ:AV0Z50110509 Keywords : 3D reconstruction * confocal microscopy Subject RIV: JC - Computer Hardware ; Software

  4. 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

  5. Aerial wetting contact angle measurement using confocal microscopy

    OpenAIRE

    Chesna, Jacob W.; Wiedmaier, Bob F.; Wang, Jinlin; Samara, Ayman; Leach, Richard K.; Her, Tsing-Hua; Smith, Stuart T.

    2016-01-01

    A method is presented in which the wetting contact angle of a sessile drop is acquired aerially using confocal techniques to measure the radius and the height of a droplet deposited on a planar surface. The repeatability of this method is typically less than 0.25°, and often less than 0.1°, for droplet diameters less than 1 mm. To evaluate accuracy of this method, an instrument uncertainty budget is developed, which predicts a combined uncertainty of 0.91° for a 1 mm diameter water droplet wi...

  6. Enlightening the Pink: Use of Confocal Microscopy in Pink Lesions.

    Science.gov (United States)

    Gill, Melissa; González, Salvador

    2016-10-01

    Solitary pink lesions can pose a particular challenge to dermatologists because they may be almost or completely featureless clinically and dermoscopically, previously requiring biopsy to exclude malignancy. However, these lesions usually are not particularly challenging histopathologically. Thus, the incorporation of in vivo reflectance confocal microscopy into the clinical practice, which allows for noninvasive examination of the skin at the cellular level revealing features previously seen only on histopathology, is particularly useful for this subset of clinically difficult lesions. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Scanning Miniature Microscopes without Lenses

    Science.gov (United States)

    Wang, Yu

    2009-01-01

    The figure schematically depicts some alternative designs of proposed compact, lightweight optoelectronic microscopes that would contain no lenses and would generate magnified video images of specimens. Microscopes of this type were described previously in Miniature Microscope Without Lenses (NPO - 20218), NASA Tech Briefs, Vol. 22, No. 8 (August 1998), page 43 and Reflective Variants of Miniature Microscope Without Lenses (NPO 20610), NASA Tech Briefs, Vol. 26, No. 9 (September 1999), page 6a. To recapitulate: In the design and construction of a microscope of this type, the focusing optics of a conventional microscope are replaced by a combination of a microchannel filter and a charge-coupled-device (CCD) image detector. Elimination of focusing optics reduces the size and weight of the instrument and eliminates the need for the time-consuming focusing operation. The microscopes described in the cited prior articles contained two-dimensional CCDs registered with two-dimensional arrays of microchannels and, as such, were designed to produce full two-dimensional images, without need for scanning. The microscopes of the present proposal would contain one-dimensional (line image) CCDs registered with linear arrays of microchannels. In the operation of such a microscope, one would scan a specimen along a line perpendicular to the array axis (in other words, one would scan in pushbroom fashion). One could then synthesize a full two-dimensional image of the specimen from the line-image data acquired at one-pixel increments of position along the scan. In one of the proposed microscopes, a beam of unpolarized light for illuminating the specimen would enter from the side. This light would be reflected down onto the specimen by a nonpolarizing beam splitter attached to the microchannels at their lower ends. A portion of the light incident on the specimen would be reflected upward, through the beam splitter and along the microchannels, to form an image on the CCD. If the

  8. Video-mosaicking of in vivo reflectance confocal microscopy images for noninvasive examination of skin lesion (Conference Presentation)

    Science.gov (United States)

    Kose, Kivanc; Gou, Mengran; Yelamos, Oriol; Cordova, Miguel A.; Rossi, Anthony; Nehal, Kishwer S.; Camps, Octavia I.; Dy, Jennifer G.; Brooks, Dana H.; Rajadhyaksha, Milind

    2017-02-01

    In this report we describe a computer vision based pipeline to convert in-vivo reflectance confocal microscopy (RCM) videos collected with a handheld system into large field of view (FOV) mosaics. For many applications such as imaging of hard to access lesions, intraoperative assessment of MOHS margins, or delineation of lesion margins beyond clinical borders, raster scan based mosaicing techniques have clinically significant limitations. In such cases, clinicians often capture RCM videos by freely moving a handheld microscope over the area of interest, but the resulting videos lose large-scale spatial relationships. Videomosaicking is a standard computational imaging technique to register, and stitch together consecutive frames of videos into large FOV high resolution mosaics. However, mosaicing RCM videos collected in-vivo has unique challenges: (i) tissue may deform or warp due to physical contact with the microscope objective lens, (ii) discontinuities or "jumps" between consecutive images and motion blur artifacts may occur, due to manual operation of the microscope, and (iii) optical sectioning and resolution may vary between consecutive images due to scattering and aberrations induced by changes in imaging depth and tissue morphology. We addressed these challenges by adapting or developing new algorithmic methods for videomosaicking, specifically by modeling non-rigid deformations, followed by automatically detecting discontinuities (cut locations) and, finally, applying a data-driven image stitching approach that fully preserves resolution and tissue morphologic detail without imposing arbitrary pre-defined boundaries. We will present example mosaics obtained by clinical imaging of both melanoma and non-melanoma skin cancers. The ability to combine freehand mosaicing for handheld microscopes with preserved cellular resolution will have high impact application in diverse clinical settings, including low-resource healthcare systems.

  9. Small fiber neuropathy in women with fibromyalgia. An in vivo assessment using corneal confocal bio-microscopy.

    Science.gov (United States)

    Ramírez, Manuel; Martínez-Martínez, Laura-Aline; Hernández-Quintela, Everardo; Velazco-Casapía, Jorge; Vargas, Angélica; Martínez-Lavín, Manuel

    2015-10-01

    A consistent line of investigation suggests that fibromyalgia is a neuropathic pain syndrome. This outlook has been recently reinforced by several controlled studies that describe decreased small nerve fiber density in skin biopsies of patients with fibromyalgia. The cornea receives the densest small fiber innervation of the body. Corneal confocal bio-microscopy is a new noninvasive method to evaluate small nerve fiber morphology. Our objective was to assess corneal small nerve fiber morphology in patients with fibromyalgia, and to associate corneal nerve microscopic features with neuropathic pain descriptors and other fibromyalgia symptoms. We studied 17 female patients with fibromyalgia and 17 age-matched healthy control subjects. All the participants completed different questionnaires regarding the symptoms of fibromyalgia, including a neuropathic pain survey. A central corneal thickness scan was obtained with a confocal microscope. Nerve measurements were made by a single ophthalmologist without knowledge of the clinical diagnosis. Stromal nerve thickness was defined as the mean value between the widest and the narrowest portion of each analyzed stromal nerve. Corneal sub-basal plexus nerve density was also assessed. Patients with fibromyalgia had stromal nerve thickness of 5.0 ± 1.0 µm (mean ± standard deviation) significantly different from that of control's values (6.1 ± 1.3) p = 0.01. Patients also had decreased sub-basal plexus nerve density per square millimeter (85 ± 29) vs. 107 ± 26 of controls p = 0.02. When controls and patients were grouped together, there was an association between stromal nerve slenderness and neuropathic pain descriptors (Fisher's exact test p = 0.007). Women suffering from fibromyalgia have thinner corneal stromal nerves and diminished sub-basal plexus nerve density when compared to healthy controls. Nerve scarcity is associated with neuropathic pain descriptors. Small fiber neuropathy may play a role in the pathogenesis of

  10. Microscopia confocal de la córnea en facoemulsificación Confocal microscopy of the cornea on phacoemulsification

    Directory of Open Access Journals (Sweden)

    Juan Raúl Hernández Silva

    2011-12-01

    Full Text Available Objetivo: Determinar los cambios estructurales de la córnea en la cirugía de catarata por facoemulsificación sin complicaciones. Métodos: Se realizó un estudio prospectivo de pacientes operados de catarata por facoemulsificación coaxial por la técnica de pre chop sin complicaciones. A estos se les realizó microscopia confocal de la córnea con el CONFOSCAN 4 (Nidek Technologies con el objetivo de 40x y adaptador Z-Ring. Se realizó el estudio en el preoperatorio y en el posoperatorio (a las 24 horas, después de una semana, de un mes y a los tres meses. Resultados: Se demostraron cambios estructurales en la córnea como células epiteliales con núcleos hiperreflectivos alargadas en ocasiones y áreas de hiperreflectividad anómala a las 24 horas del posoperatorio. Persistieron queratocitos activados y la disminución de la hiperreflectividad de la matriz extracelular que desapareció al mes. Conclusiones: Aunque por biomicroscopia no se observen alteraciones corneales en el posoperatorio de la cirugía de catarata por facoemulsificación, sí se pueden demostrar por microscopia confocal de la córnea. Estas variaciones no influyen en la recuperación visual óptima de los pacientes.Objective: To determine the structural changes in the cornea in the cataract surgery using phacoemulsification without complications. Methods: A prospective study of patients operated on from cataract using the coaxial phacoemulsification (Pre Chop technique without complications was carried out. These patients also underwent confocal microscopy of the cornea with Confoscan4 (Nidek Technologies with 40x target and Z - Ring adapter. The study was performed in the preoperative period and postoperative period for 24 hours, one week, one month and three months after surgery. Results: Structural changes were observed in the cornea such as epithelial cells with hypereflectivity nucleus, occasionally elongated, , areas of anomalous hypereflectivity 24 hours after

  11. Embryonic Heart Morphogenesis from Confocal Microscopy Imaging and Automatic Segmentation

    Directory of Open Access Journals (Sweden)

    Hongda Mao

    2013-01-01

    Full Text Available Embryonic heart morphogenesis (EHM is a complex and dynamic process where the heart transforms from a single tube into a four-chambered pump. This process is of great biological and clinical interest but is still poorly understood for two main reasons. On the one hand, the existing imaging modalities for investigating EHM suffered from either limited penetration depth or limited spatial resolution. On the other hand, current works typically adopted manual segmentation, which was tedious, subjective, and time consuming considering the complexity of developing heart geometry and the large size of images. In this paper, we propose to utilize confocal microscopy imaging with tissue optical immersion clearing technique to image the heart at different stages of development for EHM study. The imaging method is able to produce high spatial resolution images and achieve large penetration depth at the same time. Furthermore, we propose a novel convex active contour model for automatic image segmentation. The model has the ability to deal with intensity fall-off in depth which is characterized by confocal microscopy images. We acquired the images of embryonic quail hearts from day 6 to day 14 of incubation for EHM study. The experimental results were promising and provided us with an insight view of early heart growth pattern and also paved the road for data-driven heart growth modeling.

  12. Resolution enhancement in confocal microscopy using Bessel-Gauss beams.

    Science.gov (United States)

    Thibon, Louis; Lorenzo, Louis E; Piché, Michel; De Koninck, Yves

    2017-02-06

    Laser scanning microscopy is limited in lateral resolution by the diffraction of light. We show that we can obtain twenty percent improvement in the resolution of confocal microscopy using Bessel-Gauss beams with the right pinhole size compared to conventional Gaussian beam based confocal microscopy. Advantages of this strategy include simplicity of installation and use, linear polarization compatibility, possibility to combine it with other resolution enhancement and superresolution strategies. We demonstrate the resolution enhancement capabilities of Bessel-Gauss beams both theoretically and experimentally on nano-spheres and biological tissue samples without any residual artifacts coming from the Bessel-Gauss beam side lobes with a resolution of 0.39λ. We also show that the resolution enhancement of Bessel-Gauss beams yields a better statistical colocalization analysis with fewer false positive results than when using Gaussian beams. We have also used Bessel-Gauss beams of different orders to further improve the resolution by combining them in SLAM microscopy (Switching LAser Modes : Dehez, Optics Express, 2013) achieving a resolution of 0.2λ.

  13. Axiomatic electrodynamics and microscopic mechanics

    International Nuclear Information System (INIS)

    Yussouff, M.

    1981-04-01

    A new approach to theoretical physics, along with the basic formulation of a new MICROSCOPIC MECHANICS for the motion of small charged particles is described in this set of lecture notes. Starting with the classical (Newtonian) mechanics and classical fields, the important but well known properties of Classical Electromagnetic field are discussed up to section 4. The next nection describes the usual radiation damping theory and its difficulties. It is argued that the usual treatment of radiation damping is not valid for small space and time intervals and the true description of motion requires a new type of mechanics - the MICROSCOPIC MECHANICS: Section 6 and 7 are devoted to showing that not only the new microscopic mechanics goes over to Newtonian mechanics in the proper limit, but also it is closely connected with Quantum Mechanics. All the known results of the Schroedinger theory can be reproduced by microscopic mechanics which also gives a clear physical picture. It removes Einstein's famous objections against Quantum Theory and provides a clear distinction between classical and Quantum behavior. Seven Axioms (three on Classical Mechanics, two for Maxwell's theory, one for Relativity and a new Axiom on Radiation damping) are shown to combine Classical Mechanics, Maxwellian Electrodynamics, Relativity and Schroedinger's Quantum Theory within a single theoretical framework under Microscopic Mechanics which awaits further development at the present time. (orig.)

  14. Confocal absorption spectral imaging of MoS2: optical transitions depending on the atomic thickness of intrinsic and chemically doped MoS2.

    Science.gov (United States)

    Dhakal, Krishna P; Duong, Dinh Loc; Lee, Jubok; Nam, Honggi; Kim, Minsu; Kan, Min; Lee, Young Hee; Kim, Jeongyong

    2014-11-07

    We performed a nanoscale confocal absorption spectral imaging to obtain the full absorption spectra (over the range 1.5-3.2 eV) within regions having different numbers of layers and studied the variation of optical transition depending on the atomic thickness of the MoS2 film. Three distinct absorption bands corresponding to A and B excitons and a high-energy background (BG) peak at 2.84 eV displayed a gradual redshift as the MoS2 film thickness increased from the monolayer, to the bilayer, to the bulk MoS2 and this shift was attributed to the reduction of the gap energy in the Brillouin zone at the K-point as the atomic thickness increased. We also performed n-type chemical doping of MoS2 films using reduced benzyl viologen (BV) and the confocal absorption spectra modified by the doping showed a strong dependence on the atomic thickness: A and B exciton peaks were greatly quenched in the monolayer MoS2 while much less effect was shown in larger thickness and the BG peak either showed very small quenching for 1 L MoS2 or remained constant for larger thicknesses. Our results indicate that confocal absorption spectral imaging can provide comprehensive information on optical transitions of microscopic size intrinsic and doped two-dimensional layered materials.

  15. Normative values for corneal nerve morphology assessed using corneal confocal microscopy: a multinational normative data set.

    Science.gov (United States)

    Tavakoli, Mitra; Ferdousi, Maryam; Petropoulos, Ioannis N; Morris, Julie; Pritchard, Nicola; Zhivov, Andrey; Ziegler, Dan; Pacaud, Danièle; Romanchuk, Kenneth; Perkins, Bruce A; Lovblom, Leif E; Bril, Vera; Singleton, J Robinson; Smith, Gordon; Boulton, Andrew J M; Efron, Nathan; Malik, Rayaz A

    2015-05-01

    Corneal confocal microscopy is a novel diagnostic technique for the detection of nerve damage and repair in a range of peripheral neuropathies, in particular diabetic neuropathy. Normative reference values are required to enable clinical translation and wider use of this technique. We have therefore undertaken a multicenter collaboration to provide worldwide age-adjusted normative values of corneal nerve fiber parameters. A total of 1,965 corneal nerve images from 343 healthy volunteers were pooled from six clinical academic centers. All subjects underwent examination with the Heidelberg Retina Tomograph corneal confocal microscope. Images of the central corneal subbasal nerve plexus were acquired by each center using a standard protocol and analyzed by three trained examiners using manual tracing and semiautomated software (CCMetrics). Age trends were established using simple linear regression, and normative corneal nerve fiber density (CNFD), corneal nerve fiber branch density (CNBD), corneal nerve fiber length (CNFL), and corneal nerve fiber tortuosity (CNFT) reference values were calculated using quantile regression analysis. There was a significant linear age-dependent decrease in CNFD (-0.164 no./mm(2) per year for men, P < 0.01, and -0.161 no./mm(2) per year for women, P < 0.01). There was no change with age in CNBD (0.192 no./mm(2) per year for men, P = 0.26, and -0.050 no./mm(2) per year for women, P = 0.78). CNFL decreased in men (-0.045 mm/mm(2) per year, P = 0.07) and women (-0.060 mm/mm(2) per year, P = 0.02). CNFT increased with age in men (0.044 per year, P < 0.01) and women (0.046 per year, P < 0.01). Height, weight, and BMI did not influence the 5th percentile normative values for any corneal nerve parameter. This study provides robust worldwide normative reference values for corneal nerve parameters to be used in research and clinical practice in the study of diabetic and other peripheral neuropathies. © 2015 by the American Diabetes Association

  16. Application of confocal X-ray fluorescence micro-spectroscopy to the investigation of paint layers.

    Science.gov (United States)

    Sun, Tianxi; Liu, Zhiguo; Wang, Guangfu; Ma, Yongzhong; Peng, Song; Sun, Weiyuan; Li, Fangzuo; Sun, Xuepeng; Ding, Xunliang

    2014-12-01

    A confocal micro X-ray fluorescence (MXRF) spectrometer based on polycapillary X-ray optics was used for the identification of paint layers. The performance of the confocal MXRF was studied. Multilayered paint fragments of a car were analyzed nondestructively to demonstrate that this confocal MXRF instrument could be used in the discrimination of the various layers in multilayer paint systems. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Microscope and method of use

    Science.gov (United States)

    Bongianni, Wayne L.

    1984-01-01

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

  18. Atomic force microscope featuring an integrated optical microscope

    NARCIS (Netherlands)

    Putman, C.A.J.; Putman, Constant A.J.; de Grooth, B.G.; van Hulst, N.F.; Greve, Jan

    1992-01-01

    The atomic force microscope (AFM) is used to image the surface of both conductors and nonconductors. Biological specimens constitute a large group of nonconductors. A disadvantage of most AFM's is the fact that relatively large areas of the sample surface have to be scanned to pinpoint a biological

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

    Science.gov (United States)

    Attota, Ravi Kiran; Park, Haesung

    2017-06-15

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

  20. Optical sectioning microscopes with no moving parts using a micro-stripe array light emitting diode.

    Science.gov (United States)

    Poher, V; Zhang, H X; Kennedy, G T; Griffin, C; Oddos, S; Gu, E; Elson, D S; Girkin, M; French, P M W; Dawson, M D; Neil, M A

    2007-09-03

    We describe an optical sectioning microscopy system with no moving parts based on a micro-structured stripe-array light emitting diode (LED). By projecting arbitrary line or grid patterns onto the object, we are able to implement a variety of optical sectioning microscopy techniques such as grid-projection structured illumination and line scanning confocal microscopy, switching from one imaging technique to another without modifying the microscope setup. The micro-structured LED and driver are detailed and depth discrimination capabilities are measured and calculated.

  1. Infrared up-conversion microscope

    DEFF Research Database (Denmark)

    2014-01-01

    There is presented an up-conversion infrared microscope (110) arranged for imaging an associated object (130), wherein the up-conversion infrared microscope (110) comprises a non-linear crystal (120) arranged for up-conversion of infrared electromagnetic radiation, and wherein an objective optical...... component (100) has an entrance pupil with a first diameter D1, and an optical component system which is arranged for forming an external image (136) of the back-focal plane (132) of the objective optical component (100), which has a diameter (given by the diameter of a circle enclosing all optical paths...

  2. 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

  3. 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

  4. Research on improving performance to metallographic microscopes

    Directory of Open Access Journals (Sweden)

    Teodor Socaciu

    2011-12-01

    Full Text Available Precision optical components from an old optical microscope can be improved and emphasized with a dedicated digital microscope camera. This is an affordable way to obtain a high performance metallographic or biological microscope, with minimum of spending. This paper study those ways and adapts a camera to existing microscopes for researchers use, Optimizing visualization by projecting the image and improving the microscope use by different options of capture and image processing.

  5. Curriculum Guidelines for Microscopic Anatomy.

    Science.gov (United States)

    Journal of Dental Education, 1993

    1993-01-01

    The American Association of Dental Schools' guidelines for curricula in microscopic anatomy offer an overview of the histology curriculum, note primary educational goals, outline specific content for general and oral histology, suggest prerequisites, and make recommendations for sequencing. Appropriate faculty and facilities are also suggested.…

  6. Microscopic cross sections: An utopia?

    Energy Technology Data Exchange (ETDEWEB)

    Hilaire, S. [CEA Bruyeres-le-Chatel, DIF 91 (France); Koning, A.J. [Nuclear Research and Consultancy Group, PO Box 25, 1755 ZG Petten (Netherlands); Goriely, S. [Institut d' Astronomie et d' Astrophysique, Universite Libre de Bruxelles, Campus de la Plaine, CP 226, 1050 Brussels (Belgium)

    2010-07-01

    The increasing need for cross sections far from the valley of stability poses a challenge for nuclear reaction models. So far, predictions of cross sections have relied on more or less phenomenological approaches, depending on parameters adjusted to available experimental data or deduced from systematical relations. While such predictions are expected to be reliable for nuclei not too far from the experimentally known regions, it is clearly preferable to use more fundamental approaches, based on sound physical bases, when dealing with very exotic nuclei. Thanks to the high computer power available today, all major ingredients required to model a nuclear reaction can now be (and have been) microscopically (or semi-microscopically) determined starting from the information provided by a nucleon-nucleon effective interaction. We have implemented all these microscopic ingredients in the TALYS nuclear reaction code, and we are now almost able to perform fully microscopic cross section calculations. The quality of these ingredients and the impact of using them instead of the usually adopted phenomenological parameters will be discussed. (authors)

  7. Microscopic cross sections: An utopia?

    International Nuclear Information System (INIS)

    Hilaire, S.; Koning, A.J.; Goriely, S.

    2010-01-01

    The increasing need for cross sections far from the valley of stability poses a challenge for nuclear reaction models. So far, predictions of cross sections have relied on more or less phenomenological approaches, depending on parameters adjusted to available experimental data or deduced from systematical relations.While such predictions are expected to be reliable for nuclei not too far from the experimentally known regions, it is clearly preferable to use more fundamental approaches, based on sound physical bases, when dealing with very exotic nuclei. Thanks to the high computer power available today, all major ingredients required to model a nuclear reaction can now be (and have been) microscopically (or semi-microscopically) determined starting from the information provided by a nucleon-nucleon effective interaction. We have implemented all these microscopic ingredients in the TALYS nuclear reaction code, and we are now almost able to perform fully microscopic cross section calculations. The quality of these ingredients and the impact of using them instead of the usually adopted phenomenological parameters will be discussed. (authors)

  8. Microscope sterility during spine surgery.

    Science.gov (United States)

    Bible, Jesse E; O'Neill, Kevin R; Crosby, Colin G; Schoenecker, Jonathan G; McGirt, Matthew J; Devin, Clinton J

    2012-04-01

    Prospective study. Assess the contamination rates of sterile microscope drapes after spine surgery. The use of the operating microscope has become more prevalent in certain spine procedures, providing superior magnification, visualization, and illumination of the operative field. However, it may represent an additional source of bacterial contamination and increase the risk of developing a postoperative infection. This study included 25 surgical spine cases performed by a single spine surgeon that required the use of the operative microscope. Sterile culture swabs were used to obtain samples from 7 defined locations on the microscope drape after its use during the operation. The undraped technician's console was sampled in each case as a positive control, and an additional 25 microscope drapes were swabbed immediately after they were applied to the microscope to obtain negative controls. Swab samples were assessed for bacterial growth on 5% sheep blood Columbia agar plates using a semiquantitative technique. No growth was observed on any of the 25 negative control drapes. In contrast, 100% of preoperative and 96% of postoperative positive controls demonstrated obvious contamination. In the postoperative group, all 7 sites of evaluation were found to be contaminated with rates of 12% to 44%. Four of the 7 evaluated locations were found to have significant contamination rates compared with negative controls, including the shafts of the optic eyepieces on the main surgeon side (24%, P = 0.022), "forehead" portion on both the main surgeon (24%, P = 0.022) and assistant sides (28%, P = 0.010), and "overhead" portion of the drape (44%, P = 0.0002). Bacterial contamination of the operative microscope was found to be significant after spine surgery. Contamination was more common around the optic eyepieces, likely due to inadvertent touching of unsterile portions. Similarly, all regions above the eyepieces also have a propensity for contamination because of unknown contact

  9. 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.

  10. In vivo intraoral reflectance confocal microscopy of an amalgam tattoo.

    Science.gov (United States)

    Yélamos, Oriol; Cordova, Miguel; Peterson, Gary; Pulitzer, Melissa P; Singh, Bhuvanesh; Rajadhyaksha, Milind; DeFazio, Jennifer L

    2017-10-01

    The majority of oral pigmentations are benign lesions such as nevi, melanotic macules, melanoacanthomas or amalgam tattoos. Conversely, mucosal melanomas are rare but often lethal; therefore, excluding oral melanomas in this setting is crucial. Reflectance confocal microscopy is a non-invasive, in vivo imaging system with cellular resolution that has been used to distinguish benign from malignant pigmented lesions in the skin, and more recently in the mucosa. However, lesions located posteriorly in the oral cavity are difficult to assess visually and difficult to biopsy due to their location. Herein we present a patient with previous multiple melanomas presenting with an oral amalgam tattoo in the buccal mucosa, which was imaged using an intraoral telescopic probe attached to a commercially available handheld RCM. In this case report we describe this novel probe, the first RCM description of an amalgam tattoo and we discuss its differences with the findings described in oral melanomas.

  11. Materials and corrosion characterization using the confocal resonator

    Energy Technology Data Exchange (ETDEWEB)

    Tigges, C.P.; Sorensen, N.R.; Hietala, V.M.; Plut, T.A. [and others

    1997-05-01

    Improved characterization and process control is important to many Sandia and DOE programs related to manufacturing. Many processes/structures are currently under-characterized including thin film growth, corrosion and semiconductor structures, such as implant profiles. A sensitive tool is required that is able to provide lateral and vertical imaging of the electromagnetic properties of a sample. The confocal resonator is able to characterize the surface and near-surface impedance of materials. This device may be applied to a broad range of applications including in situ evaluation of thin film processes, physical defect detection/characterization, the characterization of semiconductor devices and corrosion studies. In all of these cases, the technology should work as a real-time process diagnostic or as a feedback mechanism regarding the quality of a manufacturing process. This report summarizes the development and exploration of several diagnostic applications.

  12. Characterization of Developing Cotton Fibers by Confocal Raman Microscopy

    Directory of Open Access Journals (Sweden)

    Luis Cabrales

    2014-10-01

    Full Text Available Cellulose deposition in developing cotton fibers has been studied previously with analytical techniques, such as Fourier transform infrared spectroscopy (FTIR, High-performance liquid chromatography (HPLC and Thermogravimetric analysis (TGA. Recent technological developments in instrumentation have made Raman microscopy emerge as an extraordinary analytical tool in biological and plant research. The advantage of using confocal Raman microscopy (CRM resides in the lateral spatial resolution and in the fact that Raman spectroscopy provides not only chemical composition information, but also structural information. Cross-sections of cotton fibers harvested at different developmental stages were studied with CRM. The Raman bands assigned to cellulose were analyzed. The results of this study indicate that CRM can be used as a tool to study cellulose deposition in cotton fibers and could provide useful information on cellulose deposition during cotton fiber development.

  13. Confocal mapping of myelin figures with micro-Raman spectroscopy

    Science.gov (United States)

    Huang, Jung-Ren; Cheng, Yu-Che; Huang, Hung Ji; Chiang, Hai-Pang

    2018-01-01

    We employ confocal micro-Raman spectroscopy (CMRS) with submicron spatial resolution to study the myelin structures (cylindrical lamellae) composed of nested surfactant C12E3 or lipid DMPC bilayers. The CMRS mapping indicates that for a straight C12E3 myelin, the surfactant concentration increases with the myelin width and is higher in the center region than in the peripheral region. For a curved C12E3 myelin, the convex side has a higher surfactant concentration than the corresponding concave side. The spectrum of DMPC myelins undergoes a qualitative change as the temperature increases above 60 °C, suggesting that the surfactant molecules may be damaged. Our work demonstrates the utility of CMRS in bio-soft material research.

  14. The Reliability and Reproducibility of Corneal Confocal Microscopy in Children.

    Science.gov (United States)

    Pacaud, Danièle; Romanchuk, Kenneth G; Tavakoli, Mitra; Gougeon, Claire; Virtanen, Heidi; Ferdousi, Maryam; Nettel-Aguirre, Alberto; Mah, Jean K; Malik, Rayaz A

    2015-08-01

    To assess the image and patient level interrater agreement and repeatability within 1 month for corneal nerve fiber length (CNFL) measured using in vivo corneal confocal microscopy (IVCCM) in children. Seventy-one subjects (mean [SD] age 14.3 [2.6] years, range 8-18 years; 44 with type 1 diabetes and 27 controls; 36 males and 35 females) were included. 547 images (∼6 images per subject) were analyzed manually by two independent and masked observers. One-month repeat visit images were analyzed by a single masked observer in 21 patients. Automated image analysis was then performed using a specialized computerized software (ACCMetrics). For CNFL, the ICC (95% CI) were 0.94 (0.93-0.95) for image-level, 0.86 (0.78-0.91) for patient-level, and 0.88 (0.72-0.95) for the 1-month repeat assessment, and the Bland-Altman plots showed minimal bias between observers. Although there was excellent agreement between manual and automated analysis according to an ICC 0.89 (0.82-0.93), the Bland-Altman plot showed a consistent bias with manual measurements providing higher readings. In vivo corneal confocal microscopy image analysis shows good reproducibility with excellent intraindividual and interindividual variability in pediatric subjects. Since the image-level reproducibility is stronger than the patient-level reproducibility, refinement of the method for image selection will likely further increase the robustness of this novel, rapid, and noninvasive approach to detect early neuropathy in children with diabetes. Further studies on the use of IVCCM to identify early subclinical neuropathy in children are indicated.

  15. Endoscopic probe optics for spectrally encoded confocal microscopy.

    Science.gov (United States)

    Kang, Dongkyun; Carruth, Robert W; Kim, Minkyu; Schlachter, Simon C; Shishkov, Milen; Woods, Kevin; Tabatabaei, Nima; Wu, Tao; Tearney, Guillermo J

    2013-01-01

    Spectrally encoded confocal microscopy (SECM) is a form of reflectance confocal microscopy that can achieve high imaging speeds using relatively simple probe optics. Previously, the feasibility of conducting large-area SECM imaging of the esophagus in bench top setups has been demonstrated. Challenges remain, however, in translating SECM into a clinically-useable device; the tissue imaging performance should be improved, and the probe size needs to be significantly reduced so that it can fit into luminal organs of interest. In this paper, we report the development of new SECM endoscopic probe optics that addresses these challenges. A custom water-immersion aspheric singlet (NA = 0.5) was developed and used as the objective lens. The water-immersion condition was used to reduce the spherical aberrations and specular reflection from the tissue surface, which enables cellular imaging of the tissue deep below the surface. A custom collimation lens and a small-size grating were used along with the custom aspheric singlet to reduce the probe size. A dual-clad fiber was used to provide both the single- and multi- mode detection modes. The SECM probe optics was made to be 5.85 mm in diameter and 30 mm in length, which is small enough for safe and comfortable endoscopic imaging of the gastrointestinal tract. The lateral resolution was 1.8 and 2.3 µm for the single- and multi- mode detection modes, respectively, and the axial resolution 11 and 17 µm. SECM images of the swine esophageal tissue demonstrated the capability of this device to enable the visualization of characteristic cellular structural features, including basal cell nuclei and papillae, down to the imaging depth of 260 µm. These results suggest that the new SECM endoscopic probe optics will be useful for imaging large areas of the esophagus at the cellular scale in vivo.

  16. A versatile optical microscope for time-dependent single-molecule and single-particle spectroscopy.

    Science.gov (United States)

    Li, Hao; Yang, Haw

    2018-03-28

    This work reports the design and implementation of a multi-function optical microscope for time-dependent spectroscopy on single molecules and single nanoparticles. It integrates the now-routine single-object measurements into one standalone platform so that no reconfiguration is needed when switching between different types of sample or spectroscopy modes. The illumination modes include evanescent field through total internal reflection, dark-field illumination, and epi-excitation onto a diffraction-limited spot suitable for confocal detection. The detection modes include spectrally resolved line imaging, wide-field imaging with dual-color capability, and two-color single-element photon-counting detection. The switch between different spectroscopy and data acquisition modes is fully automated and executed through computer programming. The capability of this microscope is demonstrated through selected proof-of-principle experiments.

  17. A versatile optical microscope for time-dependent single-molecule and single-particle spectroscopy

    Science.gov (United States)

    Li, Hao; Yang, Haw

    2018-03-01

    This work reports the design and implementation of a multi-function optical microscope for time-dependent spectroscopy on single molecules and single nanoparticles. It integrates the now-routine single-object measurements into one standalone platform so that no reconfiguration is needed when switching between different types of sample or spectroscopy modes. The illumination modes include evanescent field through total internal reflection, dark-field illumination, and epi-excitation onto a diffraction-limited spot suitable for confocal detection. The detection modes include spectrally resolved line imaging, wide-field imaging with dual-color capability, and two-color single-element photon-counting detection. The switch between different spectroscopy and data acquisition modes is fully automated and executed through computer programming. The capability of this microscope is demonstrated through selected proof-of-principle experiments.

  18. Theory of Linear and Nonlinear Gain in a Gyroamplifier using a Confocal Waveguide.

    Science.gov (United States)

    Soane, Alexander V; Shapiro, Michael A; Stephens, Jacob C; Temkin, Richard J

    2017-09-01

    The linear and nonlinear theory of a gyroamplifier using a confocal waveguide is presented. A quasi-optical approach to describing the modes of a confocal waveguide is derived. Both the equations of motion and the mode excitation equation are derived in detail. The confocal waveguide circuit has the advantage of reducing mode competition but the lack of azimuthal symmetry presents challenges in calculating the gain. In the linear regime, the gain calculated using the exact form factor for the confocal waveguide agrees with an azimuthally averaged form factor. A beamlet code including velocity spread effects has been written to calculate the linear and nonlinear (saturated) gain. It has been successfully benchmarked against the MAGY code for azimuthally symmetric cases. For the confocal waveguide, the beamlet code shows that the saturated gain is reduced when compared with results obtained using an azimuthally averaged form factor. The beamlet code derived here extends the capabilities of nonlinear gyroamplifier theory to configurations that lack azimuthal symmetry.

  19. Improving spatial resolution of confocal Raman microscopy by super-resolution image restoration.

    Science.gov (United States)

    Cui, Han; Zhao, Weiqian; Wang, Yun; Fan, Ying; Qiu, Lirong; Zhu, Ke

    2016-05-16

    A new super-resolution image restoration confocal Raman microscopy method (SRIR-RAMAN) is proposed for improving the spatial resolution of confocal Raman microscopy. This method can recover the lost high spatial frequency of the confocal Raman microscopy by using Poisson-MAP super-resolution imaging restoration, thereby improving the spatial resolution of confocal Raman microscopy and realizing its super-resolution imaging. Simulation analyses and experimental results indicate that the spatial resolution of SRIR-RAMAN can be improved by 65% to achieve 200 nm with the same confocal Raman microscopy system. This method can provide a new tool for high spatial resolution micro-probe structure detection in physical chemistry, materials science, biomedical science and other areas.

  20. 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

  1. 21 CFR 884.6190 - Assisted reproductive microscopes and microscope accessories.

    Science.gov (United States)

    2010-04-01

    ... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES OBSTETRICAL AND GYNECOLOGICAL DEVICES Assisted Reproduction.... Assisted reproduction microscopes and microscope accessories (excluding microscope stage warmers, which are classified under assisted reproduction accessories) are optical instruments used to enlarge images of gametes...

  2. Differential magnetic force microscope imaging.

    Science.gov (United States)

    Wang, Ying; Wang, Zuobin; Liu, Jinyun; Hou, Liwei

    2015-01-01

    This paper presents a method for differential magnetic force microscope imaging based on a two-pass scanning procedure to extract differential magnetic forces and eliminate or significantly reduce background forces with reversed tip magnetization. In the work, the difference of two scanned images with reversed tip magnetization was used to express the local magnetic forces. The magnetic sample was first scanned with a low lift distance between the MFM tip and the sample surface, and the magnetization direction of the probe was then changed after the first scan to perform the second scan. The differential magnetic force image was obtained through the subtraction of the two images from the two scans. The theoretical and experimental results have shown that the proposed method for differential magnetic force microscope imaging is able to reduce the effect of background or environment interference forces, and offers an improved image contrast and signal to noise ratio (SNR). © Wiley Periodicals, Inc.

  3. Infrared up-conversion microscope

    DEFF Research Database (Denmark)

    2014-01-01

    There is presented an up-conversion infrared microscope (110) arranged for imaging an associated object (130), wherein the up-conversion infrared microscope (110) comprises a non-linear crystal (120) arranged for up-conversion of infrared electromagnetic radiation, and wherein an objective optical...... component (100) has an entrance pupil with a first diameter D1, and an optical component system which is arranged for forming an external image (136) of the back-focal plane (132) of the objective optical component (100), which has a diameter (given by the diameter of a circle enclosing all optical paths...... at the plane of the 10 external image) which is denominated D2 and wherein D1 is larger than a second diameter D2....

  4. Duties to Extraterrestrial Microscopic Organisms

    Science.gov (United States)

    Cockell, C. S.

    Formulating a normative axiology for the treatment of extraterrestrial microscopic organisms, should they ever be found, requires an extension of environmental ethics to beyond the Earth. Using an ethical framework for the treatment of terrestrial micro-organisms, this paper elaborates a similar ethic for the treatment of extraterrestrial microscopic organisms. An ethic of `teloempathy' allows for the moral considerability of any organism that has `interests', based on rudimentary qualities of conativism, and therefore allows for an identical treatment of all life, related or not related to life on Earth. Although, according to this ethic, individual extraterrestrial microscopic organisms have a good of their own and even `rights', at this level the ethic can only be theoretical, allowing for the inevitable destruction of many individual organisms during the course of human exploratory missions, similarly to the daily destruction of microbes by humans on Earth. A holistic teloempathy, an operative ethic, not only provides a framework for human exploration, but it also has important implications for planetary protection and proposals to implement planetary-scale atmospheric alterations on other bodies. Even prior to the discovery of extraterrestrial life, or the discovery of a complete absence of such life, this exercise yields important insights into the moral philosophy that guides our treatment of terrestrial micro-organisms.

  5. The nematode stoma: Homology of cell architecture with improved understanding by confocal microscopy of labeled cell boundaries.

    Science.gov (United States)

    Jay Burr, A H; Baldwin, James G

    2016-09-01

    Nematode stomas vary widely in the cuticular structures evolved for different feeding strategies, yet the arrangement of the epithelial cell classes that form these structures may be conserved. This article addresses several issues that have impeded the full acceptance of this hypothesis including controversies arising from the structure of the Caenorhabditis elegans stoma. We investigated fluorescent antibody labeling of cell boundaries in conjunction with confocal microscopy as an alternative to transmission electron microscopy (TEM), using MH27 to label apical junctions in C. elegans and two other species. Accurately spaced optical sections collected by the confocal microscope provide a three-dimensional array of pixels (voxels) that, using image-processing software, can be rotated and sectioned at accurately chosen thicknesses and locations. Ribbons of fluorescence clearly identify cell boundaries along the luminal cuticle in C. elegans and Zeldia punctata and less clearly in Bunonema sp. The patterns render cell classes and their relationships readily identifiable. In the C. elegans stoma they correct a misreading of serial TEMs that was not congruent with architecture in other nematodes-the row of marginal cells is now seen to be continuous as in other nematodes, rather than being interrupted by encircling pm1 cells. Also impeding understanding, the reference to certain cell classes as 'epithelial' and others as "muscle" in the C. elegans literature is at variance with muscle expression in most other taxa. For consistent comparison among species, we propose that these cell class descriptors based on function be replaced by topological terms. With these and other confusing concepts and terminology removed, the homology of the cellular architecture among taxa becomes obvious. We provide a corrected description of the cell architecture of the C. elegans stoma and examples of how it is modified in other taxa with different feeding strategies. J. Morphol. 277

  6. Development of HiLo Microscope and its use in In-Vivo Applications

    Science.gov (United States)

    Patel, Shreyas J.

    The functionality of achieving optical sectioning in biomedical research is invaluable as it allows for visualization of a biological sample at different depths while being free of background scattering. Most current microscopy techniques that offer optical sectioning, unfortunately, require complex instrumentation and thus are generally costly. HiLo microscopy, on the other hand, offers the same functionality and advantage at a relatively low cost. Hence, the work described in this thesis involves the design, build, and application of a HiLo microscope. More specifically, a standalone HiLo microscope was built in addition to implementing HiLo microscopy on a standard fluorescence microscope. In HiLo microscopy, optical sectioning is achieved by acquiring two different types of images per focal plane. One image is acquired under uniform illumination and the other is acquired under speckle illumination. These images are processed using an algorithm that extracts in-focus information and removes features and glare that occur as a result of background fluorescence. To show the benefits of the HiLo microscopy, several imaging experiments on various samples were performed under a HiLo microscope and compared against a traditional fluorescence microscope and a confocal microscope, which is considered the gold standard in optical imaging. In-vitro and ex-vivo imaging was performed on a set of pollen grains, and optically cleared mouse brain and heart slices. Each of these experiments showed great reduction in background scattering at different depths under HiLo microscopy. More importantly, HiLo imaging of optically cleared heart slice demonstrated emergence of different vasculature at different depths. Reduction of out-of-focus light increased the spatial resolution and allowed better visualization of capillary vessels. Furthermore, HiLo imaging was tested in an in-vivo model of a rodent dorsal window chamber model. When imaging the same sample under confocal microscope

  7. Confocal Raman microscopy and multivariate statistical analysis for determination of different penetration abilities of caffeine and propylene glycol applied simultaneously in a mixture on porcine skin ex vivo.

    Science.gov (United States)

    Mujica Ascencio, Saul; Choe, ChunSik; Meinke, Martina C; Müller, Rainer H; Maksimov, George V; Wigger-Alberti, Walter; Lademann, Juergen; Darvin, Maxim E

    2016-07-01

    Propylene glycol is one of the known substances added in cosmetic formulations as a penetration enhancer. Recently, nanocrystals have been employed also to increase the skin penetration of active components. Caffeine is a component with many applications and its penetration into the epidermis is controversially discussed in the literature. In the present study, the penetration ability of two components - caffeine nanocrystals and propylene glycol, applied topically on porcine ear skin in the form of a gel, was investigated ex vivo using two confocal Raman microscopes operated at different excitation wavelengths (785nm and 633nm). Several depth profiles were acquired in the fingerprint region and different spectral ranges, i.e., 526-600cm(-1) and 810-880cm(-1) were chosen for independent analysis of caffeine and propylene glycol penetration into the skin, respectively. Multivariate statistical methods such as principal component analysis (PCA) and linear discriminant analysis (LDA) combined with Student's t-test were employed to calculate the maximum penetration depths of each substance (caffeine and propylene glycol). The results show that propylene glycol penetrates significantly deeper than caffeine (20.7-22.0μm versus 12.3-13.0μm) without any penetration enhancement effect on caffeine. The results confirm that different substances, even if applied onto the skin as a mixture, can penetrate differently. The penetration depths of caffeine and propylene glycol obtained using two different confocal Raman microscopes are comparable showing that both types of microscopes are well suited for such investigations and that multivariate statistical PCA-LDA methods combined with Student's t-test are very useful for analyzing the penetration of different substances into the skin. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Literature survey on microscopic friction modeling

    NARCIS (Netherlands)

    Hol, J.

    2010-01-01

    To better understand contact and friction conditions, experimental and theoretical studies have been performed in order to take microscopic dependencies into account. Friction is developed on microscopic level by adhesion between contacting asperities, the ploughing effect between asperities and the

  9. A new method for depth profiling reconstruction in confocal microscopy

    Science.gov (United States)

    Esposito, Rosario; Scherillo, Giuseppe; Mensitieri, Giuseppe

    2018-05-01

    Confocal microscopy is commonly used to reconstruct depth profiles of chemical species in multicomponent systems and to image nuclear and cellular details in human tissues via image intensity measurements of optical sections. However, the performance of this technique is reduced by inherent effects related to wave diffraction phenomena, refractive index mismatch and finite beam spot size. All these effects distort the optical wave and cause an image to be captured of a small volume around the desired illuminated focal point within the specimen rather than an image of the focal point itself. The size of this small volume increases with depth, thus causing a further loss of resolution and distortion of the profile. Recently, we proposed a theoretical model that accounts for the above wave distortion and allows for a correct reconstruction of the depth profiles for homogeneous samples. In this paper, this theoretical approach has been adapted for describing the profiles measured from non-homogeneous distributions of emitters inside the investigated samples. The intensity image is built by summing the intensities collected from each of the emitters planes belonging to the illuminated volume, weighed by the emitters concentration. The true distribution of the emitters concentration is recovered by a new approach that implements this theoretical model in a numerical algorithm based on the Maximum Entropy Method. Comparisons with experimental data and numerical simulations show that this new approach is able to recover the real unknown concentration distribution from experimental profiles with an accuracy better than 3%.

  10. Spatial resolution of confocal XRF technique using capillary optics.

    Science.gov (United States)

    Dehlinger, Maël; Fauquet, Carole; Lavandier, Sebastien; Aumporn, Orawan; Jandard, Franck; Arkadiev, Vladimir; Bjeoumikhov, Aniouar; Tonneau, Didier

    2013-06-07

    XRF (X-ray fluorescence) is a powerful technique for elemental analysis with a high sensitivity. The resolution is presently limited by the size of the primary excitation X-ray beam. A test-bed for confocal-type XRF has been developed to estimate the ultimate lateral resolution which could be reached in chemical mapping using this technique. A polycapillary lens is used to tightly focus the primary X-ray beam of a low power rhodium X-ray source, while the fluorescence signal is collected by a SDD detector through a cylindrical monocapillary. This system was used to characterize the geometry of the fluorescent zone. Capillary radii ranging from 50 μm down to 5 μm were used to investigate the fluorescence signal maximum level This study allows to estimate the ultimate resolution which could be reached in-lab or on a synchrotron beamline. A new tool combining local XRF and scanning probe microscopy is finally proposed.

  11. Confocal microscopy for intracellular co-localization of proteins.

    Science.gov (United States)

    Miyashita, Toshiyuki

    2015-01-01

    Confocal laser scanning microscopy is the best method to visualize intracellular co-localization of proteins in intact cells. Because of the point scan/pinhole detection system, light contribution from the neighborhood of the scanning spot in the specimen can be eliminated, allowing high Z-axis resolution. Fluorescence detection by sensitive photomultiplier tubes allows the usage of filters with a narrow bandpath, resulting in minimal cross-talk (overlap) between two spectra. This is particularly important in demonstrating co-localization of proteins with multicolor labeling. Here, the methods outlining the detection of transiently expressed tagged proteins and the detection of endogenous proteins are described. Ideally, the intracellular co-localization of two endogenous proteins should be demonstrated. However, when antibodies raised against the protein of interest are unavailable for immunofluorescence or the available cell lines do not express the protein of interest sufficiently enough for immunofluorescence, an alternative method is to transfect cells with expression plasmids that encode tagged proteins and stain the cells with anti-tag antibodies. However, it should be noted that the tagging of proteins of interest or their overexpression could potentially alter the intracellular localization or the function of the target protein.

  12. A Student-Built Scanning Tunneling Microscope

    Science.gov (United States)

    Ekkens, Tom

    2015-01-01

    Many introductory and nanotechnology textbooks discuss the operation of various microscopes including atomic force (AFM), scanning tunneling (STM), and scanning electron microscopes (SEM). In a nanotechnology laboratory class, students frequently utilize microscopes to obtain data without a thought about the detailed operation of the tool itself.…

  13. Microscopic Analysis of Activated Sludge. Training Manual.

    Science.gov (United States)

    Office of Water Program Operations (EPA), Cincinnati, OH. National Training and Operational Technology Center.

    This training manual presents material on the use of a compound microscope to analyze microscope communities, present in wastewater treatment processes, for operational control. Course topics include: sampling techniques, sample handling, laboratory analysis, identification of organisms, data interpretation, and use of the compound microscope.…

  14. 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.

  15. 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.

  16. Comparative measurement of collagen bundle orientation by Fourier analysis and semiquantitative evaluation: reliability and agreement in Masson's trichrome, Picrosirius red and confocal microscopy techniques.

    Science.gov (United States)

    Marcos-Garcés, V; Harvat, M; Molina Aguilar, P; Ferrández Izquierdo, A; Ruiz-Saurí, A

    2017-08-01

    agreement with H&E-confocal (0.7 0.8), better with Fourier analysis than with semiquantitative evaluation (single and average). These results in nonpathological skin samples were also confirmed in a preliminary analysis in eight scleroderma skin samples. Our results show that Masson's trichrome and Picrosirius red are consistent with H&E-confocal for measuring collagen bundle orientation in histological samples and could thus be used indistinctly for this purpose. Fourier analysis is superior to average semiquantitative evaluation and should keep being used as the preferred method. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  17. A novel optical microscope for imaging large embryos and tissue volumes with sub-cellular resolution throughout.

    Science.gov (United States)

    McConnell, Gail; Trägårdh, Johanna; Amor, Rumelo; Dempster, John; Reid, Es; Amos, William Bradshaw

    2016-09-23

    Current optical microscope objectives of low magnification have low numerical aperture and therefore have too little depth resolution and discrimination to perform well in confocal and nonlinear microscopy. This is a serious limitation in important areas, including the phenotypic screening of human genes in transgenic mice by study of embryos undergoing advanced organogenesis. We have built an optical lens system for 3D imaging of objects up to 6 mm wide and 3 mm thick with depth resolution of only a few microns instead of the tens of microns currently attained, allowing sub-cellular detail to be resolved throughout the volume. We present this lens, called the Mesolens, with performance data and images from biological specimens including confocal images of whole fixed and intact fluorescently-stained 12.5-day old mouse embryos.

  18. Microscopic structure for light nuclei

    International Nuclear Information System (INIS)

    Sharma, V.K.

    1995-01-01

    The microscopic structure for light nuclei e.g. 4 He, 7 Li and 8 Be is considered in the frame work of the generator coordinate method (GCM). The physical interpretation of our GCM is also discussed. The GC amplitudes are used to calculate the various properties like charge and magnetic RMS radii, form factors, electromagnetic moments, astrophysical S-factor, Bremsstrahlung weighted cross sections, relative wavefunctions and vertex functions etc. All the calculated quantities agree well with the values determined experimentally. (author). 30 refs., 10 figs., 2 tabs

  19. Microscopic structure of liquid hydrogen

    CERN Document Server

    Zoppi, M

    2003-01-01

    Hydrogen makes the simplest molecular liquid. Nonetheless, due to several different reasons, measuring its microscopic structure has been one of the most challenging tasks in neutron diffraction experiments. The recent development of modern pulsed neutron sources triggered a renewed experimental interest which, in turn, led to new knowledge and also to a more effective use of the classic reactor-based experimental data. The contemporary development of quantum mechanical computer simulation techniques, and a critical comparison among the results of different experiments using steady and pulsed neutron sources, resulted in a quantitatively reliable solution of the problem. (topical review)

  20. Confocal X-ray fluorescence spectrometer for in-situ analyses of paintings

    Science.gov (United States)

    Trojek, Tomáš; Prokeš, Radek; Šefců, Radka; Bilavčíková, Hana; Čechák, Tomáš

    2017-08-01

    This paper describes the properties of the newly constructed device for confocal X-ray fluorescence analysis that was tested with a sample plate consisting of 19 combinations of two single pigment layers. The preparation of this experimental wooden board with layered systems was based on knowledge of the panel painting techniques of Bohemian collections from the National Gallery in Prague dating back to the 14th and 15th centuries. The design of the confocal setup allows its transport and the in-situ measurement of paintings in depositories or even in exhibition areas. The advantages of our confocal setup with movable collimating optics are also described.

  1. X-ray microscopes at BESSY II

    International Nuclear Information System (INIS)

    Guttmann, P.; Niemann, B.; Thieme, J.; Wiesemann, U.; Rudolph, D.; Schmahl, G.

    2000-01-01

    The undulator U41 at BESSY II will be used as source for X-ray microscopes. An overview of the X-ray microscopy area is presented. After finishing the construction phase a transmission X-ray microscope, a scanning transmission X-ray microscope and an X-ray test chamber will be available. The transmission X-ray microscope will allow investigations with high lateral resolution at moderate energy resolution while the scanning transmission X-ray microscope will allow high energy resolution at moderate lateral resolution of the same specimen

  2. Wolter x-ray microscope calibration

    International Nuclear Information System (INIS)

    Gerassimenko, M.

    1986-01-01

    A 22 x Wolter microscope was calibrated after several months of operation in the Lawrence Livermore National Laboratory (LLNL) Inertial Confinement Fusion program. Placing a point x-ray source at the microscope focus, I recorded the image plane spectrum, as well as the direct spectrum, and from the ratio of these two spectra derived an accurate estimate of the microscope solid angle in the 1-4 keV range. The solid angle was also calculated using the microscope geometry and composition. Comparison of this calculated value with the solid angle that was actually measured suggests contamination of the microscope surface

  3. Wolter x-ray microscope calibration

    International Nuclear Information System (INIS)

    Gerassimenko, M.

    1986-06-01

    A 22 x Wolter microscope was calibrated after several months of operation in the Lawrence Livermore National laboratory (LLNL) Inertial Confinement Fusion program. Placing a point x-ray source at the microscope focus, I recorded the image plane spectrum, as well as the direct spectrum, and from the ratio of these two spectra derived an accurate estimate of the microscope solid angle in the 1 to 4 keV range. The solid angle was also calculated using the microscope geometry and composition. Comparison of this calculated value with the solid angle that was actually measured suggests contamination of the microscope surface

  4. The illumination characteristics of operative microscopes.

    Science.gov (United States)

    Schutt, Christopher A; Redding, Brandon; Cao, Hui; Michaelides, Elias

    2015-01-01

    Modern operative microscopes use light sources which possess the power to severely damage underlying tissue. Currently, manufacturers provide a safety warning of this possibility. However, they are unable to suggest specific settings due to a stated "lack of scientific publications on this topic". We aim to radiometrically evaluate multiple otologic microscopes at variables which effect irradiance in order to determine reference emissions levels and provide guidelines for improved intraoperative safety. The optical radiance of four otologic microscopes was evaluated at variable field illumination sizes (spot size), intensity settings and working distances. The spectral emission of each microscope was separately measured. The energy absorbed in skin with representative properties was then calculated as a function of time for each microscope by accounting for the emission spectrum of the microscope and the absorption spectrum of skin. Microscopes showed a wide range of optical radiance based on model, spots size, intensity setting and working distances. Spectral emission of all four microscopes was centered in the visible spectrum with minimal ultraviolet or infrared contribution. A large amount of energy is absorbed by skin during usage of operative microscopes. The highest calculated absorption at 200 min of use was 736.26 J/cm(2). Operative microscopes have the ability to cause patient morbidity secondary to the energy they impart. In an effort to decrease potential injury we recommend that physicians be aware of their microscopes properties and how to control variables which effect irradiance of the skin. Published by Elsevier Inc.

  5. Robotic autopositioning of the operating microscope.

    Science.gov (United States)

    Oppenlander, Mark E; Chowdhry, Shakeel A; Merkl, Brandon; Hattendorf, Guido M; Nakaji, Peter; Spetzler, Robert F

    2014-06-01

    Use of the operating microscope has become pervasive since its introduction to the neurosurgical world. Neuronavigation fused with the operating microscope has allowed accurate correlation of the focal point of the microscope and its location on the downloaded imaging study. However, the robotic ability of the Pentero microscope has not been utilized to orient the angle of the microscope or to change its focal length to hone in on a predefined target. To report a novel technology that allows automatic positioning of the operating microscope onto a set target and utilization of a planned trajectory, either determined with the StealthStation S7 by using preoperative imaging or intraoperatively with the microscope. By utilizing the current motorized capabilities of the Zeiss OPMI Pentero microscope, a robotic autopositioning feature was developed in collaboration with Surgical Technologies, Medtronic, Inc. (StealthStation S7). The system is currently being tested at the Barrow Neurological Institute. Three options were developed for automatically positioning the microscope: AutoLock Current Point, Align Parallel to Plan, and Point to Plan Target. These options allow the microscope to pivot around the lesion, hover in a set plane parallel to the determined trajectory, or rotate and point to a set target point, respectively. Integration of automatic microscope positioning into the operative workflow has potential to increase operative efficacy and safety. This technology is best suited for precise trajectories and entry points into deep-seated lesions.

  6. Light Microsopy Module, International Space Station Premier Automated Microscope

    Science.gov (United States)

    Meyer, William V.; Sicker, Ronald J.; Chiaramonte, Francis P.; Brown, Daniel F.; O'Toole, Martin A.; Foster, William M.; Motil, Brian J.; Abbot-Hearn, Amber Ashley; Atherton, Arthur Johnson; Beltram, Alexander; hide

    2015-01-01

    The Light Microscopy Module (LMM) was launched to the International Space Station (ISS) in 2009 and began science operations in 2010. It continues to support Physical and Biological scientific research on ISS. During 2015, if all goes as planned, five experiments will be completed: [1] Advanced Colloids Experiments with a manual sample base -3 (ACE-M-3), [2] the Advanced Colloids Experiment with a Heated Base -1 (ACE-H-1), [3] (ACE-H-2), [4] the Advanced Plant Experiment -03 (APEX-03), and [5] the Microchannel Diffusion Experiment (MDE). Preliminary results, along with an overview of present and future LMM capabilities will be presented; this includes details on the planned data imaging processing and storage system, along with the confocal upgrade to the core microscope. [1] New York University: Paul Chaikin, Andrew Hollingsworth, and Stefano Sacanna, [2] University of Pennsylvania: Arjun Yodh and Matthew Gratale, [3] a consortium of universities from the State of Kentucky working through the Experimental Program to Stimulate Competitive Research (EPSCoR): Stuart Williams, Gerold Willing, Hemali Rathnayake, et al., [4] from the University of Florida and CASIS: Anna-Lisa Paul and Rob Ferl, and [5] from the Methodist Hospital Research Institute from CASIS: Alessandro Grattoni and Giancarlo Canavese.

  7. Microscopical advances in assisted reproduction.

    Science.gov (United States)

    Baccetti, B

    2004-01-01

    In a series of papers carried out by this laboratory it was demonstrated that the quality of sterile males sperm, assessed submicroscopically and mathematically, is closely correlated with the success of the various procedures of assisted reproduction. If we attempt to select hypothetically optimal spermatozoa destined to the ICSI by light inverted microscopy, a considerable amount of ultrastructural information is lost and our selection is merely based on the motility. In this study we apply polarization microscopy to the ICSI technique, introducing polarizing and analyzing lenses in an inverted microscope model, operating in a transparent container. The retardation of the birefringence in the various organelles is evaluated by compensators, and the images are transmitted to a video system, and stored in a computer. Spermatozoa are maintained alive and perfectly motile in this polarizing inverted microscope, and the character of the birefringence is the same as in fixed and sectioned biological material examined by polarization microscopy. The birefringence of the sperm structures allows a sperm analysis closer to TEM than to phase contrast light microscopy analysis.

  8. Mitochondrial behavior during oogenesis in zebrafish: a confocal microscopy analysis.

    Science.gov (United States)

    Zhang, Yong-Zhong; Ouyang, Ying-Chun; Hou, Yi; Schatten, Heide; Chen, Da-Yuan; Sun, Qing-Yuan

    2008-03-01

    The behavior of mitochondria during early oogenesis remains largely unknown in zebrafish. We used three mitochondrial probes (Mito Tracker Red CMXRos, Mito Tracker Green FM, and JC-1) to stain early zebrafish oocyte mitochondria, and confocal microscopy to analyze mitochondrial aggregation and distribution. By using fluorescence recovery after photobleaching (FRAP), we traced mitochondrial movement. The microtubule assembly inhibitor nocodazole and microfilament inhibitor cytochalasin B (CB) were used to analyze the role of microtubules and microfilaments on mitochondrial movement. By using the dual emission probe, JC-1, and oxidative phosphorylation uncoupler, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), we determined the distribution of active and inactive (low-active) mitochondria. Green/red fluorescence ratios of different sublocations in different oocyte groups stained by JC-1 were detected in merged (green and red) images. Our results showed that mitochondria exhibited a unique distribution pattern in early zebrafish oocytes. They tended to aggregate into large clusters in early stage I oocytes, but in a threadlike state in latter stage I oocytes. We detected a lower density mitochondrial area and a higher density mitochondrial area on opposite sides of the germinal vesicle. The green/red fluorescence ratios in different sublocations in normal oocytes were about 1:1. This implies that active mitochondria were distributed in all sublocations. FCCP treatment caused significant increases in the ratios. CB and nocodazole treatment caused an increase of the ratios in clusters and mitochondrial cloud, but not in dispersed areas. Mitochondria in different sublocations underwent fast dynamic movement. Inhibition or disruption of microtubules or microfilaments resulted in even faster mitochondrial free movement.

  9. Detection of desmoplastic melanoma with dermoscopy and reflectance confocal microscopy.

    Science.gov (United States)

    Maher, N G; Solinas, A; Scolyer, R A; Puig, S; Pellacani, G; Guitera, P

    2017-12-01

    Desmoplastic melanoma (DM) is frequently misdiagnosed clinically and often associated with melanoma in situ (MIS). To improve the detection of DM using dermoscopy and reflectance confocal microscopy (RCM). A descriptive analysis of DM dermoscopy features and a case-control study within a melanoma population for RCM feature evaluation was performed blindly, using data obtained between 2005 and 2015. After retrospectively identifying all DM cases with RCM data over the study period (n = 16), a control group of non-DM melanoma patients with RCM data, in a ratio of at least 3 : 1, was selected. The control group was matched by age and primary tumour site location, divided into non-DM invasive melanomas (n = 27) and MIS (n = 27). Invasive melanomas were selected according to the melanoma subtypes associated with the DM cases. The main outcomes were the frequency of melanoma-specific features on dermoscopy for DM; and the odds ratios of RCM features to distinguish DM from MIS and/or other invasive melanomas; or MIS from the combined invasive melanoma group. At least one of the 14 melanoma-specific features evaluated on dermoscopy was found in 100% of DMs (n = 15 DM with dermoscopy). Known RCM melanoma predictors were commonly found in the DMs, such as pagetoid cells (100%) and cell atypia (100%). The RCM feature of spindle cells in the superficial dermis was more common in DM compared with the entire melanoma control group (OR 3.82, 95% CI 1.01-14.90), and particularly compared to MIS (OR 5.48, 95% CI 1.11-32.36). Nucleated cells in the dermis and the RCM correlate of dermal inflammation were also significant RCM features favouring DM over MIS, as well as invasive melanoma over MIS. Dermoscopy and RCM may be useful tools for the identification of DM. Certain RCM features may help distinguish DM from MIS and other invasive melanomas. Larger studies are warranted. © 2017 European Academy of Dermatology and Venereology.

  10. 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.

  11. Reflectance confocal microscopy features of seborrheic dermatitis for plaque psoriasis differentiation.

    Science.gov (United States)

    Agozzino, Marina; Berardesca, Enzo; Donadio, Carlo; Franceschini, Chiara; de Felice, Catia Margarete; Cavallotti, Claudia; Sperduti, Isabella; Ardigò, Marco

    2014-01-01

    Plaque psoriasis (PP) and seborrheic dermatitis (SD) are chronic inflammatory skin diseases with similar clinical and pathological features. Differential diagnosis can be difficult, especially when particular skin areas of the face are involved. Reflectance confocal microscopy (RCM) has been demonstrated to be useful for 'real-time' diagnosis of skin inflammatory diseases. To define distinctive confocal criteria of SD and to evaluate the usefulness of this technique for noninvasive differential diagnosis with PP. A total of 40 patients affected by PP and 19 patients by SD involving the face were recruited and subjected to RCM evaluation. Univariate and adjusted odds ratios were calculated. Discriminant functions were used to plot ROC curves. The results disclosed specific patterns for SD and PP. The following distinctive confocal features for SD have been identified: spongiosis, dermal inflammation and horizontal orientation of dilated blood vessels. SD has a specific and easily recognizable confocal pattern supporting clinical differentiation with PP.

  12. Design of measurement system of 3D surface profile based on chromatic confocal technology

    Science.gov (United States)

    Wang, An-su; Xie, Bin; Liu, Zi-wei

    2018-01-01

    Chromatic confocal 3D profilometer has widely used in science investigation and industry fields recently for its high precision, great measuring range and numerical surface characteristic. It can provide exact and omnidirectional solution for manufacture and research by 3D non-contact surface analysis technique. The article analyzes the principle of surface measurement with chromatic confocal technology, and provides the designing indicators and requirements of the confocal system. As the key component, the dispersive objective used to achieve longitudinal focus vibration with wavelength was designed. The objective disperses the focus of wavelength between 400 700 nm to 15 mm longitudinal range. With selected spectrometer, the resolution of chromatic confocal 3D profilometer is no more than 5 μm, which can meet needs for the high precision non-contact surface profile measurement.

  13. 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

  14. Similar but Different: How Reflectance Confocal Microscopy May Help in the Diagnosis of Pink Lesions.

    Science.gov (United States)

    Ferrari, Federica; Bassoli, Sara; Pellacani, Giovanni; Argenziano, Giuseppe; Cesinaro, Anna Maria; Longo, Caterina

    2017-01-01

    Among skin neoplasms, solitary pink tumors represent challenging lesions in clinical practice since they can mimic melanocytic and nonmelanocytic lesions or even inflammatory ones. In this case series we described dermoscopic and confocal features of 2 couples of similar lesions in order to achieve the correct diagnosis and the best therapeutic approach. During clinical routine practice, 2 couples of clinically and dermoscopically similar lesions were examined by means of confocal microscopy. All lesions revealed no clear-cut diagnostic features on dermoscopy. However, confocal microscopy revealed tumor islands with palisading cells and a dark clefting at the periphery in basal cell carcinomas. In the other "false twin" lesions, atypical cells and elongated junctional nests were observed and the diagnosis of amelanotic melanomas was rendered. In the current case series, the combined use of dermoscopy and reflectance confocal microscopy was an optimal workup for difficult-to-diagnose lesions such as pink tumors. © 2017 S. Karger AG, Basel.

  15. Instant super-resolution imaging in live cells and embryos via analog image processing.

    Science.gov (United States)

    York, Andrew G; Chandris, Panagiotis; Nogare, Damian Dalle; Head, Jeffrey; Wawrzusin, Peter; Fischer, Robert S; Chitnis, Ajay; Shroff, Hari

    2013-11-01

    Existing super-resolution fluorescence microscopes compromise acquisition speed to provide subdiffractive sample information. We report an analog implementation of structured illumination microscopy that enables three-dimensional (3D) super-resolution imaging with a lateral resolution of 145 nm and an axial resolution of 350 nm at acquisition speeds up to 100 Hz. By using optical instead of digital image-processing operations, we removed the need to capture, store and combine multiple camera exposures, increasing data acquisition rates 10- to 100-fold over other super-resolution microscopes and acquiring and displaying super-resolution images in real time. Low excitation intensities allow imaging over hundreds of 2D sections, and combined physical and computational sectioning allow similar depth penetration to spinning-disk confocal microscopy. We demonstrate the capability of our system by imaging fine, rapidly moving structures including motor-driven organelles in human lung fibroblasts and the cytoskeleton of flowing blood cells within developing zebrafish embryos.

  16. Fluorescence live cell imaging.

    Science.gov (United States)

    Ettinger, Andreas; Wittmann, Torsten

    2014-01-01

    Fluorescence microscopy of live cells has become an integral part of modern cell biology. Fluorescent protein (FP) tags, live cell dyes, and other methods to fluorescently label proteins of interest provide a range of tools to investigate virtually any cellular process under the microscope. The two main experimental challenges in collecting meaningful live cell microscopy data are to minimize photodamage while retaining a useful signal-to-noise ratio and to provide a suitable environment for cells or tissues to replicate physiological cell dynamics. This chapter aims to give a general overview on microscope design choices critical for fluorescence live cell imaging that apply to most fluorescence microscopy modalities and on environmental control with a focus on mammalian tissue culture cells. In addition, we provide guidance on how to design and evaluate FP constructs by spinning disk confocal microscopy. © 2014 Elsevier Inc. All rights reserved.

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

    OpenAIRE

    NEAUPORT, Jérôme; Cormont, P; Legros, P; Ambard, C; Destribats, J

    2009-01-01

    International audience; 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...

  18. Multisource, Phase-controlled Radiofrequency for Treatment of Skin Laxity: Correlation Between Clinical and In-vivo Confocal Microscopy Results and Real-Time Thermal Changes.

    Science.gov (United States)

    Royo de la Torre, Josefina; Moreno-Moraga, Javier; Muñoz, Estefania; Cornejo Navarro, Paloma

    2011-01-01

    The objective of this study was to analyze the correlation between degrees of clinical improvement and microscopic changes detected using confocal microscopy at the temperature gradients reached in patients treated for skin laxity with a phase-controlled, multisource radiofrequency system. Patients with skin laxity in the abdominal area were treated in six sessions with radiofrequency (the first 4 sessions were held at 2-week intervals and the 2 remaining sessions at 3-week intervals). Patients attended monitoring at 6, 9, and 12 months. 33 patients (all women). The authors recorded the following: variations in weight, measurements of the contour of the treated area and control area, evaluation of clinical improvement by the clinician and by the patient, images taken using an infrared camera, temperature (before, immediately after, and 20 minutes after the procedure), and confocal microscopy images (before treatment and at 6, 9, and 12 months). The degree of clinical improvement was contrasted by two external observers (clinicians). The procedure was performed using a new phase-controlled, multipolar radiofrequency system. The results reveal a greater degree of clinical improvement in patients with surface temperature increases greater than 11.5ºC at the end of the procedure and remaining greater than 4.5ºC 20 minutes later. These changes induced by radiofrequency were contrasted with the structural improvements observed at the dermal-epidermal junction using confocal microscopy. Changes are more intense and are statistically correlated with patients who show a greater degree of improvement and have higher temperature gradients at the end of the procedure and 20 minutes later. Monitoring and the use of parameters to evaluate end-point values in skin quality treatment by multisource, phased-controlled radiofrequency can help optimize aesthetic outcome.

  19. Acute lung allograft rejection: diagnostic role of probe-based confocal laser endomicroscopy of the respiratory tract.

    Science.gov (United States)

    Yserbyt, Jonas; Dooms, Christophe; Decramer, Marc; Verleden, Geert M

    2014-05-01

    Acute cellular rejection (AR) after lung transplantation may result in significant morbidity and mortality both on the short and long term. Transbronchial biopsy through flexible bronchoscopy is highly sensitive for the diagnosis of AR, but reproducibility of histopathologic interpretation is less convincing. Probe-based confocal laser endomicroscopy (pCLE), a novel imaging tool in the field of respiratory medicine, enables real-time imaging of the pulmonary acini. We performed 105 bronchoscopies in lung transplant recipients, combining both transbronchial biopsies and pCLE. We conducted an observational survey for pCLE findings in AR. Calculations for cellularity showed a median cell count (ACA) of 50 (IQR 18 to 120) cells per microscopic field for AR and 10 (IQR 0 to 15) cells per microscopic field for matched controls (p = 0.0004). Cellular autofluorescence in the AR group was 1,163 (± 157) units and 489 (± 101) units for the matched controls (p = 0.0009). Autofluorescent cells were present in 73% (± 10) of the recorded frames in the AR group and in only 42% (± 9) of the recorded frames in the control group (p = 0.03). Contingency analysis for the presence/absence of ACA in the AR group versus the control group showed a sensitivity of 0.93 and a specificity of 0.46 (relative risk = 6.5 [95% CI 0.94 to 44.8], p = 0.01). The consecutive application of 3 pCLE criteria resulted in a sensitivity of 0.93 and a specificity of 0.83 for detection of AR. Our observational survey suggests the existence of specific pCLE characteristics in patients with AR. Further efforts are necessary to validate these findings prospectively. Copyright © 2014 International Society for Heart and Lung Transplantation. Published by Elsevier Inc. All rights reserved.

  20. Spinning disk test study on erase band and write width for shingled magnetic recording

    Science.gov (United States)

    Chandrasekaran, S.; Supnithi, P.; Warisarn, C.; Bai, D.

    2014-05-01

    Shingled magnetic recording (SMR) has been predicted as one of the novel technologies to extend the areal density beyond 1 Tb/in2. It was widely thought by many researchers that very wide writers could be used for SMR, as it involves corner writing and thus writability would no longer be an issue in SMR. However, in our experimental study, we reveal that there exists an optimal write width for a given writer design due to the erase band limitations. Also we propose the optimal write width for a given shingle track pitch condition.

  1. Acute shear stress direction dictates adherent cell remodeling and verifies shear profile of spinning disk assays

    International Nuclear Information System (INIS)

    Fuhrmann, Alexander; Engler, Adam J

    2015-01-01

    Several methods have been developed to quantify population level changes in cell attachment strength given its large heterogeneity. One such method is the rotating disk chamber or ‘spinning disk’ in which a range of shear forces are applied to attached cells to quantify detachment force, i.e. attachment strength, which can be heterogeneous within cell populations. However, computing the exact force vectors that act upon cells is complicated by complex flow fields and variable cell morphologies. Recent observations suggest that cells may remodel their morphology and align during acute shear exposure, but contrary to intuition, shear is not orthogonal to the radial direction. Here we theoretically derive the magnitude and direction of applied shear and demonstrate that cells, under certain physiological conditions, align in this direction within minutes. Shear force magnitude is also experimentally verified which validates that for spread cells shear forces and not torque or drag dominate in this assay, and demonstrates that the applied force per cell area is largely independent of initial morphology. These findings suggest that direct quantified comparison of the effects of shear on a wide array of cell types and conditions can be made with confidence using this assay without the need for computational or numerical modeling. (paper)

  2. Exophiala phaeomuriformis Fungal Keratitis: Case Report and In Vivo Confocal Microscopy Findings.

    Science.gov (United States)

    Aggarwal, Shruti; Yamaguchi, Takefumi; Dana, Reza; Hamrah, Pedram

    2017-03-01

    Corneal infections, particularly fungal keratitis due to rare fungal species, pose a diagnostic and therapeutic challenge because of difficulty in identification and varying susceptibility profiles. In this study, we report the first case of fungal keratitis because of Exophiala phaeomuriformis. We report the clinical findings and microbial identification techniques of a case of fungal keratitis due to E. phaeomuriformis. An 84-year-old woman presented with redness, pain, and itching in the left eye for 2 weeks. Slit-lamp biomicroscopy revealed one broken suture from previous penetrating keratoplasty (PKP), black infiltrates at the 4-o'clock position, without an overlying epithelial defect and hypopyon. Microbial identification was based cultures on Sabouraud dextrose agar and DNA sequencing and correlations to laser in vivo confocal microscopy (IVCM; Heidelberg Retinal Tomograph 3/Rostock Cornea Module, Heidelberg Engineering) and multiphoton microscopy (Ultima Microscope; Prairie Technologies) images. Slit-lamp biomicroscopy revealed one broken suture from previous PKP, black infiltrates at the 4-o'clock position, without an overlying epithelial defect and hypopyon. Based on a clinical suspicion of fungal keratitis, antifungals and fortified antibiotics were started. However, the patient did not respond to therapy and required urgent PKP. After surgery, the patient was maintained on topical and systemic voriconazole and also topical 2% cyclosporine for 5 months because of possibility of scleral involvement noticed during surgery. At the end of the treatment period, her vision improved from hand motion to 20/40, with no recurrence observed in a follow-up period of 1 year. Results of diagnostic tests were supported by fungal elements in stroma on IVCM. Culture from the infiltrate grew black yeast. DNA sequencing led to the diagnosis of E. phaeomuriformis keratitis. Antifungal susceptibility testing revealed sensitivity to voriconazole. This is, to our knowledge

  3. Focal Molography: Coherent Microscopic Detection of Biomolecular Interaction

    Directory of Open Access Journals (Sweden)

    Christof Fattinger

    2014-08-01

    . We discuss the implementation of a readout system for molograms on a thin-film optical waveguide by adapting a confocal laser-scanning microscope to a bifocal laser-scanning microscope.

  4. Handheld optical coherence tomography-reflectance confocal microscopy probe for detection of basal cell carcinoma and delineation of margins

    Science.gov (United States)

    Iftimia, Nicusor; Yélamos, Oriol; Chen, Chih-Shan J.; Maguluri, Gopi; Cordova, Miguel A.; Sahu, Aditi; Park, Jesung; Fox, William; Alessi-Fox, Christi; Rajadhyaksha, Milind

    2017-07-01

    We present a hand-held implementation and preliminary evaluation of a combined optical coherence tomography (OCT) and reflectance confocal microscopy (RCM) probe for detecting and delineating the margins of basal cell carcinomas (BCCs) in human skin in vivo. A standard OCT approach (spectrometer-based) with a central wavelength of 1310 nm and 0.11 numerical aperture (NA) was combined with a standard RCM approach (830-nm wavelength and 0.9 NA) into a common path hand-held probe. Cross-sectional OCT images and enface RCM images are simultaneously displayed, allowing for three-dimensional microscopic assessment of tumor morphology in real time. Depending on the subtype and depth of the BCC tumor and surrounding skin conditions, OCT and RCM imaging are able to complement each other, the strengths of each helping overcome the limitations of the other. Four representative cases are summarized, out of the 15 investigated in a preliminary pilot study, demonstrating how OCT and RCM imaging may be synergistically combined to more accurately detect BCCs and more completely delineate margins. Our preliminary results highlight the potential benefits of combining the two technologies within a single probe to potentially guide diagnosis as well as treatment of BCCs.

  5. Detecting protein complexes in living cells from laser scanning confocal image sequences by the cross correlation raster image spectroscopy method.

    Science.gov (United States)

    Digman, Michelle A; Wiseman, Paul W; Horwitz, Alan R; Gratton, Enrico

    2009-01-01

    We describe a general method for detecting molecular complexes based on the analysis of single molecule fluorescence fluctuations from laser scanning confocal images. The method detects and quantifies complexes of two different fluorescent proteins noninvasively in living cells. Because in a raster scanned image successive pixels are measured at different times, the spatial correlation of the image contains information about dynamic processes occurring over a large time range, from the microseconds to seconds. The correlation of intensity fluctuations measured simultaneously in two channels detects protein complexes that carry two molecules of different colors. This information is obtained from the entire image. A map of the spatial distribution of protein complexes in the cell and their diffusion and/or binding properties can be constructed. Using this cross correlation raster image spectroscopy method, specific locations in the cell can be visualized where dynamics of binding and unbinding of fluorescent protein complexes occur. This fluctuation imaging method can be applied to commercial laser scanning microscopes thereby making it accessible to a large community of scientists.

  6. In Vivo Confocal Fluorescence Imaging of the Intratumor Distribution of the Photosensitizer Mono-l-Aspartylchlorin-e6

    Directory of Open Access Journals (Sweden)

    Soumya Mitra

    2008-05-01

    Full Text Available We present an in vivo fluorescence microscopic evaluation of intratumor distribution of the photosensitizer mono-l-aspartylchlorin-e6 (NPe6 in an intradermal mouse EMT6 tumor model. Although the identification of favorable photophysical and pharmacological properties has led to the development of new photosensitizers in photodynamic therapy, their intratumor distribution kinetics have remained relatively understudied. In this study, we used confocal fluorescence microscopy to follow the transport of NPe6 in vivo after systemic administration through the tail vein. Labeling of vasculature using fluorophore-conjugated anti-CD31 antibodies allows visualization of the uptake of NPe6 in tumor and normal vessels and its partitioning kinetics into the adjacent parenchyma for 3 hours after injection. During the initial 60 minutes after injection, the drug is predominantly confined to the vasculature. Subsequently, it significantly redistributes throughout the extravascular regions with no discernable difference in its extravasation rate between tumor and normal tissues. Further, we investigate the sensitizer’s altered intratumor distribution in response to photodynamic therapy irradiation and observe that treatment-induced changes in vessel permeability caused enhanced accumulation of NPe6 in the extravascular space. Our findings are of immediate clinical relevance and demonstrate the importance of an in vivo imaging approach to examine the dynamic process of intratumor drug distribution.

  7. Design of a transmission electron positron microscope

    International Nuclear Information System (INIS)

    Doyama, Masao; Inoue, M.; Kogure, Y.; Hayashi, Y.; Yoshii, T.; Kurihara, T.; Tsuno, K.

    2003-01-01

    This paper reports the plans and design of positron-electron microscopes being built at KEK (High Energy Accelerator Research Organization), Tsukuba, Japan. A used electron microscope is altered. The kinetic energies of positrons produced by accelerators or by nuclear decays are not a unique value but show a spread over in a wide range. Positron beam is guided to a transmission electron microscope (JEM100SX). Positrons are moderated by a tungsten foil, are accelerated and are focused on a nickel sheet. The monochromatic focused beam is injected into an electron microscope. The focusing and aberration of positrons are the same as electrons in a magnetic system which are used in commercial electron microscopes. Imaging plates are used to record positron images for the transmission electron microscope. (author)

  8. Testing microscopes between market and scientific strategies

    OpenAIRE

    Ratcliff, Marc

    2007-01-01

    This paper claims that the testing of microscopes during the eighteenth century reveals specific types of interaction between makers and users and links between scientific and economic interests. Basic procedures for the comparison and test of microscopes existed already in the Enlightenment although many historians thought that these were invented during the nineteenth century. The paper discusses three kinds of tests, advertising, the admission of a microscope in the laboratory, and finally...

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

    Science.gov (United States)

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

    2015-01-01

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

  10. Calibrated atomic force microscope measurements of vickers hardness indentations and tip production and characterisation for scanning tunelling microscope

    DEFF Research Database (Denmark)

    Jensen, Carsten P.

    Calibrated atomic force microscope measurements of vickers hardness indentations and tip production and characterisation for scanning tunelling microscope......Calibrated atomic force microscope measurements of vickers hardness indentations and tip production and characterisation for scanning tunelling microscope...

  11. Microscopic description of magnetized plasma: quasiparticle concept

    International Nuclear Information System (INIS)

    Sosenko, P.P.; Decyk, V.K.

    1993-01-01

    A quasiparticle concept is developed systematically, from first principles, within the context of microscopic description of magnetized plasma. It is argued that the zeroth velocity-gyroangle harmonic of the microscopic particle distribution function under the gyrokinetic change of variables can be taken as a microscopic quasi-particle density in a reduced phase space. The nature of quasiparticles is discussed and equations of their motion are derived within both exact and reduced microscopic descriptions. The reduced one employs explicitly the separation of interesting time scales. (orig.)

  12. Foldscope: origami-based paper microscope.

    Directory of Open Access Journals (Sweden)

    James S Cybulski

    Full Text Available Here we describe an ultra-low-cost origami-based approach for large-scale manufacturing of microscopes, specifically demonstrating brightfield, darkfield, and fluorescence microscopes. Merging principles of optical design with origami enables high-volume fabrication of microscopes from 2D media. Flexure mechanisms created via folding enable a flat compact design. Structural loops in folded paper provide kinematic constraints as a means for passive self-alignment. This light, rugged instrument can survive harsh field conditions while providing a diversity of imaging capabilities, thus serving wide-ranging applications for cost-effective, portable microscopes in science and education.

  13. A microscope for Fermi gases

    International Nuclear Information System (INIS)

    Omran, Ahmed

    2016-01-01

    This thesis reports on a novel quantum gas microscope to investigate many-body systems of fermionic atoms in optical lattices. Single-site resolved imaging of ultracold lattice gases has enabled powerful studies of bosonic quantum many-body systems. The extension of this capability to Fermi gases offers new prospects to studying complex phenomena of strongly correlated systems, for which numerical simulations are often out of reach. Using standard techniques of laser cooling, optical trapping, and evaporative cooling, ultracold Fermi gases of 6 Li are prepared and loaded into a large-scale 2D optical lattice of flexible geometry. The atomic distribution is frozen using a second, short-scaled lattice, where we perform Raman sideband cooling to induce fluorescence on each atom while maintaining its position. Together with high-resolution imaging, the fluorescence signals allow for reconstructing the initial atom distribution with single-site sensitivity and high fidelity. Magnetically driven evaporative cooling in the plane allows for producing degenerate Fermi gases with almost unity filling in the initial lattice, allowing for the first microscopic studies of ultracold gases with clear signatures of Fermi statistics. By preparing an ensemble of spin-polarised Fermi gases, we detect a flattening of the density profile towards the centre of the cloud, which is a characteristic of a band-insulating state. In one set of experiments, we demonstrate that losses of atom pairs on a single lattice site due to light-assisted collisions are circumvented. The oversampling of the second lattice allows for deterministic separation of the atom pairs into different sites. Compressing a high-density sample in a trap before loading into the lattice leads to many double occupancies of atoms populating different bands, which we can image with no evidence for pairwise losses. We therefore gain direct access to the true number statistics on each lattice site. Using this feature, we can

  14. Scanning Microscopes Using X Rays and Microchannels

    Science.gov (United States)

    Wang, Yu

    2003-01-01

    Scanning microscopes that would be based on microchannel filters and advanced electronic image sensors and that utilize x-ray illumination have been proposed. Because the finest resolution attainable in a microscope is determined by the wavelength of the illumination, the xray illumination in the proposed microscopes would make it possible, in principle, to achieve resolutions of the order of nanometers about a thousand times as fine as the resolution of a visible-light microscope. Heretofore, it has been necessary to use scanning electron microscopes to obtain such fine resolution. In comparison with scanning electron microscopes, the proposed microscopes would likely be smaller, less massive, and less expensive. Moreover, unlike in scanning electron microscopes, it would not be necessary to place specimens under vacuum. The proposed microscopes are closely related to the ones described in several prior NASA Tech Briefs articles; namely, Miniature Microscope Without Lenses (NPO-20218), NASA Tech Briefs, Vol. 22, No. 8 (August 1998), page 43; and Reflective Variants of Miniature Microscope Without Lenses (NPO-20610), NASA Tech Briefs, Vol. 26, No. 9 (September 2002) page 6a. In all of these microscopes, the basic principle of design and operation is the same: The focusing optics of a conventional visible-light microscope are replaced by a combination of a microchannel filter and a charge-coupled-device (CCD) image detector. A microchannel plate containing parallel, microscopic-cross-section holes much longer than they are wide is placed between a specimen and an image sensor, which is typically the CCD. The microchannel plate must be made of a material that absorbs the illuminating radiation reflected or scattered from the specimen. The microchannels must be positioned and dimensioned so that each one is registered with a pixel on the image sensor. Because most of the radiation incident on the microchannel walls becomes absorbed, the radiation that reaches the

  15. Quantitative 3D elemental analysis inside plant roots by means of synchrotron confocal micro X-ray fluorescence

    Science.gov (United States)

    Terzano, R.; Vekemans, B.; Tomasi, N.; Spagnuolo, M.; Schoonjans, T.; Vincze, L.; Pinton, R.; Cesco, S.; Ruggiero, P.

    2009-04-01

    The knowledge of the distribution and concentration of elements within plants is a fundamental step to better understand how these plants uptake specific elements from the medium of growth and how they manage acquisition and compartmentalisation of nutrients as well as toxic metals. For some elements, either nutrients or toxicants, it can be of relevance to know their concentration level within microscopic volumes in plant organs, where they are stored or accumulated. Usually, this type of microscopic analysis requires complex cutting procedures and extensive sample manipulations. In this research, the technique of synchrotron micro X-ray fluorescence in the confocal mode was applied to image the distribution of elements in selected key-planes of tomato roots without the need of any sample preparation, except washing and freeze-drying. Using this method, a first polycapillary lens focussed the X-ray beam with an energy of 12.4 keV down to a 20 µm beam that is penetrating the sample, and a second polycapillary half-lens, that was positioned at the detection side at 90 degrees to the first polycapillary, could then restrict further the view on this irradiated volume to a defined microscopic volume (typically 20x20x20 µm3) from which the induced fluorescent radiation is finally collected by the energy dispersive detector. In this way, it was possible to investigate the concentration levels of some elements such as K, Ca, Mn, Fe, Cu and Zn within the roots of tomato plants. The quantification was performed by means of a dedicated XRF Fundamental Parameter (FP) method in order to calculate the concentrations of trace elements within the analysed plants. Utilizing fundamental atomic parameters, the applied FP method is taking into account the influence of sample self-absorption and especially the specific detection processes by the polycapillary lens. Quantification was assessed and validated by using different standards: NIST SRM 1573a (trace elements in tomato leaves

  16. A transmission positron microscope and a scanning positron microscope being built at KEK, Japan

    International Nuclear Information System (INIS)

    Doyama, M.; Inoue, M.; Kogure, Y.; Kurihara, T.; Yagishita, A.; Shidara, T.; Nakahara, K.; Hayashi, Y.; Yoshiie, T.

    2001-01-01

    This paper reports the plans of positron microscopes being built at KEK (High Energy Accelerator Research Organization), Tsukuba, Japan improving used electron microscopes. The kinetic energies of positron produced by accelerators or by nuclear decays have not a unique value but show a spread over in a wide range. Positron beam will be guided near electron microscopes, a transmission electron microscope (JEM100S) and a scanning electron microscope (JSM25S). Positrons are slowed down by a tungsten foil, accelerated and focused on a nickel sheet. The monochromatic focused beam will be injected into an electron microscope. The focusing of positrons and electrons is achieved by magnetic system of the electron microscopes. Imaging plates are used to record positron images for the transmission electron microscope. (orig.)

  17. Spectral Interferometry with Electron Microscopes.

    Science.gov (United States)

    Talebi, Nahid

    2016-09-21

    Interference patterns are not only a defining characteristic of waves, but also have several applications; characterization of coherent processes and holography. Spatial holography with electron waves, has paved the way towards space-resolved characterization of magnetic domains and electrostatic potentials with angstrom spatial resolution. Another impetus in electron microscopy has been introduced by ultrafast electron microscopy which uses pulses of sub-picosecond durations for probing a laser induced excitation of the sample. However, attosecond temporal resolution has not yet been reported, merely due to the statistical distribution of arrival times of electrons at the sample, with respect to the laser time reference. This is however, the very time resolution which will be needed for performing time-frequency analysis. These difficulties are addressed here by proposing a new methodology to improve the synchronization between electron and optical excitations through introducing an efficient electron-driven photon source. We use focused transition radiation of the electron as a pump for the sample. Due to the nature of transition radiation, the process is coherent. This technique allows us to perform spectral interferometry with electron microscopes, with applications in retrieving the phase of electron-induced polarizations and reconstructing dynamics of the induced vector potential.

  18. Hamiltonian mechanics limits microscopic engines

    Science.gov (United States)

    Anglin, James; Gilz, Lukas; Thesing, Eike

    2015-05-01

    We propose a definition of fully microscopic engines (micro-engines) in terms of pure mechanics, without reference to thermodynamics, equilibrium, or cycles imposed by external control, and without invoking ergodic theory. This definition is pragmatically based on the observation that what makes engines useful is energy transport across a large ratio of dynamical time scales. We then prove that classical and quantum mechanics set non-trivial limits-of different kinds-on how much of the energy that a micro-engine extracts from its fuel can be converted into work. Our results are not merely formal; they imply manageable design constraints on micro-engines. They also suggest the novel possibility that thermodynamics does not emerge from mechanics in macroscopic regimes, but rather represents the macroscopic limit of a generalized theory, valid on all scales, which governs the important phenomenon of energy transport across large time scale ratios. We propose experimental realizations of the dynamical mechanisms we identify, with trapped ions and in Bose-Einstein condensates (``motorized bright solitons'').

  19. Mice embryology: a microscopic overview.

    Science.gov (United States)

    Salvadori, Maria Letícia Baptista; Lessa, Thais Borges; Russo, Fabiele Baldino; Fernandes, Renata Avancini; Kfoury, José Roberto; Braga, Patricia Cristina Baleeiro Beltrão; Miglino, Maria Angélica

    2012-10-01

    In this work, we studied the embryology of mice of 12, 14, and 18 days of gestation by gross observation, light microscopy, and scanning electron microscopy. Grossly, the embryos of 12 days were observed in C-shaped region of the brain, eye pigmentation of the retina, first, second, and third pharyngeal arches gill pit nasal region on the fourth ventricle brain, cervical curvature, heart, liver, limb bud thoracic, spinal cord, tail, umbilical cord, and place of the mesonephric ridge. Microscopically, the liver, cardiovascular system and spinal cord were observed. In the embryo of 14 days, we observed structures that make up the liver and heart. At 18 days of gestation fetuses, it was noted the presence of eyes, mouth, and nose in the cephalic region, chest and pelvic region with the presence of well-developed limbs, umbilical cord, and placenta. Scanning electron microscopy in 18 days of gestation fetuses evidenced head, eyes closed eyelids, nose, vibrissae, forelimb, heart, lung, kidney, liver, small bowel, diaphragm, and part of the spine. The results obtained in this work describe the internal and external morphology of mice, provided by an integration of techniques and review of the morphological knowledge of the embryonic development of this species, as this animal is of great importance to scientific studies. Copyright © 2012 Wiley Periodicals, Inc.

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

    Science.gov (United States)

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

    2014-01-01

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

  1. Photon event distribution sampling: an image formation technique for scanning microscopes that permits tracking of sub-diffraction particles with high spatial and temporal resolutions.

    Science.gov (United States)

    Larkin, J D; Publicover, N G; Sutko, J L

    2011-01-01

    In photon event distribution sampling, an image formation technique for scanning microscopes, the maximum likelihood position of origin of each detected photon is acquired as a data set rather than binning photons in pixels. Subsequently, an intensity-related probability density function describing the uncertainty associated with the photon position measurement is applied to each position and individual photon intensity distributions are summed to form an image. Compared to pixel-based images, photon event distribution sampling images exhibit increased signal-to-noise and comparable spatial resolution. Photon event distribution sampling is superior to pixel-based image formation in recognizing the presence of structured (non-random) photon distributions at low photon counts and permits use of non-raster scanning patterns. A photon event distribution sampling based method for localizing single particles derived from a multi-variate normal distribution is more precise than statistical (Gaussian) fitting to pixel-based images. Using the multi-variate normal distribution method, non-raster scanning and a typical confocal microscope, localizations with 8 nm precision were achieved at 10 ms sampling rates with acquisition of ~200 photons per frame. Single nanometre precision was obtained with a greater number of photons per frame. In summary, photon event distribution sampling provides an efficient way to form images when low numbers of photons are involved and permits particle tracking with confocal point-scanning microscopes with nanometre precision deep within specimens. © 2010 The Authors Journal of Microscopy © 2010 The Royal Microscopical Society.

  2. Multibeam scanning electron microscope : Experimental results

    NARCIS (Netherlands)

    Mohammadi-Gheidari, A.; Hagen, C.W.; Kruit, P.

    2010-01-01

    The authors present the first results obtained with their multibeam scanning electron microscope. For the first time, they were able to image 196 (array of 14×14) focused beams of a multielectron beam source on a specimen using single beam scanning electron microscope (SEM) optics. The system

  3. Major Components of the Light Microscope

    OpenAIRE

    Centonze Frohlich, Victoria

    2008-01-01

    The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for different applications, and how it should be maintained as required to obtain maximum image-forming capacity and resolution. The components of the microscope are described in detail here.

  4. Major components of the light microscope.

    Science.gov (United States)

    Centonze Frohlich, Victoria

    2008-07-30

    The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for different applications, and how it should be maintained as required to obtain maximum image-forming capacity and resolution. The components of the microscope are described in detail here.

  5. Microscopic colitis : an unfamiliar but treatable disease

    NARCIS (Netherlands)

    van der Wouden, E. J.; Karrenbeld, A.; Kleibeuker, J. H.; Dijkstra, G.

    Chronic diarrhoea is a frequent complaint in clinical practice. Microscopic colitis is the cause of this symptom in 10% of these cases and the prevalence is rising. To exclude microscopic colitis a colonoscopy with multiple biopsies of different regions of the colon is mandatory. A sigmoidoscopy

  6. A microscopic model of triangular arbitrage

    Science.gov (United States)

    Aiba, Yukihiro; Hatano, Naomichi

    2006-11-01

    We introduce a microscopic model which describes the dynamics of each dealer in multiple foreign exchange markets, taking account of the triangular arbitrage transaction. The model reproduces the interaction among the markets well. We explore the relation between the parameters of the present microscopic model and the spring constant of a macroscopic model that we proposed previously.

  7. Quantum theory and microscopic mechanics. I

    International Nuclear Information System (INIS)

    Yussouff, M.

    1984-08-01

    The need for theoretical descriptions and experimental observations on 'small' individual systems is emphasized. It is shown that the mathematical basis for microscopic mechanics is very simple in one dimension. The square well problem is discussed to clarify general points about stationary states and the continuity of (p'/p) across potential boundaries in the applications of microscopic mechanics. (author)

  8. In situ protein expression in tumour spheres: development of an immunostaining protocol for confocal microscopy

    International Nuclear Information System (INIS)

    Weiswald, Louis-Bastien; Guinebretière, Jean-Marc; Richon, Sophie; Bellet, Dominique; Saubaméa, Bruno; Dangles-Marie, Virginie

    2010-01-01

    Multicellular tumour sphere models have been shown to closely mimic phenotype characteristics of in vivo solid tumours, or to allow in vitro propagation of cancer stem cells (CSCs). CSCs are usually characterized by the expression of specific membrane markers using flow cytometry (FC) after enzymatic dissociation. Consequently, the spatial location of positive cells within spheres is not documented. Confocal microscopy is the best technique for the imaging of thick biological specimens after multi-labelling but suffers from poor antibody penetration. Thus, we describe here a new protocol for in situ confocal imaging of protein expression in intact spheroids. Protein expression in whole spheroids (150 μm in diameter) from two human colon cancer cell lines, HT29 and CT320X6, has been investigated with confocal immunostaining, then compared with profiles obtained through paraffin immunohistochemistry (pIHC) and FC. Target antigens, relevant for colon cancer and with different expression patterns, have been studied. We first demonstrate that our procedure overcomes the well-known problem of antibody penetration in compact structures by performing immunostaining of EpCAM, a membrane protein expressed by all cells within our spheroids. EpCAM expression is detected in all cells, even the deepest ones. Likewise, antibody access is confirmed with CK20 and CD44 immunostaining. Confocal imaging shows that 100% of cells express β-catenin, mainly present in the plasma membrane with also cytoplasmic and nuclear staining, in agreement with FC and pIHC data. pIHC and confocal imaging show similar CA 19-9 cytoplasmic and membranar expression profile in a cell subpopulation. CA 19-9 + cell count confirms confocal imaging as a highly sensitive method (75%, 62% and 51%, for FC, confocal imaging and pIHC, respectively). Finally, confocal imaging reveals that the weak expression of CD133, a putative colon CSC marker, is restricted to the luminal cell surface of colorectal cancer acini

  9. Polymerized LB Films Imaged with a Combined Atomic Force Microscope-Fluorescence Microscope

    NARCIS (Netherlands)

    Putman, C.A.J.; Putman, Constant A.J.; Hansma, Helen G.; Gaub, Hermann E.; Hansma, Paul K.

    1992-01-01

    The first results obtained with a new stand-alone atomic force microscope (AFM) integrated with a standard Zeiss optical fluorescence microscope are presented. The optical microscope allows location and selection of objects to be imaged with the high-resolution AFM. Furthermore, the combined

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

    Science.gov (United States)

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

    2008-02-01

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

  11. Depth elemental imaging of forensic samples by confocal micro-XRF method.

    Science.gov (United States)

    Nakano, Kazuhiko; Nishi, Chihiro; Otsuki, Kazunori; Nishiwaki, Yoshinori; Tsuji, Kouichi

    2011-05-01

    Micro-XRF is a significant tool for the analysis of small regions. A micro-X-ray beam can be created in the laboratory by various focusing X-ray optics. Previously, nondestructive 3D-XRF analysis had not been easy because of the high penetration of fluorescent X-rays emitted into the sample. A recently developed confocal micro-XRF technique combined with polycapillary X-ray lenses enables depth-selective analysis. In this paper, we applied a new tabletop confocal micro-XRF system to analyze several forensic samples, that is, multilayered automotive paint fragments and leather samples, for use in the criminaliztics. Elemental depth profiles and mapping images of forensic samples were successfully obtained by the confocal micro-XRF technique. Multilayered structures can be distinguished in forensic samples by their elemental depth profiles. However, it was found that some leather sheets exhibited heterogeneous distribution. To confirm the validity, the result of a conventional micro-XRF of the cross section was compared with that of the confocal micro-XRF. The results obtained by the confocal micro-XRF system were in approximate agreement with those obtained by the conventional micro-XRF. Elemental depth imaging was performed on the paint fragments and leather sheets to confirm the homogeneity of the respective layers of the sample. The depth images of the paint fragment showed homogeneous distribution in each layer expect for Fe and Zn. In contrast, several components in the leather sheets were predominantly localized.

  12. An Interactive Visualization Tool for Multi-channel Confocal Microscopy Data in Neurobiology Research

    Science.gov (United States)

    Wan, Yong; Otsuna, Hideo; Chien, Chi-Bin; Hansen, Charles

    2010-01-01

    Confocal microscopy is widely used in neurobiology for studying the three-dimensional structure of the nervous system. Confocal image data are often multi-channel, with each channel resulting from a different fluorescent dye or fluorescent protein; one channel may have dense data, while another has sparse; and there are often structures at several spatial scales: subneuronal domains, neurons, and large groups of neurons (brain regions). Even qualitative analysis can therefore require visualization using techniques and parameters fine-tuned to a particular dataset. Despite the plethora of volume rendering techniques that have been available for many years, the techniques standardly used in neurobiological research are somewhat rudimentary, such as looking at image slices or maximal intensity projections. Thus there is a real demand from neurobiologists, and biologists in general, for a flexible visualization tool that allows interactive visualization of multi-channel confocal data, with rapid fine-tuning of parameters to reveal the three-dimensional relationships of structures of interest. Together with neurobiologists, we have designed such a tool, choosing visualization methods to suit the characteristics of confocal data and a typical biologist’s workflow. We use interactive volume rendering with intuitive settings for multidimensional transfer functions, multiple render modes and multi-views for multi-channel volume data, and embedding of polygon data into volume data for rendering and editing. As an example, we apply this tool to visualize confocal microscopy datasets of the developing zebrafish visual system. PMID:19834225

  13. Association between dermoscopic and reflectance confocal microscopy features of cutaneous melanoma with BRAF mutational status.

    Science.gov (United States)

    Bombonato, C; Ribero, S; Pozzobon, F C; Puig-Butille, J A; Badenas, C; Carrera, C; Malvehy, J; Moscarella, E; Lallas, A; Piana, S; Puig, S; Argenziano, G; Longo, C

    2017-04-01

    Melanomas harbouring common genetic mutations might share certain morphological features detectable with dermoscopy and reflectance confocal microscopy. BRAF mutational status is crucial for the management of metastatic melanoma. To correlate the dermoscopic characteristics of primary cutaneous melanomas with BRAF mutational status. Furthermore, a subset of tumours has also been analysed for the presence of possible confocal features that might be linked with BRAF status. Retrospectively acquired dermoscopic and confocal images of patients with melanoma in tertiary referral academic centres: Skin Cancer Unit in Reggio Emilia and at the Melanoma Unit in Barcelona. Kruskal-Wallis test, logistic regressions, univariate and multivariate analyses have been performed to find dermoscopic and confocal features significantly correlated with BRAF mutational status. Dermoscopically, the presence of irregular peripheral streaks and ulceration were positive predictors of BRAF-mutated melanomas with a statistically significance value, while dotted vessels were more represented in wild-type melanomas. None of the evaluated reflectance confocal microscopy features were correlated with genetic profiling. Ulceration and irregular peripheral streaks represent dermoscopic feature indicative for BRAF-mutated melanoma, while dotted vessels are suggestive for wild-type melanoma. © 2016 European Academy of Dermatology and Venereology.

  14. 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.

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

    International Nuclear Information System (INIS)

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

    2008-01-01

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

  16. An interactive visualization tool for multi-channel confocal microscopy data in neurobiology research

    KAUST Repository

    Yong Wan,

    2009-11-01

    Confocal microscopy is widely used in neurobiology for studying the three-dimensional structure of the nervous system. Confocal image data are often multi-channel, with each channel resulting from a different fluorescent dye or fluorescent protein; one channel may have dense data, while another has sparse; and there are often structures at several spatial scales: subneuronal domains, neurons, and large groups of neurons (brain regions). Even qualitative analysis can therefore require visualization using techniques and parameters fine-tuned to a particular dataset. Despite the plethora of volume rendering techniques that have been available for many years, the techniques standardly used in neurobiological research are somewhat rudimentary, such as looking at image slices or maximal intensity projections. Thus there is a real demand from neurobiologists, and biologists in general, for a flexible visualization tool that allows interactive visualization of multi-channel confocal data, with rapid fine-tuning of parameters to reveal the three-dimensional relationships of structures of interest. Together with neurobiologists, we have designed such a tool, choosing visualization methods to suit the characteristics of confocal data and a typical biologist\\'s workflow. We use interactive volume rendering with intuitive settings for multidimensional transfer functions, multiple render modes and multi-views for multi-channel volume data, and embedding of polygon data into volume data for rendering and editing. As an example, we apply this tool to visualize confocal microscopy datasets of the developing zebrafish visual system.

  17. Continuous-Wave Stimulated Emission Depletion Microscope for Imaging Actin Cytoskeleton in Fixed and Live Cells

    Directory of Open Access Journals (Sweden)

    Bhanu Neupane

    2015-09-01

    Full Text Available Stimulated emission depletion (STED microscopy provides a new opportunity to study fine sub-cellular structures and highly dynamic cellular processes, which are challenging to observe using conventional optical microscopy. Using actin as an example, we explored the feasibility of using a continuous wave (CW-STED microscope to study the fine structure and dynamics in fixed and live cells. Actin plays an important role in cellular processes, whose functioning involves dynamic formation and reorganization of fine structures of actin filaments. Frequently used confocal fluorescence and STED microscopy dyes were employed to image fixed PC-12 cells (dyed with phalloidin- fluorescein isothiocyante and live rat chondrosarcoma cells (RCS transfected with actin-green fluorescent protein (GFP. Compared to conventional confocal fluorescence microscopy, CW-STED microscopy shows improved spatial resolution in both fixed and live cells. We were able to monitor cell morphology changes continuously; however, the number of repetitive analyses were limited primarily by the dyes used in these experiments and could be improved with the use of dyes less susceptible to photobleaching. In conclusion, CW-STED may disclose new information for biological systems with a proper characteristic length scale. The challenges of using CW-STED microscopy to study cell structures are discussed.

  18. Comparative study of image contrast in scanning electron microscope and helium ion microscope.

    Science.gov (United States)

    O'Connell, R; Chen, Y; Zhang, H; Zhou, Y; Fox, D; Maguire, P; Wang, J J; Rodenburg, C

    2017-12-01

    Images of Ga + -implanted amorphous silicon layers in a 110 n-type silicon substrate have been collected by a range of detectors in a scanning electron microscope and a helium ion microscope. The effects of the implantation dose and imaging parameters (beam energy, dwell time, etc.) on the image contrast were investigated. We demonstrate a similar relationship for both the helium ion microscope Everhart-Thornley and scanning electron microscope Inlens detectors between the contrast of the images and the Ga + density and imaging parameters. These results also show that dynamic charging effects have a significant impact on the quantification of the helium ion microscope and scanning electron microscope contrast. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  19. Microscopic Study of Surface Microtopographic Characteristics of Dental Implants

    Science.gov (United States)

    Sezin, M.; Croharé, L.; Ibañez, J.C.

    2016-01-01

    Objective: To determine and compare the micro topographic characteristics of dental implants submitted to different surface treatments, using scanning electron microscopy (SEM). Materials and Methods: Implants were divided into 7 groups of 3 specimens each, according to the surface treatment used: group 1: Osseotite, BIOMET 3i; group 2: SLA surface, Institut Straumann AG; group 3: Oxalife surface, Tree-Oss implant; group 4: B&W implant surface; group 5: Q-implant surface; group 6: ML implant surface; group 7: RBM surface, Rosterdent implant. The surfaces were examined under SEM (Carl Zeiss FE-SEM-SIGMA). Image Proplus software was used to determine the number and mean diameter of pores per area unit (mm). The data obtained were analyzed with the Mann-Whitney test. A confocal laser microscope (LEXT-OLS4100 Olympus) was used to conduct the comparative study of surface roughness (Ra). Data were analyzed using Tukey's HSD test. Results: The largest average pore diameter calculated in microns was found in group 5 (3.45 µm+/-1.91) while the smallest in group 7 (1.47µm+/-1.29). Significant differences were observed among each one of the groups studied (p<0.05). The largest number of pores/mm2 was found in group 2 (229343) and the smallest number in group 4 (10937). Group 2 showed significant differences regarding the other groups (p<0.05). The greatest roughness (Ra) was observed in group 2 (0.975µm+/-0.115) and the smallest in group 4 (0.304µm+/-0.063). Group 2 was significantly different from the other groups (p<0.05). Conclusion: The micro topography observed in the different groups presented dissimilar and specific features, depending on the chemical treatment used for the surfaces.. PMID:27335615

  20. In-vivo confocal microscopy based on the Texas Instruments digital micromirror device

    Science.gov (United States)

    Botvinick, Elliott L.; Li, Florence; Cha, Sungdo; Gough, David A.; Fainman, Yeshaiahu; Price, Jeffrey H.

    2000-04-01

    Confocal microscopy is used widely for 3D biological imaging, but can be too slow for many applications. The limitations arise from scanning a single spot across the specimen at high speeds. Singe-spot confocal imaging usually works at a 1-2 Hz frame rate and faster systems tend to be signal/noise limited. Many live cellular events require both high speed and high SNR. Some parallel confocal systems have been developed to collect light from many points simultaneously to obtain high SNR and/or speed. Nipkow disks are compared of many pinholes, but have a fixed pattern and low light efficiency. Slit scanning systems collect an entire video line at a time, but compromise resolution. The TI Digital Micromirror Device (offers an alternative via large arrays of rapidly re-configurable micromirrors that can form arrays of reflection 'pinholes'. The prototype presented here exhibited 0.4 X 0.4 X 0.8 micrometers 3 resolution with a 100X 0.90 NA objective. An array of 10,000 or more neighborhoods, each compromising a single ON mirror in a group of OFF mirrors, creates the confocal parallelism. Alternating the ON mirror in each neighborhood until the image is completely formed on the CCD sensor enables transverse scanning. With 10,000-fold parallelism, for example, light collection efficiency and frame rate can both be 100X higher than in typical spot scanning. The high sensitivity allows high-speed confocal imaging at intensities below the cellular fluorotoxicity threshold. This was demonstrated in a hamster window preparation scanned daily in one-week longitudinal studies. Vessel geometry and localized blood flow were reconstructed to measure perfusion. High frame rate and sensitivity allowed real-time visualization of DiI stained intravascular red blood cells with no apparent tissue damage, supporting the tremendous potential advantages over current confocal technologies.

  1. Development and applications of the positron microscope

    International Nuclear Information System (INIS)

    1991-01-01

    Progress on the positron microscope during the past year has been steady, and we currently project that initial microscope images can be collected during mid to late summer of 1992. Work during the year has mainly been divided among four areas of effort: hardware construction; power supply and control system development; radioactive source fabrication; and planning of initial experimental projects. Details of progress in these areas will be given below. An initial optical design of the microscope was completed during 1990, but during the past year, significant improvements have been made to this design, and several limiting cases of microscope performance have been evaluated. The results of these evaluations have been extremely encouraging, giving us strong indications that the optical performance of the microscope will be better than originally anticipated. In particular, we should be able to explore ultimate performance capabilities of positron microscopy using our currently planned optical system, with improvements only in the image detector system, and the positron-source/moderator configuration. We should be able to study imaging reemission microscopy with resolutions approaching 10 Angstrom and be able to produce beam spots for rastered microscope work with diameters below the 1000 Angstrom diffusion limit. Because of these exciting new possibilities, we have decided to upgrade several microscope subsystems to levels consistent with ultimate performance earlier in our construction schedule than we had previously intended. In particular, alignment facilities in the optical system, vibration isolation, and power supply and control system flexibility have all been upgraded in their design over the past year

  2. Atomic Force Microscope Mediated Chromatography

    Science.gov (United States)

    Anderson, Mark S.

    2013-01-01

    The atomic force microscope (AFM) is used to inject a sample, provide shear-driven liquid flow over a functionalized substrate, and detect separated components. This is demonstrated using lipophilic dyes and normal phase chromatography. A significant reduction in both size and separation time scales is achieved with a 25-micron-length column scale, and one-second separation times. The approach has general applications to trace chemical and microfluidic analysis. The AFM is now a common tool for ultra-microscopy and nanotechnology. It has also been demonstrated to provide a number of microfluidic functions necessary for miniaturized chromatography. These include injection of sub-femtoliter samples, fluidic switching, and sheardriven pumping. The AFM probe tip can be used to selectively remove surface layers for subsequent microchemical analysis using infrared and tip-enhanced Raman spectroscopy. With its ability to image individual atoms, the AFM is a remarkably sensitive detector that can be used to detect separated components. These diverse functional components of microfluidic manipulation have been combined in this work to demonstrate AFM mediated chromatography. AFM mediated chromatography uses channel-less, shear-driven pumping. This is demonstrated with a thin, aluminum oxide substrate and a non-polar solvent system to separate a mixture of lipophilic dyes. In conventional chromatographic terms, this is analogous to thin-layer chromatography using normal phase alumina substrate with sheardriven pumping provided by the AFM tip-cantilever mechanism. The AFM detection of separated components is accomplished by exploiting the variation in the localized friction of the separated components. The AFM tip-cantilever provides the mechanism for producing shear-induced flows and rapid pumping. Shear-driven chromatography (SDC) is a relatively new concept that overcomes the speed and miniaturization limitations of conventional liquid chromatography. SDC is based on a

  3. Operating microscopes: past, present, and future.

    Science.gov (United States)

    Uluç, Kutluay; Kujoth, Gregory C; Başkaya, Mustafa K

    2009-09-01

    The operating microscope is a fixture of modern surgical facilities, and it is a critically important factor in the success of many of the most complex and difficult surgical interventions used in medicine today. The rise of this key surgical tool reflects advances in understanding the principles of optics and vision that have occurred over centuries. The development of reading spectacles in the late 13th century led to the construction of early compound microscopes in the 16th and 17th centuries by Lippershey, Janssen, Galileo, Hooke, and others. Perhaps surprisingly, Leeuwenhoek's simple microscopes of this era offered improved performance over his contemporaries' designs. The intervening years saw improvements that reduced the spherical and chromatic aberrations present in compound microscopes. By the late 19th century, Carl Zeiss and Ernst Abbe ushered the compound microscope into the beginnings of the modern era of commercial design and production. The introduction of the microscope into the operating room by Nylén in 1921 initiated a revolution in surgical practice that gained momentum throughout the 1950s with multiple refinements, the introduction of the Zeiss OPMI series, and Kurze's application of the microscope to neurosurgery in 1957. Many of the refinements of the last 50 years have greatly improved the handling and practical operation of the surgical microscope, considerations which are equally important to its optical performance. Today's sophisticated operating microscopes allow for advanced real-time angiographic and tumor imaging. In this paper the authors discuss what might be found in the operating rooms of tomorrow.

  4. Comparison between optical techniques and confocal microscopy for defect detection on thin wires

    Energy Technology Data Exchange (ETDEWEB)

    Siegmann, Philip; Sanchez-Brea, Luis Miguel; Martinez-Anton, Juan Carlos; Bernabeu, Eusebio

    2004-11-15

    Conventional microscopy techniques, such as atomic force microscopy (AFM), scanning electron microscopy (SEM), and confocal microscopy (CM) are not suitable for on-line surface inspection of fine metallic wires. In the recent years, some optical techniques have been developed to be used for those tasks. However, they need a rigorous validation. In this work, we have used confocal microscopy to obtain the topography z(x,y) of wires with longitudinal defects, such as dielines. The topography has been used to predict the light scattered by the wire. These simulations have been compared with experimental results, showing a good agreement.

  5. 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

  6. Improved signal model for confocal sensors accounting for object depending artifacts.

    Science.gov (United States)

    Mauch, Florian; Lyda, Wolfram; Gronle, Marc; Osten, Wolfgang

    2012-08-27

    The conventional signal model of confocal sensors is well established and has proven to be exceptionally robust especially when measuring rough surfaces. Its physical derivation however is explicitly based on plane surfaces or point like objects, respectively. Here we show experimental results of a confocal point sensor measurement of a surface standard. The results illustrate the rise of severe artifacts when measuring curved surfaces. On this basis, we present a systematic extension of the conventional signal model that is proven to be capable of qualitatively explaining these artifacts.

  7. Comparison between optical techniques and confocal microscopy for defect detection on thin wires

    International Nuclear Information System (INIS)

    Siegmann, Philip; Sanchez-Brea, Luis Miguel; Martinez-Anton, Juan Carlos; Bernabeu, Eusebio

    2004-01-01

    Conventional microscopy techniques, such as atomic force microscopy (AFM), scanning electron microscopy (SEM), and confocal microscopy (CM) are not suitable for on-line surface inspection of fine metallic wires. In the recent years, some optical techniques have been developed to be used for those tasks. However, they need a rigorous validation. In this work, we have used confocal microscopy to obtain the topography z(x,y) of wires with longitudinal defects, such as dielines. The topography has been used to predict the light scattered by the wire. These simulations have been compared with experimental results, showing a good agreement

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

    Science.gov (United States)

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

    2009-03-02

    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.

  9. The Mathematical Microscope - Making the inaccessible accessible

    DEFF Research Database (Denmark)

    Ottesen, Johnny T.

    2011-01-01

      In this chapter we introduce a new term, the "Mathematical Microscope", as a method of using mathematics in accessing information about reality when this information is otherwise inaccessible. Furthermore, we discuss how models and experiments are related: none of which are important without...... the other. In the sciences and medicine, a link that is often missing in the chain of a system can be made visible with the aid of the mathematical microscope. The mathematical microscope serves not only as a lens to clarify a blurred picture but more important as a tool to unveil profound truths...

  10. Microscopes for NASA's Phoenix Mars Lander

    Science.gov (United States)

    2007-01-01

    One part of the Microscopy, Electrochemistry, and Conductivity Analyzer instrument for NASA's Phoenix Mars Lander is a pair of telescopes with a special wheel (on the right in this photograph) for presenting samples to be inspected with the microscopes. A horizontally mounted optical microscope (on the left in this photograph) and an atomic force microscope will examine soil particles and possibly ice particles. The shapes and the size distributions of soil particles may tell scientists about environmental conditions the material has experienced. Tumbling rounds the edges. Repeated wetting and freezing causes cracking. Clay minerals formed during long exposure to water have distinctive, platy particles shapes.

  11. 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.

  12. phytochemical and microscopical evaluation of desmodium velutinum

    African Journals Online (AJOL)

    USER

    2015-06-01

    , transverse and longitudinal sections of the leaves of .... quantity for preliminary screening and biological studies. Microscopical Examination .... subnitrate dissolved in 20ml acetic acid, made up to 100ml with distilled water).

  13. Feedback regulation of microscopes by image processing.

    Science.gov (United States)

    Tsukada, Yuki; Hashimoto, Koichi

    2013-05-01

    Computational microscope systems are becoming a major part of imaging biological phenomena, and the development of such systems requires the design of automated regulation of microscopes. An important aspect of automated regulation is feedback regulation, which is the focus of this review. As modern microscope systems become more complex, often with many independent components that must work together, computer control is inevitable since the exact orchestration of parameters and timings for these multiple components is critical to acquire proper images. A number of techniques have been developed for biological imaging to accomplish this. Here, we summarize the basics of computational microscopy for the purpose of building automatically regulated microscopes focus on feedback regulation by image processing. These techniques allow high throughput data acquisition while monitoring both short- and long-term dynamic phenomena, which cannot be achieved without an automated system. © 2013 The Authors Development, Growth & Differentiation © 2013 Japanese Society of Developmental Biologists.

  14. Assessing and benchmarking multiphoton microscopes for biologists.

    Science.gov (United States)

    Corbin, Kaitlin; Pinkard, Henry; Peck, Sebastian; Beemiller, Peter; Krummel, Matthew F

    2014-01-01

    Multiphoton microscopy has become staple tool for tracking cells within tissues and organs due to superior depth of penetration, low excitation volumes, and reduced phototoxicity. Many factors, ranging from laser pulse width to relay optics to detectors and electronics, contribute to the overall ability of these microscopes to excite and detect fluorescence deep within tissues. However, we have found that there are few standard ways already described in the literature to distinguish between microscopes or to benchmark existing microscopes to measure the overall quality and efficiency of these instruments. Here, we discuss some simple parameters and methods that can either be used within a multiphoton facility or by a prospective purchaser to benchmark performance. This can both assist in identifying decay in microscope performance and in choosing features of a scope that are suited to experimental needs. © 2014 Elsevier Inc. All rights reserved.

  15. Nucleon relativistic phenomenological and microscopic optical potential

    International Nuclear Information System (INIS)

    Shen Qingbiao; Feng Dachun; Zhuo Yizhong

    1991-01-01

    In this talk, both the phenomenological and microscopic nucleon relativistic optical potentials are presented. The global neutron relativistic phenomenological optical potential (RPOP) based on the available experimental data for various nuclei ranging from C to U with incident energies En=20-1000 MeV has been obtained through automatic search of the best parameters by computer. Then the nucleon relativistic microscopic optical potential (RMOP) is studied by utilizing effective lagrangian based on popular Walecka model. Through comparison between the theoretical results and experimental data we have shed some insight into both the RMOP and RPOP. We have concluded that both the phenomenological and microscopic relativistic optical potentials proposed here can be extensively used for intermediate energy nucleon data evaluation. Further improvement concerning how to combine the phenomenological potential with the microscopic one in order to reduce the number of free parameters appearing in RPOP is suggested. (author). 33 refs, 24 figs

  16. Understanding and caring for an operating microscope

    Directory of Open Access Journals (Sweden)

    Ismael Cordero

    2014-04-01

    Full Text Available An operating or surgical microscope is an optical instrument that provides the surgeon with a stereoscopic, high quality magnified and illuminated image of the small structures in the surgical area.

  17. Assessing and benchmarking multiphoton microscopes for biologists

    Science.gov (United States)

    Corbin, Kaitlin; Pinkard, Henry; Peck, Sebastian; Beemiller, Peter; Krummel, Matthew F.

    2017-01-01

    Multiphoton microscopy has become staple tool for tracking cells within tissues and organs due to superior depth of penetration, low excitation volumes, and reduced phototoxicity. Many factors, ranging from laser pulse width to relay optics to detectors and electronics, contribute to the overall ability of these microscopes to excite and detect fluorescence deep within tissues. However, we have found that there are few standard ways already described in the literature to distinguish between microscopes or to benchmark existing microscopes to measure the overall quality and efficiency of these instruments. Here, we discuss some simple parameters and methods that can either be used within a multiphoton facility or by a prospective purchaser to benchmark performance. This can both assist in identifying decay in microscope performance and in choosing features of a scope that are suited to experimental needs. PMID:24974026

  18. A pragmatic guide to multiphoton microscope design

    Science.gov (United States)

    Young, Michael D.; Field, Jeffrey J.; Sheetz, Kraig E.; Bartels, Randy A.; Squier, Jeff

    2016-01-01

    Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope. PMID:27182429

  19. X ray imaging microscope for cancer research

    Science.gov (United States)

    Hoover, Richard B.; Shealy, David L.; Brinkley, B. R.; Baker, Phillip C.; Barbee, Troy W., Jr.; Walker, Arthur B. C., Jr.

    1991-01-01

    The NASA technology employed during the Stanford MSFC LLNL Rocket X Ray Spectroheliograph flight established that doubly reflecting, normal incidence multilayer optics can be designed, fabricated, and used for high resolution x ray imaging of the Sun. Technology developed as part of the MSFC X Ray Microscope program, showed that high quality, high resolution multilayer x ray imaging microscopes are feasible. Using technology developed at Stanford University and at the DOE Lawrence Livermore National Laboratory (LLNL), Troy W. Barbee, Jr. has fabricated multilayer coatings with near theoretical reflectivities and perfect bandpass matching for a new rocket borne solar observatory, the Multi-Spectral Solar Telescope Array (MSSTA). Advanced Flow Polishing has provided multilayer mirror substrates with sub-angstrom (rms) smoothnesss for the astronomical x ray telescopes and x ray microscopes. The combination of these important technological advancements has paved the way for the development of a Water Window Imaging X Ray Microscope for cancer research.

  20. A lensless, automated microscope for disease diagnostics

    CSIR Research Space (South Africa)

    Hugo, S

    2012-03-01

    Full Text Available using a conventional bright field microscope. Applications of this work are targeted towards the implementation of a full blood count, which could provide resource-limited areas with improved healthcare facilities and diagnosis times....

  1. A lensless, automated microscope for disease diagnostics

    CSIR Research Space (South Africa)

    Hugo, S

    2011-09-01

    Full Text Available microscope. Applications of this work are targeted towards the implementation of a full blood count, which could provide resource-limited areas with improved healthcare facilities and diagnosis times....

  2. Optics of high-performance electron microscopes*

    OpenAIRE

    Rose, H H

    2016-01-01

    During recent years, the theory of charged particle optics together with advances in fabrication tolerances and experimental techniques has lead to very significant advances in high-performance electron microscopes. Here, we will describe which theoretical tools, inventions and designs have driven this development. We cover the basic theory of higher-order electron optics and of image formation in electron microscopes. This leads to a description of different methods to correct aberrations by...

  3. Influence of Confocal Scanning Laser Microscopy specific acquisition parameters on the detection and matching of Speeded-Up Robust Features

    International Nuclear Information System (INIS)

    Stanciu, Stefan G.; Hristu, Radu; Stanciu, George A.

    2011-01-01

    The robustness and distinctiveness of local features to various object or scene deformations and to modifications of the acquisition parameters play key roles in the design of many computer vision applications. In this paper we present the results of our experiments on the behavior of a recently developed technique for local feature detection and description, Speeded-Up Robust Features (SURF), regarding image modifications specific to Confocal Scanning Laser Microscopy (CSLM). We analyze the repeatability of detected SURF keypoints and the precision-recall of their matching under modifications of three important CSLM parameters: pinhole aperture, photomultiplier (PMT) gain and laser beam power. During any investigation by CSLM these three parameters have to be modified, individually or together, in order to optimize the contrast and the Signal Noise Ratio (SNR), being also inherently modified when changing the microscope objective. Our experiments show that an important amount of SURF features can be detected at the same physical locations in images collected at different values of the pinhole aperture, PMT gain and laser beam power, and further on can be successfully matched based on their descriptors. In the final part, we exemplify the potential of SURF in CSLM imaging by presenting a SURF-based computer vision application that deals with the mosaicing of images collected by this technique. -- Research highlights: → Influence of pinhole aperture modifications on SURF detection and matching in CSLM images. → Influence of photomultiplier gain modifications on SURF detection and matching in CSLM images. → Influence of laser beam power modifications on SURF detection and matching in CSLM images. → SURF-based automated mosaicing of CSLM images.

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

    Directory of Open Access Journals (Sweden)

    Meiyan Li

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

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

    DEFF Research Database (Denmark)

    Schiek, Manuela; Balzer, Frank

    2014-01-01

    . Control of these interfaces directly impacts on the performance and here we show with basic growth studies of model compounds on dielectric and graphitic surfaces, the formation of distinctly textured films. Silver-nanowire meshes are presented as an alternative transparent electrode material. Confocal...

  6. Confocal laser induced fluorescence with comparable spatial localization to the conventional method

    Science.gov (United States)

    Thompson, Derek S.; Henriquez, Miguel F.; Scime, Earl E.; Good, Timothy N.

    2017-10-01

    We present measurements of ion velocity distributions obtained by laser induced fluorescence (LIF) using a single viewport in an argon plasma. A patent pending design, which we refer to as the confocal fluorescence telescope, combines large objective lenses with a large central obscuration and a spatial filter to achieve high spatial localization along the laser injection direction. Models of the injection and collection optics of the two assemblies are used to provide a theoretical estimate of the spatial localization of the confocal arrangement, which is taken to be the full width at half maximum of the spatial optical response. The new design achieves approximately 1.4 mm localization at a focal length of 148.7 mm, improving on previously published designs by an order of magnitude and approaching the localization achieved by the conventional method. The confocal method, however, does so without requiring a pair of separated, perpendicular optical paths. The confocal technique therefore eases the two window access requirement of the conventional method, extending the application of LIF to experiments where conventional LIF measurements have been impossible or difficult, or where multiple viewports are scarce.

  7. Resolution enhancement by subtraction of confocal signals taken at different pinhole sizes.

    Science.gov (United States)

    Heintzmann, Rainer; Sarafis, Vassilios; Munroe, Paul; Nailon, John; Hanley, Quentin S; Jovin, Thomas M

    2003-01-01

    Subtractive imaging in confocal fluorescence light microscopy is based on the subtraction of a suitably weighted widefield image from a confocal image. An approximation to a widefield image can be obtained by detection with an opened confocal pinhole. The subtraction of images enhances the resolution in-plane as well as along the optic axis. Due to the linearity of the approach, the effect of subtractive imaging in Fourier-space corresponds to a reduction of low spatial frequency contributions leading to a relative enhancement of the high frequencies. Along the direction of the optic axis this also results in an improved sectioning. Image processing can achieve a similar effect. However, a 3D volume dataset must be acquired and processed, yielding a result essentially identical to subtractive imaging but superior in signal-to-noise ratio. The latter can be increased further with the technique of weighted averaging in Fourier-space. A comparison of 2D and 3D experimental data analysed with subtractive imaging, the equivalent Fourier-space processing of the confocal data only, and Fourier-space weighted averaging is presented.

  8. Confocal microscopy of chloroplast morphology and ontogeny in three strains of Dictyochloropsis (Trebouxiophyceae, Chlorophyta)

    Czech Academy of Sciences Publication Activity Database

    Škaloud, P.; Neustupa, J.; Radochová, Barbora; Kubínová, Lucie

    2005-01-01

    Roč. 44, č. 3 (2005), s. 261-269 ISSN 0031-8884 Grant - others:FRVŠ(CZ) 2826/2003; IGA MŠk(CZ) 139/2002/B Institutional research plan: CEZ:AV0Z5011922 Keywords : chloroplast ontogeny * confocal microscopy * Dichtyochloropsis Subject RIV: EF - Botanics Impact factor: 1.271, year: 2005

  9. A novel method for evaluation of capillarity in human skeletal muscles from confocal 3D images

    Czech Academy of Sciences Publication Activity Database

    Janáček, Jiří; Cvetko, E.; Kubínová, Lucie; Travnik, L.; Eržen, I.

    2011-01-01

    Roč. 81, č. 2 (2011), s. 231-238 ISSN 0026-2862 R&D Projects: GA MŠk(CZ) MEB090910; GA MŠk(CZ) LC06063 Institutional research plan: CEZ:AV0Z50110509 Keywords : confocal microscopy * 3D * capillaries * skeletal muscle * human Subject RIV: BO - Biophysics Impact factor: 2.828, year: 2011

  10. Design and Performance of a Multi-Point Scan Confocal Microendoscope

    Directory of Open Access Journals (Sweden)

    Matthew D. Risi

    2014-11-01

    Full Text Available Confocal fluorescence microendoscopy provides high-resolution cellular-level imaging via a minimally invasive procedure, but requires fast scanning to achieve real-time imaging in vivo. Ideal confocal imaging performance is obtained with a point scanning system, but the scan rates required for in vivo biomedical imaging can be difficult to achieve. By scanning a line of illumination in one direction in conjunction with a stationary confocal slit aperture, very high image acquisition speeds can be achieved, but at the cost of a reduction in image quality. Here, the design, implementation, and experimental verification of a custom multi-point aperture modification to a line-scanning multi-spectral confocal microendoscope is presented. This new design improves the axial resolution of a line-scan system while maintaining high imaging rates. In addition, compared to the line-scanning configuration, previously reported simulations predicted that the multi-point aperture geometry greatly reduces the effects of tissue scatter on image quality. Experimental results confirming this prediction are presented.

  11. Real-time evaluation of aggregation using confocal imaging and image analysis tools.

    Science.gov (United States)

    Hamrang, Zahra; Zindy, Egor; Clarke, David; Pluen, Alain

    2014-02-07

    Real-time confocal imaging was utilised to monitor the in situ loss of BSA monomers and aggregate formation using Spatial Intensity Distribution Analysis (SpIDA) and Raster Image Correlation Spectroscopy (RICS). At the proof of concept level this work has demonstrated the applicability of RICS and SpIDA for monitoring protein oligomerisation and larger aggregate formation.

  12. Diagnostic accuracy of confocal microscopy imaging vs. punch biopsy for diagnosing and subtyping basal cell carcinoma

    NARCIS (Netherlands)

    Kadouch, D. J.; Leeflang, M. M.; Elshot, Y. S.; Longo, C.; Ulrich, M.; van der Wal, A. C.; Wolkerstorfer, A.; Bekkenk, M. W.; de Rie, M. A.

    2017-01-01

    BackgroundIn vivo reflectance confocal microscopy (RCM) is a promising non-invasive skin imaging technique that could facilitate early diagnosis of basal cell carcinoma (BCC) instead of routine punch biopsies. However, the clinical value and utility of RCM vs. a punch biopsy in diagnosing and

  13. Reflectance confocal microscopy: an effective tool for monitoring ultraviolet B phototherapy in psoriasis.

    NARCIS (Netherlands)

    Wolberink, E.A.W.; Erp, P.E.J. van; Boer-van Huizen, R.T. de; Kerkhof, P.C.M. van de; Gerritsen, M.J.P.

    2012-01-01

    Background In vivo reflectance confocal microscopy (RCM) is a novel, noninvasive imaging technique which enables imaging of skin at a cellular resolution comparable to conventional microscopy. Objectives We performed a pilot study to evaluate RCM as a noninvasive tool for monitoring ultraviolet (UV)

  14. Cellular features of psoriatic skin: imaging and quantification using in vivo reflectance confocal microscopy

    NARCIS (Netherlands)

    Wolberink, E.A.W.; Erp, P.E.J. van; Teussink, M.M.; Kerkhof, P.C.M. van de; Gerritsen, M.J.P.

    2011-01-01

    BACKGROUND: In vivo reflectance confocal microscopy (RCM) is a novel, exciting imaging technique. It provides images of cell-and tissue structures and dynamics in situ, in real time, without the need for ex vivo tissue samples. RCM visualizes the superficial part of human skin up to a depth of 250

  15. Correcting the axial shrinkage of skeletal muscle thick sections visualized by confocal microscopy

    Czech Academy of Sciences Publication Activity Database

    Janáček, Jiří; Kreft, M.; Čebašek, V.; Eržen, I.

    2012-01-01

    Roč. 246, č. 2 (2012), s. 107-112 ISSN 0022-2720 R&D Projects: GA MŠk(CZ) MEB090910; GA MŠk(CZ) LC06063 Institutional research plan: CEZ:AV0Z50110509 Keywords : capillaries * confocal microscopy * sample deformation * shrinkage * skeletal muscle * 3D Subject RIV: FH - Neurology Impact factor: 1.633, year: 2012

  16. Double-confocal resonator for X-ray generation via intracavity Thomson scattering

    Energy Technology Data Exchange (ETDEWEB)

    Xie, M. [Lawrence Berkeley Lab., CA (United States)

    1995-12-31

    There has been a growing interest in developing compact X-ray sources through Thomson scattering of a laser beam by a relativistic electron beam. For higher X-ray flux it is desirable to have the scattering to occur inside an optical resonator where the laser power is higher. In this paper I propose a double-confocal resonator design optimized for head-on Thomson scattering inside an FEL oscillator and analyze its performance taking into account the diffraction and FEL gain. A double confocal resonator is equivalent to two confocal resonators in series. Such a resonator has several advantages: it couples electron beam through and X-ray out of the cavity with holes on cavity mirrors, thus allowing the system to be compact; it supports the FEL mode with minimal diffraction loss through the holes; it provides a laser focus in the forward direction for a better mode overlap with the electron beam; and it provides a focus at the same location in the backward direction for higher Thomson scattering efficiency; in addition, the mode size at the focal point and hence the Rayleigh range can be adjusted simply through intracavity apertures; furthermore, it gives a large mode size at the mirrors to reduce power loading. Simulations as well as analytical results will be presented. Also other configurations of intracavity Thomson scattering where the double-confocal resonator could be useful will be discussed.

  17. [Contribution of confocal microscopy and anterior chamber OCT to the study of corneal endothelial pathologies].

    Science.gov (United States)

    Fayol, N; Labbé, A; Dupont-Monod, S; Dupas, B; Baudouin, C

    2007-04-01

    To describe the appearance of various endothelial diseases with in vivo confocal microscopy and anterior chamber optical coherence tomography (AC OCT). In this study, ten patients with five different corneal endothelial pathologies were evaluated. Three patients had cornea guttata, three had corneal endothelial precipitates, two had irido-corneo-endothelial (ICE) syndrome, one had endothelial folds, and one had breaks in the Descemet membrane. All patients had bilateral ophthalmologic examinations, in vivo confocal microscopy, and AC OCT analysis. In cases of cornea guttata, AC OCT showed a finely embossed line corresponding to the empty intercellular cavities found with in vivo confocal microscopy. Corneal endothelium precipitates had the aspect of round formations suspended with the endothelium. Iris atrophy and irido-corneal synechiae resulting from ICE syndrome were precisely visualized with the AC OCT. High-resolution images of the anterior segment could be obtained using the AC OCT. Associated with in vivo confocal microscopy, these two new imaging techniques provide a precise evaluation of endothelial pathologies.

  18. [In vivo confocal microscopy: a new tool for the diagnosis of Acanthamoeba keratitis].

    Science.gov (United States)

    Grise-Dulac, A; Brasnu, E; Goldchmidt, P; Dupas, B; Labbe, A; Borderie, V; Borsali, E; Chaumeil, C; Baudouin, C

    2010-06-01

    To study the usefulness of in vivo confocal microscopy imaging for the diagnosis of Acanthamoeba keratitis. A retrospective review of 50 cases of Acanthamoeba keratitis followed at the Quinze-Vingts National Ophthalmology Hospital from January 2005 to July 2008 was conducted. Gender, age, contact lens wear, best-corrected visual acuity before and after treatment, slit-lamp examination findings, corneal scrapings for biological analysis, and in vivo confocal microscopy images were analyzed. Nearly 82% of the cases of keratitis had a history of contact lens wear. Polymerase chain reaction (PCR) was positive for 40% of the samples. Heidelberg Retinal Tomograph II-Rostock Cornea Module (HRTII-RCM) examination detected images evoking Acanthamoeba cyst-like images in 84% of the cases. When the quality of biological samples was inadequate, the assessment of Acanthamoeba cysts using in vivo confocal microscopy made it possible to orient the diagnosis and to partially explain favorable progression under treatment. This technique showed images suggesting combined Acanthamoeba and fungal keratitis. HRTII-RCM in vivo confocal microscopy is a non invasive and rapid technique that may be helpful for the diagnosis of Acanthamoeba keratitis, especially when laboratory testing is not contributive and when Acanthamoeba keratitis is combined with a fungal infection. Copyright (c) 2010 Elsevier Masson SAS. All rights reserved.

  19. Various confocal scan features of cysts and trophozoites in cases with Acanthamoeba keratitis.

    Science.gov (United States)

    Rezaei Kanavi, Mozhgan; Naghshgar, Nima; Javadi, Mohammad Ali; Sadat Hashemi, Marzieh

    2012-01-01

    To describe the various confocal scan features of cysts and trophozoites in patients with Acanthamoeba keratitis and to specify the associated findings. In a retrospective study of cases between June 2005 and June 2010, we reviewed all the recorded confocal scan images of patients given a high index in regards to clinical suspicion of Acanthamoeba keratitis, in order to specify the various morphometric and morphologic features of Acanthamoeba cysts and trophozoites and to characterize the associated findings in such cases. Confocal scan images of 170 eyes from 170 patients were reviewed. Bilayered, target-shaped, coffee-bean and rod-shaped appearances of the cysts were observed in 100%, 82.9%, 36.4%, and 17.5% of cases, respectively. Single file arrangement of the cysts was noticed in 22 cases. The mean size of the cysts was 18.9 µm (range 10-39.6). In all cases, trophozoites were observed as pear-shaped or irregularly wedge-shaped structures, some surrounded by a brilliant halo and some exhibiting fine pseudopodia-like extensions, with mean size of 30.2 µm (range 19.2-55.6). Keratoneuritis and the anterior stromal honeycomb pattern were seen in 28.2% and 5.9% of cases, respectively. To our knowledge, this is the largest case-series study on confocal scan features of Acanthamoeba cysts and trophozoites in cases with clinical diagnosis of Acanthamoeba keratitis specifying the morphologic and morphometric criteria of this infectious organism and the associated findings.

  20. In vivo Diagnosis of Basal Cell Carcinoma Subtype by Reflectance Confocal Microscopy

    NARCIS (Netherlands)

    Peppelman, M.; Wolberink, E.A.W.; Blokx, W.A.M.; Kerkhof, P.C.M. van de; Erp, P.E.J. van; Gerritsen, M.J.P.

    2013-01-01

    Background: Reflectance confocal microscopy (RCM) is a noninvasive imaging technique. Currently, RCM is mainly used for the diagnosis of melanoma and nonmelanoma skin cancer including basal cell carcinoma (BCC). Until now, it has not been possible to distinguish between subtypes of BCC using RCM.