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Sample records for 3d optical microscopy

  1. Deconvolution in 3-D optical microscopy.

    PubMed

    Shaw, P

    1994-09-01

    Fluorescent probes are becoming ever more widely used in the study of subcellular structure, and determination of their three-dimensional distributions has become very important. Confocal microscopy is now a common technique for overcoming the problem of out-of-focus flare in fluorescence imaging, but an alternative method uses digital image processing of conventional fluorescence images--a technique often termed 'deconvolution' or 'restoration'. This review attempts to explain image deconvolution in a non-technical manner. It is also applicable to 3-D confocal images, and can provide a further significant improvement in clarity and interpretability of such images. Some examples of the application of image deconvolution to both conventional and confocal fluorescence images are shown.

  2. Precision 3-D microscopy with intensity modulated fibre optic scanners

    NASA Astrophysics Data System (ADS)

    Olmos, P.

    2016-01-01

    Optical 3-D imagers constitute a family of precision and useful instruments, easily available on the market in a wide variety of configurations and performances. However, besides their cost they usually provide an image of the object (i.e. a more or less faithful representation of the reality) instead of a truly object's reconstruction. Depending on the detailed working principles of the equipment, this reconstruction may become a challenging task. Here a very simple yet reliable device is described; it is able to form images of opaque objects by illuminating them with an optical fibre and collecting the reflected light with another fibre. Its 3-D capability comes from the spatial filtering imposed by the fibres together with their movement (scanning) along the three directions: transversal (surface) and vertical. This unsophisticated approach allows one to model accurately the entire optical process and to perform the desired reconstruction, finding that information about the surface which is of interest: its profile and its reflectance, ultimately related to the type of material.

  3. Advanced 3D Optical Microscopy in ENS Research.

    PubMed

    Vanden Berghe, Pieter

    2016-01-01

    Microscopic techniques are among the few approaches that have survived the test of time. Being invented half way the seventeenth century by Antonie van Leeuwenhoek and Robert Hooke, this technology is still essential in modern biomedical labs. Many microscopy techniques have been used in ENS research to guide researchers in their dissections and later to enable electrode recordings. Apart from this, microscopy has been instrumental in the identification of subpopulations of cells in the ENS, using a variety of staining methods. A significant step forward in the use of microscopy was the introduction of fluorescence approaches. Due to the fact that intense excitation light is now filtered away from the longer wavelength emission light, the contrast can be improved drastically, which helped to identify subpopulations of enteric neurons in a variety of species. Later functionalized fluorescent probes were used to measure and film activity in muscle and neuronal cells. Another important impetus to the use of microscopy was the discovery and isolation of the green fluorescent protein (GFP), as it gave rise to the development of many different color variants and functionalized constructs. Recent advances in microscopy are the result of a continuous search to enhance contrast between the item of interest and its background but also to improve resolving power to tell two small objects apart. In this chapter three different microscopy approaches will be discussed that can aid to improve our understanding of ENS function within the gut wall. PMID:27379646

  4. Advanced 3D Optical Microscopy in ENS Research.

    PubMed

    Vanden Berghe, Pieter

    2016-01-01

    Microscopic techniques are among the few approaches that have survived the test of time. Being invented half way the seventeenth century by Antonie van Leeuwenhoek and Robert Hooke, this technology is still essential in modern biomedical labs. Many microscopy techniques have been used in ENS research to guide researchers in their dissections and later to enable electrode recordings. Apart from this, microscopy has been instrumental in the identification of subpopulations of cells in the ENS, using a variety of staining methods. A significant step forward in the use of microscopy was the introduction of fluorescence approaches. Due to the fact that intense excitation light is now filtered away from the longer wavelength emission light, the contrast can be improved drastically, which helped to identify subpopulations of enteric neurons in a variety of species. Later functionalized fluorescent probes were used to measure and film activity in muscle and neuronal cells. Another important impetus to the use of microscopy was the discovery and isolation of the green fluorescent protein (GFP), as it gave rise to the development of many different color variants and functionalized constructs. Recent advances in microscopy are the result of a continuous search to enhance contrast between the item of interest and its background but also to improve resolving power to tell two small objects apart. In this chapter three different microscopy approaches will be discussed that can aid to improve our understanding of ENS function within the gut wall.

  5. Dynamic complex optical fields for optical manipulation, 3D microscopy, and photostimulation of neurotransmitters

    NASA Astrophysics Data System (ADS)

    Daria, Vincent R.; Stricker, Christian; Bekkers, John; Redman, Steve; Bachor, Hans

    2010-08-01

    We demonstrate a multi-functional system capable of multiple-site two-photon excitation of photo-sensitive compounds as well as transfer of optical mechanical properties on an array of mesoscopic particles. We use holographic projection of a single Ti:Sapphire laser operating in femtosecond pulse mode to show that the projected three-dimensional light patterns have sufficient spatiotemporal photon density for multi-site two-photon excitation of biological fluorescent markers and caged neurotransmitters. Using the same laser operating in continuous-wave mode, we can use the same light patterns for non-invasive transfer of both linear and orbital angular momentum on a variety of mesoscopic particles. The system also incorporates high-speed scanning using acousto-optic modulators to rapidly render 3D images of neuron samples via two-photon microscopy.

  6. Towards non-invasive 3D hepatotoxicity assays with optical coherence phase microscopy

    NASA Astrophysics Data System (ADS)

    Nelson, Leonard J.; Koulovasilopoulos, Andreas; Treskes, Philipp; Hayes, Peter C.; Plevris, John N.; Bagnaninchi, Pierre O.

    2015-03-01

    Three-dimensional tissue-engineered models are increasingly recognised as more physiologically-relevant than standard 2D cell culture for pre-clinical drug toxicity testing. However, many types of conventional toxicity assays are incompatible with dense 3D tissues. This study investigated the use of optical coherence phase microscopy (OCPM) as a novel approach to assess cell death in 3D tissue culture. For 3D micro-spheroid formation Human hepatic C3A cells were encapsulated in hyaluronic acid gels and cultured in 100μl MEME/10%FBS in 96-well plates. After spheroid formation the 3D liver constructs were exposed to acetaminophen on culture day 8. Acetaminophen hepatotoxicity in 3D cultures was evaluated using standard biochemical assays. An inverted OCPM in common path configuration was developed with a Callisto OCT engine (Thorlabs), centred at 930nm and a custom scanning head. Intensity data were used to perform in-depth microstructural imaging. In addition, phase fluctuations were measured by collecting several successive B scans at the same location, and statistics on the first time derivative of the phase, i.e. time fluctuations, were analysed over the acquisition time interval to retrieve overall cell viability. OCPM intensity (cell cluster size) and phase fluctuation statistics were directly compared with biochemical assays. In this study, we investigated optical coherence phase tomography to assess cell death in a 3d liver model after exposure to a prototypical hepatotoxin, acetaminophen. We showed that OCPM has the potential to assess noninvasively and label-free drug toxicity in 3D tissue models.

  7. Computational-optical microscopy for 3D biological imaging beyond the diffraction limit

    NASA Astrophysics Data System (ADS)

    Grover, Ginni

    In recent years, super-resolution imaging has become an important fluorescent microscopy tool. It has enabled imaging of structures smaller than the optical diffraction limit with resolution less than 50 nm. Extension to high-resolution volume imaging has been achieved by integration with various optical techniques. In this thesis, development of a fluorescent microscope to enable high resolution, extended depth, three dimensional (3D) imaging is discussed; which is achieved by integration of computational methods with optical systems. In the first part of the thesis, point spread function (PSF) engineering for volume imaging is discussed. A class of PSFs, referred to as double-helix (DH) PSFs, is generated. The PSFs exhibit two focused spots in the image plane which rotate about the optical axis, encoding depth in rotation of the image. These PSFs extend the depth-of-field up to a factor of ˜5. Precision performance of the DH-PSFs, based on an information theoretical analysis, is compared with other 3D methods with conclusion that the DH-PSFs provide the best precision and the longest depth-of-field. Out of various possible DH-PSFs, a suitable PSF is obtained for super-resolution microscopy. The DH-PSFs are implemented in imaging systems, such as a microscope, with a special phase modulation at the pupil plane. Surface-relief elements which are polarization-insensitive and ˜90% light efficient are developed for phase modulation. The photon-efficient DH-PSF microscopes thus developed are used, along with optimal position estimation algorithms, for tracking and super-resolution imaging in 3D. Imaging at depths-of-field of up to 2.5 microm is achieved without focus scanning. Microtubules were imaged with 3D resolution of (6, 9, 39) nm, which is in close agreement with the theoretical limit. A quantitative study of co-localization of two proteins in volume was conducted in live bacteria. In the last part of the thesis practical aspects of the DH-PSF microscope are

  8. Computational optical-sectioning microscopy for 3D quantization of cell motion: results and challenges

    NASA Astrophysics Data System (ADS)

    McNally, James G.

    1994-09-01

    How cells move and navigate within a 3D tissue mass is of central importance in such diverse problems as embryonic development, wound healing and metastasis. This locomotion can now be visualized and quantified by using computation optical-sectioning microscopy. In this approach, a series of 2D images at different depths in a specimen are stacked to construct a 3D image, and then with a knowledge of the microscope's point-spread function, the actual distribution of fluorescent intensity in the specimen is estimated via computation. When coupled with wide-field optics and a cooled CCD camera, this approach permits non-destructive 3D imaging of living specimens over long time periods. With these techniques, we have observed a complex diversity of motile behaviors in a model embryonic system, the cellular slime mold Dictyostelium. To understand the mechanisms which control these various behaviors, we are examining motion in various Dictyostelium mutants with known defects in proteins thought to be essential for signal reception, cell-cell adhesion or locomotion. This application of computational techniques to analyze 3D cell locomotion raises several technical challenges. Image restoration techniques must be fast enough to process numerous 1 Gbyte time-lapse data sets (16 Mbytes per 3D image X 60 time points). Because some cells are weakly labeled and background intensity is often high due to unincorporated dye, the SNR in some of these images is poor. Currently, the images are processed by a regularized linear least- squares restoration method, and occasionally by a maximum-likelihood method. Also required for these studies are accurate automated- tracking procedures to generate both 3D trajectories for individual cells and 3D flows for a group of cells. Tracking is currently done independently for each cell, using a cell's image as a template to search for a similar image at the next time point. Finally, sophisticated visualization techniques are needed to view the

  9. In situ 3D characterization of historical coatings and wood using multimodal nonlinear optical microscopy.

    PubMed

    Latour, Gaël; Echard, Jean-Philippe; Didier, Marie; Schanne-Klein, Marie-Claire

    2012-10-22

    We demonstrate multimodal nonlinear optical imaging of historical artifacts by combining Second Harmonic Generation (SHG) and Two-Photon Excited Fluorescence (2PEF) microscopies. We first identify the nonlinear optical response of materials commonly encountered in coatings of cultural heritage artifacts by analyzing one- and multi-layered model samples. We observe 2PEF signals from cochineal lake and sandarac and show that pigments and varnish films can be discriminated by exploiting their different emission spectral ranges as in luminescence linear spectroscopy. We then demonstrate SHG imaging of a filler, plaster, composed of bassanite particles which exhibit a non centrosymmetric crystal structure. We also show that SHG/2PEF imaging enables the visualization of wood microstructure through typically 60 µm-thick coatings by revealing crystalline cellulose (SHG signal) and lignin (2PEF signal) in the wood cell walls. Finally, in situ multimodal nonlinear imaging is demonstrated in a historical violin. SHG/2PEF imaging thus appears as a promising non-destructive and contactless tool for in situ 3D investigation of historical coatings and more generally for wood characterization and coating analysis at micrometer scale. PMID:23187225

  10. Real Time Gabor-Domain Optical Coherence Microscopy for 3D Imaging.

    PubMed

    Rolland, Jannick P; Canavesi, Cristina; Tankam, Patrice; Cogliati, Andrea; Lanis, Mara; Santhanam, Anand P

    2016-01-01

    Fast, robust, nondestructive 3D imaging is needed for the characterization of microscopic tissue structures across various clinical applications. A custom microelectromechanical system (MEMS)-based 2D scanner was developed to achieve, together with a multi-level GPU architecture, 55 kHz fast-axis A-scan acquisition in a Gabor-domain optical coherence microscopy (GD-OCM) custom instrument. GD-OCM yields high-definition micrometer-class volumetric images. A dynamic depth of focusing capability through a bio-inspired liquid lens-based microscope design, as in whales' eyes, was developed to enable the high definition instrument throughout a large field of view of 1 mm3 volume of imaging. Developing this technology is prime to enable integration within the workflow of clinical environments. Imaging at an invariant resolution of 2 μm has been achieved throughout a volume of 1 × 1 × 0.6 mm3, acquired in less than 2 minutes. Volumetric scans of human skin in vivo and an excised human cornea are presented. PMID:27046601

  11. Analytic 3D imaging of mammalian nucleus at nanoscale using coherent x-rays and optical fluorescence microscopy.

    PubMed

    Song, Changyong; Takagi, Masatoshi; Park, Jaehyun; Xu, Rui; Gallagher-Jones, Marcus; Imamoto, Naoko; Ishikawa, Tetsuya

    2014-09-01

    Despite the notable progress that has been made with nano-bio imaging probes, quantitative nanoscale imaging of multistructured specimens such as mammalian cells remains challenging due to their inherent structural complexity. Here, we successfully performed three-dimensional (3D) imaging of mammalian nuclei by combining coherent x-ray diffraction microscopy, explicitly visualizing nuclear substructures at several tens of nanometer resolution, and optical fluorescence microscopy, cross confirming the substructures with immunostaining. This demonstrates the successful application of coherent x-rays to obtain the 3D ultrastructure of mammalian nuclei and establishes a solid route to nanoscale imaging of complex specimens.

  12. Feasibility study on 3-D shape analysis of high-aspect-ratio features using through-focus scanning optical microscopy

    PubMed Central

    Attota, Ravi Kiran; Weck, Peter; Kramar, John A.; Bunday, Benjamin; Vartanian, Victor

    2016-01-01

    In-line metrologies currently used in the semiconductor industry are being challenged by the aggressive pace of device scaling and the adoption of novel device architectures. Metrology and process control of three-dimensional (3-D) high-aspect-ratio (HAR) features are becoming increasingly important and also challenging. In this paper we present a feasibility study of through-focus scanning optical microscopy (TSOM) for 3-D shape analysis of HAR features. TSOM makes use of 3-D optical data collected using a conventional optical microscope for 3-D shape analysis. Simulation results of trenches and holes down to the 11 nm node are presented. The ability of TSOM to analyze an array of HAR features or a single isolated HAR feature is also presented. This allows for the use of targets with area over 100 times smaller than that of conventional gratings, saving valuable real estate on the wafers. Indications are that the sensitivity of TSOM may match or exceed the International Technology Roadmap for Semiconductors (ITRS) measurement requirements for the next several years. Both simulations and preliminary experimental results are presented. The simplicity, lowcost, high throughput, and nanometer scale 3-D shape sensitivity of TSOM make it an attractive inspection and process monitoring solution for nanomanufacturing. PMID:27464112

  13. Feasibility study on 3-D shape analysis of high-aspect-ratio features using through-focus scanning optical microscopy.

    PubMed

    Attota, Ravi Kiran; Weck, Peter; Kramar, John A; Bunday, Benjamin; Vartanian, Victor

    2016-07-25

    In-line metrologies currently used in the semiconductor industry are being challenged by the aggressive pace of device scaling and the adoption of novel device architectures. Metrology and process control of three-dimensional (3-D) high-aspect-ratio (HAR) features are becoming increasingly important and also challenging. In this paper we present a feasibility study of through-focus scanning optical microscopy (TSOM) for 3-D shape analysis of HAR features. TSOM makes use of 3-D optical data collected using a conventional optical microscope for 3-D shape analysis. Simulation results of trenches and holes down to the 11 nm node are presented. The ability of TSOM to analyze an array of HAR features or a single isolated HAR feature is also presented. This allows for the use of targets with area over 100 times smaller than that of conventional gratings, saving valuable real estate on the wafers. Indications are that the sensitivity of TSOM may match or exceed the International Technology Roadmap for Semiconductors (ITRS) measurement requirements for the next several years. Both simulations and preliminary experimental results are presented. The simplicity, lowcost, high throughput, and nanometer scale 3-D shape sensitivity of TSOM make it an attractive inspection and process monitoring solution for nanomanufacturing. PMID:27464112

  14. 3D multiplexed immunoplasmonics microscopy

    NASA Astrophysics Data System (ADS)

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

    2016-07-01

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

  15. 3D multiplexed immunoplasmonics microscopy

    NASA Astrophysics Data System (ADS)

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

    2016-07-01

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

  16. 3D multiplexed immunoplasmonics microscopy.

    PubMed

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

    2016-07-21

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

  17. BigNeuron: Large-scale 3D Neuron Reconstruction from Optical Microscopy Images

    PubMed Central

    Peng, Hanchuan; Hawrylycz, Michael; Roskams, Jane; Hill, Sean; Spruston, Nelson; Meijering, Erik; Ascoli, Giorgio A.

    2016-01-01

    Understanding the structure of single neurons is critical for understanding how they function within neural circuits. BigNeuron is a new community effort that combines modern bioimaging informatics, recent leaps in labeling and microscopy, and the widely recognized need for openness and standardization to provide a community resource for automated reconstruction of dendritic and axonal morphology of single neurons. PMID:26182412

  18. BigNeuron: Large-Scale 3D Neuron Reconstruction from Optical Microscopy Images.

    PubMed

    Peng, Hanchuan; Hawrylycz, Michael; Roskams, Jane; Hill, Sean; Spruston, Nelson; Meijering, Erik; Ascoli, Giorgio A

    2015-07-15

    Understanding the structure of single neurons is critical for understanding how they function within neural circuits. BigNeuron is a new community effort that combines modern bioimaging informatics, recent leaps in labeling and microscopy, and the widely recognized need for openness and standardization to provide a community resource for automated reconstruction of dendritic and axonal morphology of single neurons.

  19. Fast spatial beam shaping by acousto-optic diffraction for 3D non-linear microscopy.

    PubMed

    Akemann, Walther; Léger, Jean-François; Ventalon, Cathie; Mathieu, Benjamin; Dieudonné, Stéphane; Bourdieu, Laurent

    2015-11-01

    Acousto-optic deflection (AOD) devices offer unprecedented fast control of the entire spatial structure of light beams, most notably their phase. AOD light modulation of ultra-short laser pulses, however, is not straightforward to implement because of intrinsic chromatic dispersion and non-stationarity of acousto-optic diffraction. While schemes exist to compensate chromatic dispersion, non-stationarity remains an obstacle. In this work we demonstrate an efficient AOD light modulator for stable phase modulation using time-locked generation of frequency-modulated acoustic waves at the full repetition rate of a high power laser pulse amplifier of 80 kHz. We establish the non-local relationship between the optical phase and the generating acoustic frequency function and verify the system for temporal stability, phase accuracy and generation of non-linear two-dimensional phase functions. PMID:26561090

  20. Fast spatial beam shaping by acousto-optic diffraction for 3D non-linear microscopy.

    PubMed

    Akemann, Walther; Léger, Jean-François; Ventalon, Cathie; Mathieu, Benjamin; Dieudonné, Stéphane; Bourdieu, Laurent

    2015-11-01

    Acousto-optic deflection (AOD) devices offer unprecedented fast control of the entire spatial structure of light beams, most notably their phase. AOD light modulation of ultra-short laser pulses, however, is not straightforward to implement because of intrinsic chromatic dispersion and non-stationarity of acousto-optic diffraction. While schemes exist to compensate chromatic dispersion, non-stationarity remains an obstacle. In this work we demonstrate an efficient AOD light modulator for stable phase modulation using time-locked generation of frequency-modulated acoustic waves at the full repetition rate of a high power laser pulse amplifier of 80 kHz. We establish the non-local relationship between the optical phase and the generating acoustic frequency function and verify the system for temporal stability, phase accuracy and generation of non-linear two-dimensional phase functions.

  1. Optical sectioning and 3D reconstructions as an alternative to scanning electron microscopy for analysis of cell shape1

    PubMed Central

    Landis, Jacob B.; Ventura, Kayla L.; Soltis, Douglas E.; Soltis, Pamela S.; Oppenheimer, David G.

    2015-01-01

    Premise of the study: Visualizing flower epidermal cells is often desirable for investigating the interaction between flowers and their pollinators, in addition to the broader range of ecological interactions in which flowers are involved. We developed a protocol for visualizing petal epidermal cells without the limitations of the commonly used method of scanning electron microscopy (SEM). Methods: Flower material was collected and fixed in glutaraldehyde, followed by dehydration in an ethanol series. Flowers were dissected to collect petals, and subjected to a Histo-Clear series to remove the cuticle. Material was then stained with aniline blue, mounted on microscope slides, and imaged using a compound fluorescence microscope to obtain optical sections that were reconstructed into a 3D image. Results: This optical sectioning method yielded high-quality images of the petal epidermal cells with virtually no damage to cells. Flowers were processed in larger batches than are possible using common SEM methods. Also, flower size was not a limiting factor as often observed in SEM studies. Flowers up to 5 cm in length were processed and mounted for visualization. Conclusions: This method requires no special equipment for sample preparation prior to imaging and should be seen as an alternative method to SEM. PMID:25909040

  2. Gabor-domain optical coherence microscopy with integrated dual-axis MEMS scanner for fast 3D imaging and metrology

    NASA Astrophysics Data System (ADS)

    Canavesi, Cristina; Cogliati, Andrea; Hayes, Adam; Santhanam, Anand P.; Tankam, Patrice; Rolland, Jannick P.

    2015-10-01

    Fast, robust, nondestructive 3D imaging is needed for characterization of microscopic structures in industrial and clinical applications. A custom micro-electromechanical system (MEMS)-based 2D scanner system was developed to achieve 55 kHz A-scan acquisition in a Gabor-domain optical coherence microscopy (GD-OCM) instrument with a novel multilevel GPU architecture for high-speed imaging. GD-OCM yields high-definition volumetric imaging with dynamic depth of focusing through a bio-inspired liquid lens-based microscope design, which has no moving parts and is suitable for use in a manufacturing setting or in a medical environment. A dual-axis MEMS mirror was chosen to replace two single-axis galvanometer mirrors; as a result, the astigmatism caused by the mismatch between the optical pupil and the scanning location was eliminated and a 12x reduction in volume of the scanning system was achieved. Imaging at an invariant resolution of 2 μm was demonstrated throughout a volume of 1 × 1 × 0.6 mm3, acquired in less than 2 minutes. The MEMS-based scanner resulted in improved image quality, increased robustness and lighter weight of the system - all factors that are critical for on-field deployment. A custom integrated feedback system consisting of a laser diode and a position-sensing detector was developed to investigate the impact of the resonant frequency of the MEMS and the driving signal of the scanner on the movement of the mirror. Results on the metrology of manufactured materials and characterization of tissue samples with GD-OCM are presented.

  3. A compact acousto-optic lens for 2D and 3D femtosecond based 2-photon microscopy

    PubMed Central

    Kirkby, Paul A.; Naga Srinivas, N.K.M.; Silver, R. Angus

    2010-01-01

    We describe a high speed 3D Acousto-Optic Lens Microscope (AOLM) for femtosecond 2-photon imaging. By optimizing the design of the 4 AO Deflectors (AODs) and by deriving new control algorithms, we have developed a compact spherical AOL with a low temporal dispersion that enables 2-photon imaging at 10-fold lower power than previously reported. We show that the AOLM can perform high speed 2D raster-scan imaging (>150 Hz) without scan rate dependent astigmatism. It can deflect and focus a laser beam in a 3D random access sequence at 30 kHz and has an extended focusing range (>137 μm; 40X 0.8NA objective). These features are likely to make the AOLM a useful tool for studying fast physiological processes distributed in 3D space PMID:20588506

  4. 3D inclusion trail geometry determination within individual porphyroblasts using reflected light optical microscopy of oriented blocks

    NASA Astrophysics Data System (ADS)

    Munro, Mark; Bowden, Douglas; Ord, Alison; Hobbs, Bruce

    2015-04-01

    It is vital to interpret porphyroblast microstructures accurately relative to both one another and to external matrix structures when using them to reconstruct the tectono-metamorphic evolution of orogenic terranes. Mis-interpretation may have profound implications for either the deformation component or the inferred metamorphic reactions resulting in erroneous Pressure-Temperature-time-Deformation (P-T-t-D) trajectories. A number of well-established approaches have been devised for measuring porphyroblast inclusion trails including pitch and strike measurement, 'FitPitch' best-fit plane assignment, and the radial asymmetry method. A long-standing limitation of these methods is that they generally permit only a single measurement to be extracted from each individual porphyroblast, and therefore provide mean 3D orientation data for an entire population. Alternatively, High-Resolution X-ray Computed Tomography (HRXCT) facilitates the imaging of 3D internal geometries within individuals. However, at present significant operating costs render it unviable for routine application to large numbers of samples required for extracting meaningful tectonic interpretations. Here, a new method is presented for the determination of 3D geometries within porphyroblasts using reflected light examination of polished schist material. Reflected light microscopy yields good quality representation of inclusion trails preserved within porphyroblasts. Sectioning oriented samples into small, oriented blocks allows multiple intersections through porphyroblasts (generally >5mm) to be measured via mechanical stage and amalgamated to reconstruct the plane in 3D. The method represents an accessible alternative to HRXCT, which is applicable to any porphyroblastic phase of adequate size to permit at least two intersections. The technique is demonstrated on garnets from the Mesoproterozoic Mount Barren Group, southern Albany-Fraser orogen of S. W. Australia. Porphyroblasts within a structural

  5. 3DSEM: A 3D microscopy dataset.

    PubMed

    Tafti, Ahmad P; Kirkpatrick, Andrew B; Holz, Jessica D; Owen, Heather A; Yu, Zeyun

    2016-03-01

    The Scanning Electron Microscope (SEM) as a 2D imaging instrument has been widely used in many scientific disciplines including biological, mechanical, and materials sciences to determine the surface attributes of microscopic objects. However the SEM micrographs still remain 2D images. To effectively measure and visualize the surface properties, we need to truly restore the 3D shape model from 2D SEM images. Having 3D surfaces would provide anatomic shape of micro-samples which allows for quantitative measurements and informative visualization of the specimens being investigated. The 3DSEM is a dataset for 3D microscopy vision which is freely available at [1] for any academic, educational, and research purposes. The dataset includes both 2D images and 3D reconstructed surfaces of several real microscopic samples. PMID:26779561

  6. 3DSEM: A 3D microscopy dataset

    PubMed Central

    Tafti, Ahmad P.; Kirkpatrick, Andrew B.; Holz, Jessica D.; Owen, Heather A.; Yu, Zeyun

    2015-01-01

    The Scanning Electron Microscope (SEM) as a 2D imaging instrument has been widely used in many scientific disciplines including biological, mechanical, and materials sciences to determine the surface attributes of microscopic objects. However the SEM micrographs still remain 2D images. To effectively measure and visualize the surface properties, we need to truly restore the 3D shape model from 2D SEM images. Having 3D surfaces would provide anatomic shape of micro-samples which allows for quantitative measurements and informative visualization of the specimens being investigated. The 3DSEM is a dataset for 3D microscopy vision which is freely available at [1] for any academic, educational, and research purposes. The dataset includes both 2D images and 3D reconstructed surfaces of several real microscopic samples. PMID:26779561

  7. 3DSEM: A 3D microscopy dataset.

    PubMed

    Tafti, Ahmad P; Kirkpatrick, Andrew B; Holz, Jessica D; Owen, Heather A; Yu, Zeyun

    2016-03-01

    The Scanning Electron Microscope (SEM) as a 2D imaging instrument has been widely used in many scientific disciplines including biological, mechanical, and materials sciences to determine the surface attributes of microscopic objects. However the SEM micrographs still remain 2D images. To effectively measure and visualize the surface properties, we need to truly restore the 3D shape model from 2D SEM images. Having 3D surfaces would provide anatomic shape of micro-samples which allows for quantitative measurements and informative visualization of the specimens being investigated. The 3DSEM is a dataset for 3D microscopy vision which is freely available at [1] for any academic, educational, and research purposes. The dataset includes both 2D images and 3D reconstructed surfaces of several real microscopic samples.

  8. 3D differential phase contrast microscopy

    NASA Astrophysics Data System (ADS)

    Chen, Michael; Tian, Lei; Waller, Laura

    2016-03-01

    We demonstrate three-dimensional (3D) optical phase and amplitude reconstruction based on coded source illumination using a programmable LED array. Multiple stacks of images along the optical axis are computed from recorded intensities captured by multiple images under off-axis illumination. Based on the first Born approximation, a linear differential phase contrast (DPC) model is built between 3D complex index of refraction and the intensity stacks. Therefore, 3D volume reconstruction can be achieved via a fast inversion method, without the intermediate 2D phase retrieval step. Our system employs spatially partially coherent illumination, so the transverse resolution achieves twice the NA of coherent systems, while axial resolution is also improved 2× as compared to holographic imaging.

  9. 3D microscopy - new powerful tools in geomaterials characterization

    NASA Astrophysics Data System (ADS)

    Mauko Pranjić, Alenka; Mladenovič, Ana; Turk, Janez; Šajna, Aljoša; Čretnik, Janko

    2016-04-01

    Microtomography (microCT) is becoming more and more widely recognized in geological sciences as a powerful tool for the spatial characterization of rock and other geological materials. Together with 3D image analysis and other complementary techniques, it has the characteristics of an innovative and non-destructive 3D microscopical technique. On the other hand its main disadvantages are low availability (only a few geological laboratories are equipped with high resolution tomographs), the relatively high prices of testing connected with the use of an xray source, technical limitations connected to the resolution and imaging of certain materials, as well as timeconsuming and complex 3D image analysis, necessary for quantification of 3D tomographic data sets. In this work three examples are presented of optimal 3D microscopy analysis of geomaterials in construction such as porosity characterization of impregnated sandstone, aerated concrete and marble prone to bowing. Studies include processes of microCT imaging, 3D data analysis and fitting of data with complementary analysis, such as confocal microscopy, mercury porosimetry, gas sorption, optical/fluorescent microscopy and scanning electron microscopy. Present work has been done in the frame of national research project 3D and 4D microscopy development of new powerful tools in geosciences (ARRS J1-7148) funded by Slovenian Research Agency.

  10. Axial Plane Optical Microscopy

    PubMed Central

    Li, Tongcang; Ota, Sadao; Kim, Jeongmin; Wong, Zi Jing; Wang, Yuan; Yin, Xiaobo; Zhang, Xiang

    2014-01-01

    We present axial plane optical microscopy (APOM) that can, in contrast to conventional microscopy, directly image a sample's cross-section parallel to the optical axis of an objective lens without scanning. APOM combined with conventional microscopy simultaneously provides two orthogonal images of a 3D sample. More importantly, APOM uses only a single lens near the sample to achieve selective-plane illumination microscopy, as we demonstrated by three-dimensional (3D) imaging of fluorescent pollens and brain slices. This technique allows fast, high-contrast, and convenient 3D imaging of structures that are hundreds of microns beneath the surfaces of large biological tissues. PMID:25434770

  11. Axial Plane Optical Microscopy

    NASA Astrophysics Data System (ADS)

    Li, Tongcang; Ota, Sadao; Kim, Jeongmin; Wong, Zi Jing; Wang, Yuan; Yin, Xiaobo; Zhang, Xiang

    2014-12-01

    We present axial plane optical microscopy (APOM) that can, in contrast to conventional microscopy, directly image a sample's cross-section parallel to the optical axis of an objective lens without scanning. APOM combined with conventional microscopy simultaneously provides two orthogonal images of a 3D sample. More importantly, APOM uses only a single lens near the sample to achieve selective-plane illumination microscopy, as we demonstrated by three-dimensional (3D) imaging of fluorescent pollens and brain slices. This technique allows fast, high-contrast, and convenient 3D imaging of structures that are hundreds of microns beneath the surfaces of large biological tissues.

  12. 3D microscopy for microfabrication quality control

    NASA Astrophysics Data System (ADS)

    Muller, Matthew S.; De Jean, Paul D.

    2015-03-01

    A novel stereo microscope adapter, the SweptVue, has been developed to rapidly perform quantitative 3D microscopy for cost-effective microfabrication quality control. The SweptVue adapter uses the left and right stereo channels of an Olympus SZX7 stereo microscope for sample illumination and detection, respectively. By adjusting the temporal synchronization between the illumination lines projected from a Texas Instruments DLP LightCrafter and the rolling shutter on a Point Grey Flea3 CMOS camera, micrometer-scale depth features can be easily and rapidly measured at up to 5 μm resolution on a variety of microfabricated samples. In this study, the build performance of an industrial-grade Stratasys Object 300 Connex 3D printer was examined. Ten identical parts were 3D printed with a lateral and depth resolution of 42 μm and 30 μm, respectively, using both a rigid and flexible Stratasys PolyJet material. Surface elevation precision and accuracy was examined over multiple regions of interest on plateau and hemispherical surfaces. In general, the dimensions of the examined features were reproducible across the parts built using both materials. However, significant systemic lateral and height build errors were discovered, such as: decreased heights when approaching the edges of plateaus, inaccurate height steps, and poor tolerances on channel width. For 3D printed parts to be used in functional applications requiring micro-scale tolerances, they need to conform to specification. Despite appearing identical, our 3D printed parts were found to have a variety of defects that the SweptVue adapter quickly revealed.

  13. Validation of image processing tools for 3-D fluorescence microscopy.

    PubMed

    Dieterlen, Alain; Xu, Chengqi; Gramain, Marie-Pierre; Haeberlé, Olivier; Colicchio, Bruno; Cudel, Christophe; Jacquey, Serge; Ginglinger, Emanuelle; Jung, Georges; Jeandidier, Eric

    2002-04-01

    3-D optical fluorescent microscopy becomes nowadays an efficient tool for volumic investigation of living biological samples. Using optical sectioning technique, a stack of 2-D images is obtained. However, due to the nature of the system optical transfer function and non-optimal experimental conditions, acquired raw data usually suffer from some distortions. In order to carry out biological analysis, raw data have to be restored by deconvolution. The system identification by the point-spread function is useful to obtain the knowledge of the actual system and experimental parameters, which is necessary to restore raw data. It is furthermore helpful to precise the experimental protocol. In order to facilitate the use of image processing techniques, a multi-platform-compatible software package called VIEW3D has been developed. It integrates a set of tools for the analysis of fluorescence images from 3-D wide-field or confocal microscopy. A number of regularisation parameters for data restoration are determined automatically. Common geometrical measurements and morphological descriptors of fluorescent sites are also implemented to facilitate the characterisation of biological samples. An example of this method concerning cytogenetics is presented.

  14. 3D electron microscopy of biological nanomachines: principles and applications.

    PubMed

    Sorzano, C O S; Jonic, S; Cottevieille, M; Larquet, E; Boisset, N; Marco, S

    2007-11-01

    Transmission electron microscopy is a powerful technique for studying the three-dimensional (3D) structure of a wide range of biological specimens. Knowledge of this structure is crucial for fully understanding complex relationships among macromolecular complexes and organelles in living cells. In this paper, we present the principles and main application domains of 3D transmission electron microscopy in structural biology. Moreover, we survey current developments needed in this field, and discuss the close relationship of 3D transmission electron microscopy with other experimental techniques aimed at obtaining structural and dynamical information from the scale of whole living cells to atomic structure of macromolecular complexes.

  15. Sample drift correction in 3D fluorescence photoactivation localization microscopy

    NASA Astrophysics Data System (ADS)

    Mlodzianoski, Michael J.; Schreiner, John M.; Callahan, Steven P.; Smolková, Katarina; Dlasková, Andrea; Šantorová, Jitka; Ježek, Petr; Bewersdorf, Joerg

    2011-08-01

    The recent development of diffraction-unlimited far-field fluorescence microscopy has overcome the classical resolution limit of ~250 nm of conventional light microscopy by about a factor of ten. The improved resolution, however, reveals not only biological structures at an unprecedented resolution, but is also susceptible to sample drift on a much finer scale than previously relevant. Without correction, sample drift leads to smeared images with decreased resolution, and in the worst case to misinterpretation of the imaged structures. This poses a problem especially for techniques such as Fluorescence Photoactivation Localization Microscopy (FPALM/PALM) or Stochastic Optical Reconstruction Microscopy (STORM), which often require minutes recording time. Here we discuss an approach that corrects for three-dimensional (3D) drift in images of fixed samples without the requirement for fiduciary markers or instrument modifications. Drift is determined by calculating the spatial cross-correlation function between subsets of localized particles imaged at different times. Correction down to ~5 nm precision is achieved despite the fact that different molecules are imaged in each frame. We demonstrate the performance of our drift correction algorithm with different simulated structures and analyze its dependence on particle density and localization precision. By imaging mitochondria with Biplane FPALM we show our algorithm's feasibility in a practical application.

  16. FIT3D: Fitting optical spectra

    NASA Astrophysics Data System (ADS)

    Sánchez, S. F.; Pérez, E.; Sánchez-Blázquez, P.; González, J. J.; Rosales-Ortega, F. F.; Cano-Díaz, M.; López-Cobá, C.; Marino, R. A.; Gil de Paz, A.; Mollá, M.; López-Sánchez, A. R.; Ascasibar, Y.; Barrera-Ballesteros, J.

    2016-09-01

    FIT3D fits optical spectra to deblend the underlying stellar population and the ionized gas, and extract physical information from each component. FIT3D is focused on the analysis of Integral Field Spectroscopy data, but is not restricted to it, and is the basis of Pipe3D, a pipeline used in the analysis of datasets like CALIFA, MaNGA, and SAMI. It can run iteratively or in an automatic way to derive the parameters of a large set of spectra.

  17. Towards Single Cell Traction Microscopy within 3D Collagen Matrices

    PubMed Central

    Hall, Matthew S.; Long, Rong; Feng, Xinzeng; Huang, YuLing; Hui, Chung-Yuen; Wu, Mingming

    2013-01-01

    Mechanical interaction between the cell and its extracellular matrix (ECM) regulates cellular behaviors, including proliferation, differentiation, adhesion, and migration. Cells require the three dimensional (3D) architectural support of the ECM to perform physiologically realistic functions. However, current understanding of cell-ECM and cell-cell mechanical interactions is largely derived from 2D cell traction force microscopy, in which cells are cultured on a flat substrate. 3D cell traction microscopy is emerging for mapping traction fields of single animal cells embedded in either synthetic or natively derived fibrous gels. We discuss here the development of 3D cell traction microscopy, its current limitations, and perspectives on the future of this technology. Emphasis is placed on strategies for applying 3D cell traction microscopy to individual tumor cells migration within collagen gels. PMID:23806281

  18. Microscopy in 3D: a biologist’s toolbox

    PubMed Central

    Fischer, Robert S.; Wu, Yicong; Kanchanawong, Pakorn; Shroff, Hari; Waterman, Clare M.

    2012-01-01

    The power of fluorescence microscopy to study cellular structures and macromolecular complexes spans a wide range of size scales, from studies of cell behavior and function in physiological, three-dimensional (3D) environments, to understanding the molecular architecture of organelles. At each length scale, the challenge in 3D imaging is to extract the most spatial and temporal resolution possible while limiting photodamage/bleaching to living cells. A number of advancements in 3D fluorescence microscopy now offer higher resolution, improved speed, and reduced photobleaching relative to traditional point-scanning microscopy methods. Here, we discuss a few specific microscopy modalities that we believe will be particularly advantageous in imaging cells and subcellular structures in physiologically relevant 3D environments. PMID:22047760

  19. 3D imaging of neutron tracks using confocal microscopy

    NASA Astrophysics Data System (ADS)

    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

  20. Toward single cell traction microscopy within 3D collagen matrices

    SciTech Connect

    Hall, Matthew S.; Long, Rong; Feng, Xinzeng; Huang, YuLing; Hui, Chung-Yuen; Wu, Mingming

    2013-10-01

    Mechanical interaction between the cell and its extracellular matrix (ECM) regulates cellular behaviors, including proliferation, differentiation, adhesion, and migration. Cells require the three-dimensional (3D) architectural support of the ECM to perform physiologically realistic functions. However, current understanding of cell–ECM and cell–cell mechanical interactions is largely derived from 2D cell traction force microscopy, in which cells are cultured on a flat substrate. 3D cell traction microscopy is emerging for mapping traction fields of single animal cells embedded in either synthetic or natively derived fibrous gels. We discuss here the development of 3D cell traction microscopy, its current limitations, and perspectives on the future of this technology. Emphasis is placed on strategies for applying 3D cell traction microscopy to individual tumor cell migration within collagen gels. - Highlights: • Review of the current state of the art in 3D cell traction force microscopy. • Bulk and micro-characterization of remodelable fibrous collagen gels. • Strategies for performing 3D cell traction microscopy within collagen gels.

  1. Fringe projection 3D microscopy with the general imaging model.

    PubMed

    Yin, Yongkai; Wang, Meng; Gao, Bruce Z; Liu, Xiaoli; Peng, Xiang

    2015-03-01

    Three-dimensional (3D) imaging and metrology of microstructures is a critical task for the design, fabrication, and inspection of microelements. Newly developed fringe projection 3D microscopy is presented in this paper. The system is configured according to camera-projector layout and long working distance lenses. The Scheimpflug principle is employed to make full use of the limited depth of field. For such a specific system, the general imaging model is introduced to reach a full 3D reconstruction. A dedicated calibration procedure is developed to realize quantitative 3D imaging. Experiments with a prototype demonstrate the accessibility of the proposed configuration, model, and calibration approach.

  2. 3D optical measuring technologies and systems

    NASA Astrophysics Data System (ADS)

    Chugui, Yuri V.

    2005-02-01

    The results of the R & D activity of TDI SIE SB RAS in the field of the 3D optical measuring technologies and systems for noncontact 3D optical dimensional inspection applied to atomic and railway industry safety problems are presented. This activity includes investigations of diffraction phenomena on some 3D objects, using the original constructive calculation method. The efficient algorithms for precise determining the transverse and longitudinal sizes of 3D objects of constant thickness by diffraction method, peculiarities on formation of the shadow and images of the typical elements of the extended objects were suggested. Ensuring the safety of nuclear reactors and running trains as well as their high exploitation reliability requires a 100% noncontact precise inspection of geometrical parameters of their components. To solve this problem we have developed methods and produced the technical vision measuring systems LMM, CONTROL, PROFIL, and technologies for noncontact 3D dimensional inspection of grid spacers and fuel elements for the nuclear reactor VVER-1000 and VVER-440, as well as automatic laser diagnostic COMPLEX for noncontact inspection of geometric parameters of running freight car wheel pairs. The performances of these systems and the results of industrial testing are presented and discussed. The created devices are in pilot operation at Atomic and Railway Companies.

  3. Astigmatic multifocus microscopy enables deep 3D super-resolved imaging

    PubMed Central

    Oudjedi, Laura; Fiche, Jean-Bernard; Abrahamsson, Sara; Mazenq, Laurent; Lecestre, Aurélie; Calmon, Pierre-François; Cerf, Aline; Nöllmann, Marcelo

    2016-01-01

    We have developed a 3D super-resolution microscopy method that enables deep imaging in cells. This technique relies on the effective combination of multifocus microscopy and astigmatic 3D single-molecule localization microscopy. We describe the optical system and the fabrication process of its key element, the multifocus grating. Then, two strategies for localizing emitters with our imaging method are presented and compared with a previously described deep 3D localization algorithm. Finally, we demonstrate the performance of the method by imaging the nuclear envelope of eukaryotic cells reaching a depth of field of ~4µm. PMID:27375935

  4. Astigmatic multifocus microscopy enables deep 3D super-resolved imaging.

    PubMed

    Oudjedi, Laura; Fiche, Jean-Bernard; Abrahamsson, Sara; Mazenq, Laurent; Lecestre, Aurélie; Calmon, Pierre-François; Cerf, Aline; Nöllmann, Marcelo

    2016-06-01

    We have developed a 3D super-resolution microscopy method that enables deep imaging in cells. This technique relies on the effective combination of multifocus microscopy and astigmatic 3D single-molecule localization microscopy. We describe the optical system and the fabrication process of its key element, the multifocus grating. Then, two strategies for localizing emitters with our imaging method are presented and compared with a previously described deep 3D localization algorithm. Finally, we demonstrate the performance of the method by imaging the nuclear envelope of eukaryotic cells reaching a depth of field of ~4µm.

  5. Holographic microscopy for 3D tracking of bacteria

    NASA Astrophysics Data System (ADS)

    Nadeau, Jay; Cho, Yong Bin; El-Kholy, Marwan; Bedrossian, Manuel; Rider, Stephanie; Lindensmith, Christian; Wallace, J. Kent

    2016-03-01

    Understanding when, how, and if bacteria swim is key to understanding critical ecological and biological processes, from carbon cycling to infection. Imaging motility by traditional light microscopy is limited by focus depth, requiring cells to be constrained in z. Holographic microscopy offers an instantaneous 3D snapshot of a large sample volume, and is therefore ideal in principle for quantifying unconstrained bacterial motility. However, resolving and tracking individual cells is difficult due to the low amplitude and phase contrast of the cells; the index of refraction of typical bacteria differs from that of water only at the second decimal place. In this work we present a combination of optical and sample-handling approaches to facilitating bacterial tracking by holographic phase imaging. The first is the design of the microscope, which is an off-axis design with the optics along a common path, which minimizes alignment issues while providing all of the advantages of off-axis holography. Second, we use anti-reflective coated etalon glass in the design of sample chambers, which reduce internal reflections. Improvement seen with the antireflective coating is seen primarily in phase imaging, and its quantification is presented here. Finally, dyes may be used to increase phase contrast according to the Kramers-Kronig relations. Results using three test strains are presented, illustrating the different types of bacterial motility characterized by an enteric organism (Escherichia coli), an environmental organism (Bacillus subtilis), and a marine organism (Vibrio alginolyticus). Data processing steps to increase the quality of the phase images and facilitate tracking are also discussed.

  6. 3D super-resolution microscopy of bacterial division machinery

    NASA Astrophysics Data System (ADS)

    Vedyaykin, A. D.; Sabantsev, A. V.; Vishnyakov, I. E.; Morozova, N. E.; Polinovskaya, V. S.; Khodorkovskii, M. A.

    2016-08-01

    Super-resolution microscopy is a promising tool for the field of microbiology, as bacteria sizes are comparable to the resolution limit of light microscopy. Bacterial division machinery and FtsZ protein in particular attract much attention of scientists who use different super-resolution microscopy techniques, but most of the available data on FtsZ structures was obtained using two-dimensional (2D) super-resolution microscopy. Using 3D single-molecule localization microscopy (SMLM, namely dSTORM) to visualize FtsZ, we demonstrate that this approach allows more accurate interpretation of super-resolution images and provides new opportunities for the study of complex structures like bacterial divisome.

  7. Applied 3D printing for microscopy in health science research

    NASA Astrophysics Data System (ADS)

    Brideau, Craig; Zareinia, Kourosh; Stys, Peter

    2015-03-01

    The rapid prototyping capability offered by 3D printing is considered advantageous for commercial applications. However, the ability to quickly produce precision custom devices is highly beneficial in the research laboratory setting as well. Biological laboratories require the manipulation and analysis of delicate living samples, thus the ability to create custom holders, support equipment, and adapters allow the extension of existing laboratory machines. Applications include camera adapters and stage sample holders for microscopes, surgical guides for tissue preparation, and small precision tools customized to unique specifications. Where high precision is needed, especially the reproduction of fine features, a printer with a high resolution is needed. However, the introduction of cheaper, lower resolution commercial printers have been shown to be more than adequate for less demanding projects. For direct manipulation of delicate samples, biocompatible raw materials are often required, complicating the printing process. This paper will examine some examples of 3D-printed objects for laboratory use, and provide an overview of the requirements for 3D printing for this application. Materials, printing resolution, production, and ease of use will all be reviewed with an eye to producing better printers and techniques for laboratory applications. Specific case studies will highlight applications for 3D-printed devices in live animal imaging for both microscopy and Magnetic Resonance Imaging.

  8. Shaping Field for 3D Laser Scanning Microscopy

    PubMed Central

    Colon, Jorge; Lim, Hyungsik

    2015-01-01

    Imaging deep tissue can be extremely inefficient when the region of interest is non-planar and buried in a thick sample, yielding a severely limited effective field of view (FOV). Here we describe a novel technique, namely adaptive field microscopy, which improves the efficiency of 3D imaging by controlling the image plane. The plane of scanning laser focus is continuously reshaped in situ to match the conformation of the sample. The practicality is demonstrated for ophthalmic imaging, where a large area of the corneal epithelium of intact mouse eye is captured in a single frame with subcellular resolution. PMID:26176454

  9. Diffractive optical element for creating visual 3D images.

    PubMed

    Goncharsky, Alexander; Goncharsky, Anton; Durlevich, Svyatoslav

    2016-05-01

    A method is proposed to compute and synthesize the microrelief of a diffractive optical element to produce a new visual security feature - the vertical 3D/3D switch effect. The security feature consists in the alternation of two 3D color images when the diffractive element is tilted up/down. Optical security elements that produce the new security feature are synthesized using electron-beam technology. Sample optical security elements are manufactured that produce 3D to 3D visual switch effect when illuminated by white light. Photos and video records of the vertical 3D/3D switch effect of real optical elements are presented. The optical elements developed can be replicated using standard equipment employed for manufacturing security holograms. The new optical security feature is easy to control visually, safely protected against counterfeit, and designed to protect banknotes, documents, ID cards, etc. PMID:27137530

  10. Calibration of optical 3D-measuring instruments

    NASA Astrophysics Data System (ADS)

    Brand, Uwe

    1998-09-01

    For the inspection and measurement of microstructures small accurate three-dimensional coordinate measuring machines are needed. Typical measurement volumes are 10 mm by 10 mm by 3 mm and the desired 3D-measurement uncertainty is 0.1 micrometer. Up to now only optical coordinate measuring machines (CMM) offer the necessary lateral measurement ranges. But optical CMMs are restricted to two-dimensional measurements and moreover the aimed uncertainty has not been achieved yet. Since a few years new optical techniques are available which are able to measure nearly three-dimensionally (scanning white light, fringe projection, confocal microscopy, photogrammetry). In order to use these instruments and to specify their measurement uncertainty, calibration of these instruments is necessary. The calibration of the three measurement axes is divided into calibration of the lateral axes and calibration of the vertical axis. The contribution focuses on the development of new depth setting standards (1 micrometer - 1 milimeter) and their traceability.

  11. Resolution improvement by 3D particle averaging in localization microscopy

    PubMed Central

    Broeken, Jordi; Johnson, Hannah; Lidke, Diane S.; Liu, Sheng; Nieuwenhuizen, Robert P.J.; Stallinga, Sjoerd; Lidke, Keith A.; Rieger, Bernd

    2015-01-01

    Inspired by recent developments in localization microscopy that applied averaging of identical particles in 2D for increasing the resolution even further, we discuss considerations for alignment (registration) methods for particles in general and for 3D in particular. We detail that traditional techniques for particle registration from cryo electron microscopy based on cross-correlation are not suitable, as the underlying image formation process is fundamentally different. We argue that only localizations, i.e. a set of coordinates with associated uncertainties, are recorded and not a continuous intensity distribution. We present a method that owes to this fact and that is inspired by the field of statistical pattern recognition. In particular we suggest to use an adapted version of the Bhattacharyya distance as a merit function for registration. We evaluate the method in simulations and demonstrate it on three-dimensional super-resolution data of Alexa 647 labelled to the Nup133 protein in the nuclear pore complex of Hela cells. From the simulations we find suggestions that for successful registration the localization uncertainty must be smaller than the distance between labeling sites on a particle. These suggestions are supported by theoretical considerations concerning the attainable resolution in localization microscopy and its scaling behavior as a function of labeling density and localization precision. PMID:25866640

  12. Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy

    PubMed Central

    Changou, Chun A.; Wolfson, Deanna L.; Ahluwalia, Balpreet Singh; Bold, Richard J.; Kung, Hsing-Jien; Chuang, Frank Y.S.

    2013-01-01

    Prostate cancer is the leading form of malignancies among men in the U.S. While surgery carries a significant risk of impotence and incontinence, traditional chemotherapeutic approaches have been largely unsuccessful. Hormone therapy is effective at early stage, but often fails with the eventual development of hormone-refractory tumors. We have been interested in developing therapeutics targeting specific metabolic deficiency of tumor cells. We recently showed that prostate tumor cells specifically lack an enzyme (argininosuccinate synthase, or ASS) involved in the synthesis of the amino acid arginine1. This condition causes the tumor cells to become dependent on exogenous arginine, and they undergo metabolic stress when free arginine is depleted by arginine deiminase (ADI)1,10. Indeed, we have shown that human prostate cancer cells CWR22Rv1 are effectively killed by ADI with caspase-independent apoptosis and aggressive autophagy (or macroautophagy)1,2,3. Autophagy is an evolutionarily-conserved process that allows cells to metabolize unwanted proteins by lysosomal breakdown during nutritional starvation4,5. Although the essential components of this pathway are well-characterized6,7,8,9, many aspects of the molecular mechanism are still unclear - in particular, what is the role of autophagy in the death-response of prostate cancer cells after ADI treatment? In order to address this question, we required an experimental method to measure the level and extent of autophagic response in cells - and since there are no known molecular markers that can accurately track this process, we chose to develop an imaging-based approach, using quantitative 3D fluorescence microscopy11,12. Using CWR22Rv1 cells specifically-labeled with fluorescent probes for autophagosomes and lysosomes, we show that 3D image stacks acquired with either widefield deconvolution microscopy (and later, with super-resolution, structured-illumination microscopy) can clearly capture the early stages of

  13. Quantitative analysis of autophagy using advanced 3D fluorescence microscopy.

    PubMed

    Changou, Chun A; Wolfson, Deanna L; Ahluwalia, Balpreet Singh; Bold, Richard J; Kung, Hsing-Jien; Chuang, Frank Y S

    2013-01-01

    Prostate cancer is the leading form of malignancies among men in the U.S. While surgery carries a significant risk of impotence and incontinence, traditional chemotherapeutic approaches have been largely unsuccessful. Hormone therapy is effective at early stage, but often fails with the eventual development of hormone-refractory tumors. We have been interested in developing therapeutics targeting specific metabolic deficiency of tumor cells. We recently showed that prostate tumor cells specifically lack an enzyme (argininosuccinate synthase, or ASS) involved in the synthesis of the amino acid arginine(1). This condition causes the tumor cells to become dependent on exogenous arginine, and they undergo metabolic stress when free arginine is depleted by arginine deiminase (ADI)(1,10). Indeed, we have shown that human prostate cancer cells CWR22Rv1 are effectively killed by ADI with caspase-independent apoptosis and aggressive autophagy (or macroautophagy)(1,2,3). Autophagy is an evolutionarily-conserved process that allows cells to metabolize unwanted proteins by lysosomal breakdown during nutritional starvation(4,5). Although the essential components of this pathway are well-characterized(6,7,8,9), many aspects of the molecular mechanism are still unclear - in particular, what is the role of autophagy in the death-response of prostate cancer cells after ADI treatment? In order to address this question, we required an experimental method to measure the level and extent of autophagic response in cells - and since there are no known molecular markers that can accurately track this process, we chose to develop an imaging-based approach, using quantitative 3D fluorescence microscopy(11,12). Using CWR22Rv1 cells specifically-labeled with fluorescent probes for autophagosomes and lysosomes, we show that 3D image stacks acquired with either widefield deconvolution microscopy (and later, with super-resolution, structured-illumination microscopy) can clearly capture the early

  14. Quantitative analysis of autophagy using advanced 3D fluorescence microscopy.

    PubMed

    Changou, Chun A; Wolfson, Deanna L; Ahluwalia, Balpreet Singh; Bold, Richard J; Kung, Hsing-Jien; Chuang, Frank Y S

    2013-05-03

    Prostate cancer is the leading form of malignancies among men in the U.S. While surgery carries a significant risk of impotence and incontinence, traditional chemotherapeutic approaches have been largely unsuccessful. Hormone therapy is effective at early stage, but often fails with the eventual development of hormone-refractory tumors. We have been interested in developing therapeutics targeting specific metabolic deficiency of tumor cells. We recently showed that prostate tumor cells specifically lack an enzyme (argininosuccinate synthase, or ASS) involved in the synthesis of the amino acid arginine(1). This condition causes the tumor cells to become dependent on exogenous arginine, and they undergo metabolic stress when free arginine is depleted by arginine deiminase (ADI)(1,10). Indeed, we have shown that human prostate cancer cells CWR22Rv1 are effectively killed by ADI with caspase-independent apoptosis and aggressive autophagy (or macroautophagy)(1,2,3). Autophagy is an evolutionarily-conserved process that allows cells to metabolize unwanted proteins by lysosomal breakdown during nutritional starvation(4,5). Although the essential components of this pathway are well-characterized(6,7,8,9), many aspects of the molecular mechanism are still unclear - in particular, what is the role of autophagy in the death-response of prostate cancer cells after ADI treatment? In order to address this question, we required an experimental method to measure the level and extent of autophagic response in cells - and since there are no known molecular markers that can accurately track this process, we chose to develop an imaging-based approach, using quantitative 3D fluorescence microscopy(11,12). Using CWR22Rv1 cells specifically-labeled with fluorescent probes for autophagosomes and lysosomes, we show that 3D image stacks acquired with either widefield deconvolution microscopy (and later, with super-resolution, structured-illumination microscopy) can clearly capture the early

  15. Spatially varying regularization of deconvolution in 3D microscopy.

    PubMed

    Seo, J; Hwang, S; Lee, J-M; Park, H

    2014-08-01

    Confocal microscopy has become an essential tool to explore biospecimens in 3D. Confocal microcopy images are still degraded by out-of-focus blur and Poisson noise. Many deconvolution methods including the Richardson-Lucy (RL) method, Tikhonov method and split-gradient (SG) method have been well received. The RL deconvolution method results in enhanced image quality, especially for Poisson noise. Tikhonov deconvolution method improves the RL method by imposing a prior model of spatial regularization, which encourages adjacent voxels to appear similar. The SG method also contains spatial regularization and is capable of incorporating many edge-preserving priors resulting in improved image quality. The strength of spatial regularization is fixed regardless of spatial location for the Tikhonov and SG method. The Tikhonov and the SG deconvolution methods are improved upon in this study by allowing the strength of spatial regularization to differ for different spatial locations in a given image. The novel method shows improved image quality. The method was tested on phantom data for which ground truth and the point spread function are known. A Kullback-Leibler (KL) divergence value of 0.097 is obtained with applying spatially variable regularization to the SG method, whereas KL value of 0.409 is obtained with the Tikhonov method. In tests on a real data, for which the ground truth is unknown, the reconstructed data show improved noise characteristics while maintaining the important image features such as edges.

  16. Manufacturing: 3D printed micro-optics

    NASA Astrophysics Data System (ADS)

    Juodkazis, Saulius

    2016-08-01

    Uncompromised performance of micro-optical compound lenses has been achieved by high-fidelity shape definition during two-photon absorption microfabrication. The lenses have been made directly onto image sensors and even onto the tip of an optic fibre.

  17. Infrared differential interference contrast microscopy for 3D interconnect overlay metrology.

    PubMed

    Ku, Yi-sha; Shyu, Deh-Ming; Lin, Yeou-Sung; Cho, Chia-Hung

    2013-08-12

    One of the main challenges for 3D interconnect metrology of bonded wafers is measuring through opaque silicon wafers using conventional optical microscopy. We demonstrate here the use infrared microscopy, enhanced by implementing the differential interference contrast (DIC) technique, to measure the wafer bonding overlay. A pair of two dimensional symmetric overlay marks were processed at both the front and back sides of thinned wafers to evaluate the bonding overlay. A self-developed analysis algorithm and theoretical fitting model was used to map the overlay error between the bonded wafers and the interconnect structures. The measurement accuracy was found to be better than 1.0 micron.

  18. Coherent Microscopy for 3-D Movement Monitoring and Super-Resolved Imaging

    NASA Astrophysics Data System (ADS)

    Beiderman, Yevgeny; Amsel, Avigail; Tzadka, Yaniv; Fixler, Dror; Teicher, Mina; Micó, Vicente; Garcí, Javier; Javidi, Bahram; DaneshPanah, Mehdi; Moon, Inkyu; Zalevsky, Zeev

    In this chapter we present three types of microscopy-related configurations while the first one is used for 3-D movement monitoring of the inspected samples, the second one is used for super-resolved 3-D imaging, and the last one presents an overview digital holographic microscopy applications. The first configuration is based on temporal tracking of secondary reflected speckles when imaged by properly defocused optics. We validate the proposed scheme by using it to monitor 3-D spontaneous contraction of rat's cardiac muscle cells while allowing nanometric tracking accuracy without interferometric recording. The second configuration includes projection of temporally varying speckle patterns on top of the sample and by proper decoding exceeding the diffraction as well as the geometrical-related lateral resolution limitation. In the final part of the chapter, we overview applications of digital holographic microscopy (DHM) for real-time non-invasive 3-D sensing, tracking, and recognition of living microorganisms such as single- or multiple-cell organisms and bacteria.

  19. 3D-additive manufactured optical mount

    NASA Astrophysics Data System (ADS)

    Mammini, Paul V.; Ciscel, David; Wooten, John

    2015-09-01

    The Area Defense Anti-Munitions (ADAM) is a low cost and effective high power laser weapon system. It's designed to address and negate important threats such as short-range rockets, UAVs, and small boats. Many critical optical components operate in the system. The optics and mounts must accommodate thermal and mechanical stresses, plus maintain an exceptional wave front during operation. Lockheed Martin Space Systems Company (LMSSC) developed, designed, and currently operates ADAM. This paper covers the design and development of a key monolithic, flexured, titanium mirror mount that was manufactured by CalRAM using additive processes.

  20. 3D printed long period gratings for optical fibers.

    PubMed

    Iezzi, Victor Lambin; Boisvert, Jean-Sébastien; Loranger, Sébastien; Kashyap, Raman

    2016-04-15

    We demonstrate a simple technique for implementing long period grating (LPG) structures by the use of a 3D printer. This Letter shows a way of manipulating the mode coupling within an optical fiber by applying stress through an external 3D printed periodic structure. Different LPG lengths and periods have been studied, as well as the effect of the applied stress on the coupling efficiency from the fundamental mode to cladding modes. The technique is very simple, highly flexible, affordable, and easy to implement without the need of altering the optical fiber. This Letter is part of a growing line of interest in the use of 3D printers for optical applications.

  1. 3-D stimulated emission depletion microscopy with programmable aberration correction.

    PubMed

    Lenz, Martin O; Sinclair, Hugo G; Savell, Alexander; Clegg, James H; Brown, Alice C N; Davis, Daniel M; Dunsby, Chris; Neil, Mark A A; French, Paul M W

    2014-01-01

    We present a stimulated emission depletion (STED) microscope that provides 3-D super resolution by simultaneous depletion using beams with both a helical phase profile for enhanced lateral resolution and an annular phase profile to enhance axial resolution. The 3-D depletion point spread function is realised using a single spatial light modulator that can also be programmed to compensate for aberrations in the microscope and the sample. We apply it to demonstrate the first 3-D super-resolved imaging of an immunological synapse between a Natural Killer cell and its target cell.

  2. Optical characterization and measurements of autostereoscopic 3D displays

    NASA Astrophysics Data System (ADS)

    Salmimaa, Marja; Järvenpää, Toni

    2008-04-01

    3D or autostereoscopic display technologies offer attractive solutions for enriching the multimedia experience. However, both characterization and comparison of 3D displays have been challenging when the definitions for the consistent measurement methods have been lacking and displays with similar specifications may appear quite different. Earlier we have investigated how the optical properties of autostereoscopic (3D) displays can be objectively measured and what are the main characteristics defining the perceived image quality. In this paper the discussion is extended to cover the viewing freedom (VF) and the definition for the optimum viewing distance (OVD) is elaborated. VF is the volume inside which the eyes have to be to see an acceptable 3D image. Characteristics limiting the VF space are proposed to be 3D crosstalk, luminance difference and color difference. Since the 3D crosstalk can be presumed to be dominating the quality of the end user experience and in our approach is forming the basis for the calculations of the other optical parameters, the reliability of the 3D crosstalk measurements is investigated. Furthermore the effect on the derived VF definition is evaluated. We have performed comparison 3D crosstalk measurements with different measurement device apertures and the effect of different measurement geometry on the results on actual 3D displays is reported.

  3. A new 3D tracking method exploiting the capabilities of digital holography in microscopy

    NASA Astrophysics Data System (ADS)

    Miccio, L.; Memmolo, P.; Merola, F.; Fusco, S.; Embrione, V.; Netti, P. A.; Ferraro, P.

    2013-04-01

    A method for 3D tracking has been developed exploiting Digital Holographic Microscopy (DHM) features. In the framework of self-consistent platform for manipulation and measurement of biological specimen we use DHM for quantitative and completely label free analysis of specimen with low amplitude contrast. Tracking capability extend the potentiality of DHM allowing to monitor the motion of appropriate probes and correlate it with sample properties. Complete 3D tracking has been obtained for the probes avoiding the issue of amplitude refocusing in traditional tracking processing. Our technique belongs to the video tracking methods that, conversely from Quadrant Photo-Diode method, opens the possibility to track multiples probes. All the common used video tracking algorithms are based on the numerical analysis of amplitude images in the focus plane and the shift of the maxima in the image plane are measured after the application of an appropriate threshold. Our approach for video tracking uses different theoretical basis. A set of interferograms is recorded and the complex wavefields are managed numerically to obtain three dimensional displacements of the probes. The procedure works properly on an higher number of probes and independently from their size. This method overcomes the traditional video tracking issues as the inability to measure the axial movement and the choice of suitable threshold mask. The novel configuration allows 3D tracking of micro-particles and simultaneously can furnish Quantitative Phase-contrast maps of tracked micro-objects by interference microscopy, without changing the configuration. In this paper, we show a new concept for a compact interferometric microscope that can ensure the multifunctionality, accomplishing accurate 3D tracking and quantitative phase-contrast analysis. Experimental results are presented and discussed for in vitro cells. Through a very simple and compact optical arrangement we show how two different functionalities

  4. 3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis.

    PubMed

    Park, Ok Kyu; Kwak, Jina; Jung, Yoo Jung; Kim, Young Ho; Hong, Hyun-Seok; Hwang, Byung Joon; Kwon, Seung-Hae; Kee, Yun

    2015-11-01

    Precise 3D spatial mapping of cells and their connections within living tissues is required to fully understand developmental processes and neural activities. Zebrafish embryos are relatively small and optically transparent, making them the vertebrate model of choice for live in vivo imaging. However, embryonic brains cannot be imaged in their entirety by confocal or two-photon microscopy due to limitations in optical range and scanning speed. Here, we use light-sheet fluorescence microscopy to overcome these limitations and image the entire head of live transgenic zebrafish embryos. We simultaneously imaged cranial neurons and blood vessels during embryogenesis, generating comprehensive 3D maps that provide insight into the coordinated morphogenesis of the nervous system and vasculature during early development. In addition, blood cells circulating through the entire head, vagal and cardiac vasculature were also visualized at high resolution in a 3D movie. These data provide the foundation for the construction of a complete 4D atlas of zebrafish embryogenesis and neural activity.

  5. 3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis

    PubMed Central

    Park, Ok Kyu; Kwak, Jina; Jung, Yoo Jung; Kim, Young Ho; Hong, Hyun-Seok; Hwang, Byung Joon; Kwon, Seung-Hae; Kee, Yun

    2015-01-01

    Precise 3D spatial mapping of cells and their connections within living tissues is required to fully understand developmental processes and neural activities. Zebrafish embryos are relatively small and optically transparent, making them the vertebrate model of choice for live in vivo imaging. However, embryonic brains cannot be imaged in their entirety by confocal or two-photon microscopy due to limitations in optical range and scanning speed. Here, we use light-sheet fluorescence microscopy to overcome these limitations and image the entire head of live transgenic zebrafish embryos. We simultaneously imaged cranial neurons and blood vessels during embryogenesis, generating comprehensive 3D maps that provide insight into the coordinated morphogenesis of the nervous system and vasculature during early development. In addition, blood cells circulating through the entire head, vagal and cardiac vasculature were also visualized at high resolution in a 3D movie. These data provide the foundation for the construction of a complete 4D atlas of zebrafish embryogenesis and neural activity. PMID:26429501

  6. 3D reconstruction of SEM images by use of optical photogrammetry software.

    PubMed

    Eulitz, Mona; Reiss, Gebhard

    2015-08-01

    Reconstruction of the three-dimensional (3D) surface of an object to be examined is widely used for structure analysis in science and many biological questions require information about their true 3D structure. For Scanning Electron Microscopy (SEM) there has been no efficient non-destructive solution for reconstruction of the surface morphology to date. The well-known method of recording stereo pair images generates a 3D stereoscope reconstruction of a section, but not of the complete sample surface. We present a simple and non-destructive method of 3D surface reconstruction from SEM samples based on the principles of optical close range photogrammetry. In optical close range photogrammetry a series of overlapping photos is used to generate a 3D model of the surface of an object. We adapted this method to the special SEM requirements. Instead of moving a detector around the object, the object itself was rotated. A series of overlapping photos was stitched and converted into a 3D model using the software commonly used for optical photogrammetry. A rabbit kidney glomerulus was used to demonstrate the workflow of this adaption. The reconstruction produced a realistic and high-resolution 3D mesh model of the glomerular surface. The study showed that SEM micrographs are suitable for 3D reconstruction by optical photogrammetry. This new approach is a simple and useful method of 3D surface reconstruction and suitable for various applications in research and teaching.

  7. Flexydos3D: A new deformable anthropomorphic 3D dosimeter readout with optical CT scanning

    NASA Astrophysics Data System (ADS)

    De Deene, Yves; Hill, Robin; Skyt, Peter S.; Booth, Jeremy

    2015-01-01

    A new deformable polydimethylsiloxane (PDMS) based dosimeter is proposed that can be cast in an anthropomorphic shape and that can be used for 3D radiation dosimetry of deformable targets. The new material has additional favorable characteristics as it is tissue equivalent for high-energy photons, easy to make and is non-toxic. In combination with dual wavelength optical scanning, it is a powerful dosimeter for dose verification of image gated or organ tracked radiotherapy with moving and deforming targets.

  8. Optical-CT imaging of complex 3D dose distributions

    NASA Astrophysics Data System (ADS)

    Oldham, Mark; Kim, Leonard; Hugo, Geoffrey

    2005-04-01

    The limitations of conventional dosimeters restrict the comprehensiveness of verification that can be performed for advanced radiation treatments presenting an immediate and substantial problem for clinics attempting to implement these techniques. In essence, the rapid advances in the technology of radiation delivery have not been paralleled by corresponding advances in the ability to verify these treatments. Optical-CT gel-dosimetry is a relatively new technique with potential to address this imbalance by providing high resolution 3D dose maps in polymer and radiochromic gel dosimeters. We have constructed a 1st generation optical-CT scanner capable of high resolution 3D dosimetry and applied it to a number of simple and increasingly complex dose distributions including intensity-modulated-radiation-therapy (IMRT). Prior to application to IMRT, the robustness of optical-CT gel dosimetry was investigated on geometry and variable attenuation phantoms. Physical techniques and image processing methods were developed to minimize deleterious effects of refraction, reflection, and scattered laser light. Here we present results of investigations into achieving accurate high-resolution 3D dosimetry with optical-CT, and show clinical examples of 3D IMRT dosimetry verification. In conclusion, optical-CT gel dosimetry can provide high resolution 3D dose maps that greatly facilitate comprehensive verification of complex 3D radiation treatments. Good agreement was observed at high dose levels (>50%) between planned and measured dose distributions. Some systematic discrepancies were observed however (rms discrepancy 3% at high dose levels) indicating further work is required to eliminate confounding factors presently compromising the accuracy of optical-CT 3D gel-dosimetry.

  9. Fiber optic coherent laser radar 3D vision system

    SciTech Connect

    Clark, R.B.; Gallman, P.G.; Slotwinski, A.R.; Wagner, K.; Weaver, S.; Xu, Jieping

    1996-12-31

    This CLVS will provide a substantial advance in high speed computer vision performance to support robotic Environmental Management (EM) operations. This 3D system employs a compact fiber optic based scanner and operator at a 128 x 128 pixel frame at one frame per second with a range resolution of 1 mm over its 1.5 meter working range. Using acousto-optic deflectors, the scanner is completely randomly addressable. This can provide live 3D monitoring for situations where it is necessary to update once per second. This can be used for decontamination and decommissioning operations in which robotic systems are altering the scene such as in waste removal, surface scarafacing, or equipment disassembly and removal. The fiber- optic coherent laser radar based system is immune to variations in lighting, color, or surface shading, which have plagued the reliability of existing 3D vision systems, while providing substantially superior range resolution.

  10. 3D printed long period gratings for optical fibers.

    PubMed

    Iezzi, Victor Lambin; Boisvert, Jean-Sébastien; Loranger, Sébastien; Kashyap, Raman

    2016-04-15

    We demonstrate a simple technique for implementing long period grating (LPG) structures by the use of a 3D printer. This Letter shows a way of manipulating the mode coupling within an optical fiber by applying stress through an external 3D printed periodic structure. Different LPG lengths and periods have been studied, as well as the effect of the applied stress on the coupling efficiency from the fundamental mode to cladding modes. The technique is very simple, highly flexible, affordable, and easy to implement without the need of altering the optical fiber. This Letter is part of a growing line of interest in the use of 3D printers for optical applications. PMID:27082365

  11. 3D fluorescence anisotropy imaging using selective plane illumination microscopy

    PubMed Central

    Hedde, Per Niklas; Ranjit, Suman; Gratton, Enrico

    2015-01-01

    Fluorescence anisotropy imaging is a popular method to visualize changes in organization and conformation of biomolecules within cells and tissues. In such an experiment, depolarization effects resulting from differences in orientation, proximity and rotational mobility of fluorescently labeled molecules are probed with high spatial resolution. Fluorescence anisotropy is typically imaged using laser scanning and epifluorescence-based approaches. Unfortunately, those techniques are limited in either axial resolution, image acquisition speed, or by photobleaching. In the last decade, however, selective plane illumination microscopy has emerged as the preferred choice for three-dimensional time lapse imaging combining axial sectioning capability with fast, camera-based image acquisition, and minimal light exposure. We demonstrate how selective plane illumination microscopy can be utilized for three-dimensional fluorescence anisotropy imaging of live cells. We further examined the formation of focal adhesions by three-dimensional time lapse anisotropy imaging of CHO-K1 cells expressing an EGFP-paxillin fusion protein. PMID:26368202

  12. Optical fabrication of lightweighted 3D printed mirrors

    NASA Astrophysics Data System (ADS)

    Herzog, Harrison; Segal, Jacob; Smith, Jeremy; Bates, Richard; Calis, Jacob; De La Torre, Alyssa; Kim, Dae Wook; Mici, Joni; Mireles, Jorge; Stubbs, David M.; Wicker, Ryan

    2015-09-01

    Direct Metal Laser Sintering (DMLS) and Electron Beam Melting (EBM) 3D printing technologies were utilized to create lightweight, optical grade mirrors out of AlSi10Mg aluminum and Ti6Al4V titanium alloys at the University of Arizona in Tucson. The mirror prototypes were polished to meet the λ/20 RMS and λ/4 P-V surface figure requirements. The intent of this project was to design topologically optimized mirrors that had a high specific stiffness and low surface displacement. Two models were designed using Altair Inspire software, and the mirrors had to endure the polishing process with the necessary stiffness to eliminate print-through. Mitigating porosity of the 3D printed mirror blanks was a challenge in the face of reconciling new printing technologies with traditional optical polishing methods. The prototypes underwent Hot Isostatic Press (HIP) and heat treatment to improve density, eliminate porosity, and relieve internal stresses. Metal 3D printing allows for nearly unlimited topological constraints on design and virtually eliminates the need for a machine shop when creating an optical quality mirror. This research can lead to an increase in mirror mounting support complexity in the manufacturing of lightweight mirrors and improve overall process efficiency. The project aspired to have many future applications of light weighted 3D printed mirrors, such as spaceflight. This paper covers the design/fab/polish/test of 3D printed mirrors, thermal/structural finite element analysis, and results.

  13. Simultaneous multiplane imaging for 3D confocal microscopy using high-speed z-scanning multiplexing

    NASA Astrophysics Data System (ADS)

    Duocastella, Marti; Vicidomini, Giuseppe; Diaspro, Alberto

    2015-03-01

    One of the key frontiers in optical imaging is to maximize the spatial information retrieved from a sample while minimizing acquisition time. Confocal laser scanning microscopy is a powerful imaging modality that allows real-time and high-resolution acquisition of two-dimensional (2D) sections. However, in order to obtain information from threedimensional (3D) volumes it is currently limited by a stepwise process that consists of acquiring multiple 2D sections from different focal planes by slow z-focus translation. Here, we present a novel method that enables the capture of an entire 3D sample in a single step. Our approach is based on an acoustically-driven varifocal lens integrated in a commercial confocal system that enables axial focus scanning at speeds of 140 kHz or above. Such high-speed allows for one or multiple focus sweeps on a pixel by pixel basis. By using a fast acquisition card, we can assign the photons detected at each pixel to their corresponding focal plane allowing simultaneous multiplane imaging. We exemplify this novel 3D confocal microscopy technique by imaging different biological fluorescent samples and comparing them with those obtained using traditional z-scanners. Based on these results, we find that image quality in this novel approach is similar to that obtained with traditional confocal methods, while speed is only limited by signal-to-noise-ratio. As the sensitivity of photodetectors increases and more efficient fluorescent labeling is developed, this novel 3D method can result in significant reduction in acquisition time allowing the study of new fundamental processes in science.

  14. Optical imaging. Expansion microscopy.

    PubMed

    Chen, Fei; Tillberg, Paul W; Boyden, Edward S

    2015-01-30

    In optical microscopy, fine structural details are resolved by using refraction to magnify images of a specimen. We discovered that by synthesizing a swellable polymer network within a specimen, it can be physically expanded, resulting in physical magnification. By covalently anchoring specific labels located within the specimen directly to the polymer network, labels spaced closer than the optical diffraction limit can be isotropically separated and optically resolved, a process we call expansion microscopy (ExM). Thus, this process can be used to perform scalable superresolution microscopy with diffraction-limited microscopes. We demonstrate ExM with apparent ~70-nanometer lateral resolution in both cultured cells and brain tissue, performing three-color superresolution imaging of ~10(7) cubic micrometers of the mouse hippocampus with a conventional confocal microscope.

  15. Optically Addressed Spatial Light Modulators for 3d Display

    NASA Astrophysics Data System (ADS)

    Collings, N.

    An optically addressed spatial light modulator (OASLM) records the image on a write beam and transfers it to a read beam. Some example application areas are: image transduction; optical correlation; adaptive optics; and optical neural networks. Current interest in OASLMs has been generated by the work of Qinetiq on 3D display. This work is based on Active tiling, where an image can be recorded in one part of the device and is memorised, whilst the remainder of the device is updated with images. This paper will explain this system and survey the technological alternatives for this application.

  16. Registration and 3D visualization of large microscopy images

    NASA Astrophysics Data System (ADS)

    Mosaliganti, Kishore; Pan, Tony; Sharp, Richard; Ridgway, Randall; Iyengar, Srivathsan; Gulacy, Alexandra; Wenzel, Pamela; de Bruin, Alain; Machiraju, Raghu; Huang, Kun; Leone, Gustavo; Saltz, Joel

    2006-03-01

    Inactivation of the retinoblastoma gene in mouse embryos causes tissue infiltrations into critical sections of the placenta, which has been shown to affect fetal survivability. Our collaborators in cancer genetics are extremely interested in examining the three dimensional nature of these infiltrations given a stack of two dimensional light microscopy images. Three sets of wildtype and mutant placentas was sectioned serially and digitized using a commercial light microscopy scanner. Each individual placenta dataset consisted of approximately 1000 images totaling 700 GB in size, which were registered into a volumetric dataset using National Library of Medicine's (NIH/NLM) Insight Segmentation and Registration Toolkit (ITK). This paper describes our method for image registration to aid in volume visualization of tissue level intermixing for both wildtype and Rb - specimens. The registration process faces many challenges arising from the large image sizes, damages during sectioning, staining gradients both within and across sections, and background noise. These issues limit the direct application of standard registration techniques due to frequent convergence to local solutions. In this work, we develop a mixture of automated and semi-automated enhancements with ground-truth validation for the mutual information-based registration algorithm. Our final volume renderings clearly show tissue intermixing differences between both wildtype and Rb - specimens which are not obvious prior to registration.

  17. Automatic respiration tracking for radiotherapy using optical 3D camera

    NASA Astrophysics Data System (ADS)

    Li, Tuotuo; Geng, Jason; Li, Shidong

    2013-03-01

    Rapid optical three-dimensional (O3D) imaging systems provide accurate digitized 3D surface data in real-time, with no patient contact nor radiation. The accurate 3D surface images offer crucial information in image-guided radiation therapy (IGRT) treatments for accurate patient repositioning and respiration management. However, applications of O3D imaging techniques to image-guided radiotherapy have been clinically challenged by body deformation, pathological and anatomical variations among individual patients, extremely high dimensionality of the 3D surface data, and irregular respiration motion. In existing clinical radiation therapy (RT) procedures target displacements are caused by (1) inter-fractional anatomy changes due to weight, swell, food/water intake; (2) intra-fractional variations from anatomy changes within any treatment session due to voluntary/involuntary physiologic processes (e.g. respiration, muscle relaxation); (3) patient setup misalignment in daily reposition due to user errors; and (4) changes of marker or positioning device, etc. Presently, viable solution is lacking for in-vivo tracking of target motion and anatomy changes during the beam-on time without exposing patient with additional ionized radiation or high magnet field. Current O3D-guided radiotherapy systems relay on selected points or areas in the 3D surface to track surface motion. The configuration of the marks or areas may change with time that makes it inconsistent in quantifying and interpreting the respiration patterns. To meet the challenge of performing real-time respiration tracking using O3D imaging technology in IGRT, we propose a new approach to automatic respiration motion analysis based on linear dimensionality reduction technique based on PCA (principle component analysis). Optical 3D image sequence is decomposed with principle component analysis into a limited number of independent (orthogonal) motion patterns (a low dimension eigen-space span by eigen-vectors). New

  18. Virtual 3D microscopy using multiplane whole slide images in diagnostic pathology.

    PubMed

    Kalinski, Thomas; Zwönitzer, Ralf; Sel, Saadettin; Evert, Matthias; Guenther, Thomas; Hofmann, Harald; Bernarding, Johannes; Roessner, Albert

    2008-08-01

    To reproduce focusing in virtual microscopy, it is necessary to construct 3-dimensional (3D) virtual slides composed of whole slide images with different focuses. As focusing is frequently used for the assessment of Helicobacter pylori colonization in diagnostic pathology, we prepared virtual 3D slides with up to 9 focus planes from 144 gastric biopsy specimens with or without H pylori gastritis. The biopsy specimens were diagnosed in a blinded manner by 3 pathologists according to the updated Sydney classification using conventional microscopy, virtual microscopy with a single focus plane, and virtual 3D microscopy with 5 and 9 focus planes enabling virtual focusing. Regarding the classification of H pylori, we found a positive correlation between the number of focus planes used in virtual microscopy and the number of correct diagnoses as determined by conventional microscopy. Concerning H pylori positivity, the specificity and sensitivity of virtual 3D microscopy using virtual slides with 9 focus planes achieved a minimum of 0.95 each, which was approximately the same as in conventional microscopy. We consider virtual 3D microscopy appropriate for primary diagnosis of H pylori gastritis and equivalent to conventional microscopy.

  19. Nanoimprint of a 3D structure on an optical fiber for light wavefront manipulation

    NASA Astrophysics Data System (ADS)

    Calafiore, Giuseppe; Koshelev, Alexander; Allen, Frances I.; Dhuey, Scott; Sassolini, Simone; Wong, Edward; Lum, Paul; Munechika, Keiko; Cabrini, Stefano

    2016-09-01

    Integration of complex photonic structures onto optical fiber facets enables powerful platforms with unprecedented optical functionalities. Conventional nanofabrication technologies, however, do not permit viable integration of complex photonic devices onto optical fibers owing to their low throughput and high cost. In this paper we report the fabrication of a three-dimensional structure achieved by direct nanoimprint lithography on the facet of an optical fiber. Nanoimprint processes and tools were specifically developed to enable a high lithographic accuracy and coaxial alignment of the optical device with respect to the fiber core. To demonstrate the capability of this new approach, a 3D beam splitter has been designed, imprinted and optically characterized. Scanning electron microscopy and optical measurements confirmed the good lithographic capabilities of the proposed approach as well as the desired optical performance of the imprinted structure. The inexpensive solution presented here should enable advancements in areas such as integrated optics and sensing, achieving enhanced portability and versatility of fiber optic components.

  20. Nanoimprint of a 3D structure on an optical fiber for light wavefront manipulation.

    PubMed

    Calafiore, Giuseppe; Koshelev, Alexander; Allen, Frances I; Dhuey, Scott; Sassolini, Simone; Wong, Edward; Lum, Paul; Munechika, Keiko; Cabrini, Stefano

    2016-09-16

    Integration of complex photonic structures onto optical fiber facets enables powerful platforms with unprecedented optical functionalities. Conventional nanofabrication technologies, however, do not permit viable integration of complex photonic devices onto optical fibers owing to their low throughput and high cost. In this paper we report the fabrication of a three-dimensional structure achieved by direct nanoimprint lithography on the facet of an optical fiber. Nanoimprint processes and tools were specifically developed to enable a high lithographic accuracy and coaxial alignment of the optical device with respect to the fiber core. To demonstrate the capability of this new approach, a 3D beam splitter has been designed, imprinted and optically characterized. Scanning electron microscopy and optical measurements confirmed the good lithographic capabilities of the proposed approach as well as the desired optical performance of the imprinted structure. The inexpensive solution presented here should enable advancements in areas such as integrated optics and sensing, achieving enhanced portability and versatility of fiber optic components.

  1. Imaging of human differentiated 3D neural aggregates using light sheet fluorescence microscopy

    PubMed Central

    Gualda, Emilio J.; Simão, Daniel; Pinto, Catarina; Alves, Paula M.; Brito, Catarina

    2014-01-01

    The development of three dimensional (3D) cell cultures represents a big step for the better understanding of cell behavior and disease in a more natural like environment, providing not only single but multiple cell type interactions in a complex 3D matrix, highly resembling physiological conditions. Light sheet fluorescence microscopy (LSFM) is becoming an excellent tool for fast imaging of such 3D biological structures. We demonstrate the potential of this technique for the imaging of human differentiated 3D neural aggregates in fixed and live samples, namely calcium imaging and cell death processes, showing the power of imaging modality compared with traditional microscopy. The combination of light sheet microscopy and 3D neural cultures will open the door to more challenging experiments involving drug testing at large scale as well as a better understanding of relevant biological processes in a more realistic environment. PMID:25161607

  2. Implementation of 3D Optical Scanning Technology for Automotive Applications.

    PubMed

    Kuş, Abdil

    2009-01-01

    Reverse engineering (RE) is a powerful tool for generating a CAD model from the 3D scan data of a physical part that lacks documentation or has changed from the original CAD design of the part. The process of digitizing a part and creating a CAD model from 3D scan data is less time consuming and provides greater accuracy than manually measuring the part and designing the part from scratch in CAD. 3D optical scanning technology is one of the measurement methods which have evolved over the last few years and it is used in a wide range of areas from industrial applications to art and cultural heritage. It is also used extensively in the automotive industry for applications such as part inspections, scanning of tools without CAD definition, scanning the casting for definition of the stock (i.e. the amount of material to be removed from the surface of the castings) model for CAM programs and reverse engineering. In this study two scanning experiments of automotive applications are illustrated. The first one examines the processes from scanning to re-manufacturing the damaged sheet metal cutting die, using a 3D scanning technique and the second study compares the scanned point clouds data to 3D CAD data for inspection purposes. Furthermore, the deviations of the part holes are determined by using different lenses and scanning parameters. PMID:22573995

  3. Implementation of 3D Optical Scanning Technology for Automotive Applications

    PubMed Central

    Kuş, Abdil

    2009-01-01

    Reverse engineering (RE) is a powerful tool for generating a CAD model from the 3D scan data of a physical part that lacks documentation or has changed from the original CAD design of the part. The process of digitizing a part and creating a CAD model from 3D scan data is less time consuming and provides greater accuracy than manually measuring the part and designing the part from scratch in CAD. 3D optical scanning technology is one of the measurement methods which have evolved over the last few years and it is used in a wide range of areas from industrial applications to art and cultural heritage. It is also used extensively in the automotive industry for applications such as part inspections, scanning of tools without CAD definition, scanning the casting for definition of the stock (i.e. the amount of material to be removed from the surface of the castings) model for CAM programs and reverse engineering. In this study two scanning experiments of automotive applications are illustrated. The first one examines the processes from scanning to re-manufacturing the damaged sheet metal cutting die, using a 3D scanning technique and the second study compares the scanned point clouds data to 3D CAD data for inspection purposes. Furthermore, the deviations of the part holes are determined by using different lenses and scanning parameters. PMID:22573995

  4. Implementation of 3D Optical Scanning Technology for Automotive Applications.

    PubMed

    Kuş, Abdil

    2009-01-01

    Reverse engineering (RE) is a powerful tool for generating a CAD model from the 3D scan data of a physical part that lacks documentation or has changed from the original CAD design of the part. The process of digitizing a part and creating a CAD model from 3D scan data is less time consuming and provides greater accuracy than manually measuring the part and designing the part from scratch in CAD. 3D optical scanning technology is one of the measurement methods which have evolved over the last few years and it is used in a wide range of areas from industrial applications to art and cultural heritage. It is also used extensively in the automotive industry for applications such as part inspections, scanning of tools without CAD definition, scanning the casting for definition of the stock (i.e. the amount of material to be removed from the surface of the castings) model for CAM programs and reverse engineering. In this study two scanning experiments of automotive applications are illustrated. The first one examines the processes from scanning to re-manufacturing the damaged sheet metal cutting die, using a 3D scanning technique and the second study compares the scanned point clouds data to 3D CAD data for inspection purposes. Furthermore, the deviations of the part holes are determined by using different lenses and scanning parameters.

  5. Tactile-optical 3D sensor applying image processing

    NASA Astrophysics Data System (ADS)

    Neuschaefer-Rube, Ulrich; Wissmann, Mark

    2009-01-01

    The tactile-optical probe (so-called fiber probe) is a well-known probe in micro-coordinate metrology. It consists of an optical fiber with a probing element at its end. This probing element is adjusted in the imaging plane of the optical system of an optical coordinate measuring machine (CMM). It can be illuminated through the fiber by a LED. The position of the probe is directly detected by image processing algorithms available in every modern optical CMM and not by deflections at the fixation of the probing shaft. Therefore, the probing shaft can be very thin and flexible. This facilitates the measurement with very small probing forces and the realization of very small probing elements (diameter: down to 10 μm). A limitation of this method is that at present the probe does not have full 3D measurement capability. At the Physikalisch-Technische Bundesanstalt (PTB), several arrangements and measurement principles for a full 3D tactile-optical probe have been implemented and tested successfully in cooperation with Werth-Messtechnik, Giessen, Germany. This contribution provides an overview of the results of these activities.

  6. Optical Sensors and Methods for Underwater 3D Reconstruction

    PubMed Central

    Massot-Campos, Miquel; Oliver-Codina, Gabriel

    2015-01-01

    This paper presents a survey on optical sensors and methods for 3D reconstruction in underwater environments. The techniques to obtain range data have been listed and explained, together with the different sensor hardware that makes them possible. The literature has been reviewed, and a classification has been proposed for the existing solutions. New developments, commercial solutions and previous reviews in this topic have also been gathered and considered. PMID:26694389

  7. Nano-spatial parameters from 3D to 2D lattice dimensionality by organic variant in [ZnCl4]- [R]+ hybrid materials: Structure, architecture-lattice dimensionality, microscopy, optical Eg and PL correlations

    NASA Astrophysics Data System (ADS)

    Kumar, Ajit; Verma, Sanjay K.; Alvi, P. A.; Jasrotia, Dinesh

    2016-04-01

    The nanospatial morphological features of [ZnCl]- [C5H4NCH3]+ hybrid derivative depicts 28 nm granular size and 3D spreader shape packing pattern as analyzed by FESEM and single crystal XRD structural studies. The organic moiety connect the inorganic components through N-H+…Cl- hydrogen bond to form a hybrid composite, the replacement of organic derivatives from 2-methylpyridine to 2-Amino-5-choloropyridine results the increase in granular size from 28nm to 60nm and unit cell packing pattern from 3D-2D lattice dimensionality along ac plane. The change in optical energy direct band gap value from 3.01eV for [ZnCl]- [C5H4NCH3]+ (HM1) to 3.42eV for [ZnCl]- [C5H5ClN2]+ (HM2) indicates the role of organic moiety in optical properties of hybrid materials. The photoluminescence emission spectra is observed in the wavelength range of 370 to 600 nm with maximum peak intensity of 9.66a.u. at 438 nm for (HM1) and 370 to 600 nm with max peak intensity of 9.91 a.u. at 442 nm for (HM2), indicating that the emission spectra lies in visible range. PL excitation spectra depicts the maximum excitation intensity [9.8] at 245.5 nm for (HM1) and its value of 9.9 a.u. at 294 nm, specify the excitation spectra lies in UV range. Photoluminescence excitation spectra is observed in the wavelength range of 280 to 350 nm with maximum peak intensity of 9.4 a.u. at 285.5 nm and 9.9 a.u. at 294 and 297 nm, indicating excitation in the UV spectrum. Single crystal growth process and detailed physiochemical characterization such as XRD, FESEM image analysis photoluminescence property reveals the structure stability with non-covalent interactions, lattice dimensionality (3D-2D) correlations interweaving into the design of inorganic-organic hybrid materials.

  8. Open-source 3D-printable optics equipment.

    PubMed

    Zhang, Chenlong; Anzalone, Nicholas C; Faria, Rodrigo P; Pearce, Joshua M

    2013-01-01

    Just as the power of the open-source design paradigm has driven down the cost of software to the point that it is accessible to most people, the rise of open-source hardware is poised to drive down the cost of doing experimental science to expand access to everyone. To assist in this aim, this paper introduces a library of open-source 3-D-printable optics components. This library operates as a flexible, low-cost public-domain tool set for developing both research and teaching optics hardware. First, the use of parametric open-source designs using an open-source computer aided design package is described to customize the optics hardware for any application. Second, details are provided on the use of open-source 3-D printers (additive layer manufacturing) to fabricate the primary mechanical components, which are then combined to construct complex optics-related devices. Third, the use of the open-source electronics prototyping platform are illustrated as control for optical experimental apparatuses. This study demonstrates an open-source optical library, which significantly reduces the costs associated with much optical equipment, while also enabling relatively easily adapted customizable designs. The cost reductions in general are over 97%, with some components representing only 1% of the current commercial investment for optical products of similar function. The results of this study make its clear that this method of scientific hardware development enables a much broader audience to participate in optical experimentation both as research and teaching platforms than previous proprietary methods.

  9. Open-Source 3D-Printable Optics Equipment

    PubMed Central

    Zhang, Chenlong; Anzalone, Nicholas C.; Faria, Rodrigo P.; Pearce, Joshua M.

    2013-01-01

    Just as the power of the open-source design paradigm has driven down the cost of software to the point that it is accessible to most people, the rise of open-source hardware is poised to drive down the cost of doing experimental science to expand access to everyone. To assist in this aim, this paper introduces a library of open-source 3-D-printable optics components. This library operates as a flexible, low-cost public-domain tool set for developing both research and teaching optics hardware. First, the use of parametric open-source designs using an open-source computer aided design package is described to customize the optics hardware for any application. Second, details are provided on the use of open-source 3-D printers (additive layer manufacturing) to fabricate the primary mechanical components, which are then combined to construct complex optics-related devices. Third, the use of the open-source electronics prototyping platform are illustrated as control for optical experimental apparatuses. This study demonstrates an open-source optical library, which significantly reduces the costs associated with much optical equipment, while also enabling relatively easily adapted customizable designs. The cost reductions in general are over 97%, with some components representing only 1% of the current commercial investment for optical products of similar function. The results of this study make its clear that this method of scientific hardware development enables a much broader audience to participate in optical experimentation both as research and teaching platforms than previous proprietary methods. PMID:23544104

  10. Open-source 3D-printable optics equipment.

    PubMed

    Zhang, Chenlong; Anzalone, Nicholas C; Faria, Rodrigo P; Pearce, Joshua M

    2013-01-01

    Just as the power of the open-source design paradigm has driven down the cost of software to the point that it is accessible to most people, the rise of open-source hardware is poised to drive down the cost of doing experimental science to expand access to everyone. To assist in this aim, this paper introduces a library of open-source 3-D-printable optics components. This library operates as a flexible, low-cost public-domain tool set for developing both research and teaching optics hardware. First, the use of parametric open-source designs using an open-source computer aided design package is described to customize the optics hardware for any application. Second, details are provided on the use of open-source 3-D printers (additive layer manufacturing) to fabricate the primary mechanical components, which are then combined to construct complex optics-related devices. Third, the use of the open-source electronics prototyping platform are illustrated as control for optical experimental apparatuses. This study demonstrates an open-source optical library, which significantly reduces the costs associated with much optical equipment, while also enabling relatively easily adapted customizable designs. The cost reductions in general are over 97%, with some components representing only 1% of the current commercial investment for optical products of similar function. The results of this study make its clear that this method of scientific hardware development enables a much broader audience to participate in optical experimentation both as research and teaching platforms than previous proprietary methods. PMID:23544104

  11. Fiber optic coherent laser radar 3d vision system

    SciTech Connect

    Sebastian, R.L.; Clark, R.B.; Simonson, D.L.

    1994-12-31

    Recent advances in fiber optic component technology and digital processing components have enabled the development of a new 3D vision system based upon a fiber optic FMCW coherent laser radar. The approach includes a compact scanner with no moving parts capable of randomly addressing all pixels. The system maintains the immunity to lighting and surface shading conditions which is characteristic of coherent laser radar. The random pixel addressability allows concentration of scanning and processing on the active areas of a scene, as is done by the human eye-brain system.

  12. Large optical 3D MEMS switches in access networks

    NASA Astrophysics Data System (ADS)

    Madamopoulos, Nicholas; Kaman, Volkan; Yuan, Shifu; Jerphagnon, Olivier; Helkey, Roger; Bowers, John E.

    2007-09-01

    Interest is high among residential customers and businesses for advanced, broadband services such as fast Internet access, electronic commerce, video-on-demand, digital broadcasting, teleconferencing and telemedicine. In order to satisfy such growing demand of end-customers, access technologies such as fiber-to-the-home/building (FTTH/B) are increasingly being deployed. Carriers can reduce maintenance costs, minimize technology obsolescence and introduce new services easily by reducing active elements in the fiber access network. However, having a passive optical network (PON) also introduces operational and maintenance challenges. Increased diagnostic monitoring capability of the network becomes a necessity as more and more fibers are provisioned to deliver services to the end-customers. This paper demonstrates the clear advantages that large 3D optical MEMS switches offer in solving these access network problems. The advantages in preventative maintenance, remote monitoring, test and diagnostic capability are highlighted. The low optical insertion loss for all switch optical connections of the switch enables the monitoring, grooming and serving of a large number of PON lines and customers. Furthermore, the 3D MEMS switch is transparent to optical wavelengths and data formats, thus making it easy to incorporate future upgrades, such higher bit rates or DWDM overlay to a PON.

  13. Quantum Degenerate Strontium in a 3D Optical Lattice

    NASA Astrophysics Data System (ADS)

    Aman, J. A.; Desalvo, B. J.; Killian, T. C.

    2014-05-01

    We present our experiments with quantum degenerate neutral strontium in a 3-D optical lattice formed with 532 nm light. Precision control and manipulation of quantum degenerate gases in optical lattices allows for the realization and investigation of tunable many-body systems. Strontium, in particular, has been studied extensively in optical lattices due to the narrow 5s21S0 --> 5 s 5 p3Pj transitions for use as an atomic clock. However, in the present work, we take advantage of these narrow transitions together with strontium's unique isotopic properties to investigate interaction regimes inaccessible to alkali atoms. Among the topics we plan to explore are formation of ultracold molecules using an optical Feshbach resonance as well as the effects of dissipation on atom dynamics. This work was supported by Rice University, Shell, the Welch Foundation (C-1579) and the National Science Foundation (PHY-1205946).

  14. Dual-color 3D superresolution microscopy by combined spectral-demixing and biplane imaging.

    PubMed

    Winterflood, Christian M; Platonova, Evgenia; Albrecht, David; Ewers, Helge

    2015-07-01

    Multicolor three-dimensional (3D) superresolution techniques allow important insight into the relative organization of cellular structures. While a number of innovative solutions have emerged, multicolor 3D techniques still face significant technical challenges. In this Letter we provide a straightforward approach to single-molecule localization microscopy imaging in three dimensions and two colors. We combine biplane imaging and spectral-demixing, which eliminates a number of problems, including color cross-talk, chromatic aberration effects, and problems with color registration. We present 3D dual-color images of nanoscopic structures in hippocampal neurons with a 3D compound resolution routinely achieved only in a single color.

  15. Constructing 3D microtubule networks using holographic optical trapping

    PubMed Central

    Bergman, J.; Osunbayo, O.; Vershinin, M.

    2015-01-01

    Developing abilities to assemble nanoscale structures is a major scientific and engineering challenge. We report a technique which allows precise positioning and manipulation of individual rigid filaments, enabling construction of custom-designed 3D filament networks. This approach uses holographic optical trapping (HOT) for nano-positioning and microtubules (MTs) as network building blocks. MTs are desirable engineering components due to their high aspect ratio, rigidity, and their ability to serve as substrate for directed nano-transport, reflecting their roles in the eukaryotic cytoskeleton. The 3D architecture of MT cytoskeleton is a significant component of its function, however experimental tools to study the roles of this geometric complexity in a controlled environment have been lacking. We demonstrate the broad capabilities of our system by building a self-supporting 3D MT-based nanostructure and by conducting a MT-based transport experiment on a dynamically adjustable 3D MT intersection. Our methodology not only will advance studies of cytoskeletal networks (and associated processes such as MT-based transport) but will also likely find use in engineering nanostructures and devices. PMID:26657337

  16. 3D Human cartilage surface characterization by optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Brill, Nicolai; Riedel, Jörn; Schmitt, Robert; Tingart, Markus; Truhn, Daniel; Pufe, Thomas; Jahr, Holger; Nebelung, Sven

    2015-10-01

    Early diagnosis and treatment of cartilage degeneration is of high clinical interest. Loss of surface integrity is considered one of the earliest and most reliable signs of degeneration, but cannot currently be evaluated objectively. Optical Coherence Tomography (OCT) is an arthroscopically available light-based non-destructive real-time imaging technology that allows imaging at micrometre resolutions to millimetre depths. As OCT-based surface evaluation standards remain to be defined, the present study investigated the diagnostic potential of 3D surface profile parameters in the comprehensive evaluation of cartilage degeneration. To this end, 45 cartilage samples of different degenerative grades were obtained from total knee replacements (2 males, 10 females; mean age 63.8 years), cut to standard size and imaged using a spectral-domain OCT device (Thorlabs, Germany). 3D OCT datasets of 8  ×  8, 4  ×  4 and 1  ×  1 mm (width  ×  length) were obtained and pre-processed (image adjustments, morphological filtering). Subsequent automated surface identification algorithms were used to obtain the 3D primary profiles, which were then filtered and processed using established algorithms employing ISO standards. The 3D surface profile thus obtained was used to calculate a set of 21 3D surface profile parameters, i.e. height (e.g. Sa), functional (e.g. Sk), hybrid (e.g. Sdq) and segmentation-related parameters (e.g. Spd). Samples underwent reference histological assessment according to the Degenerative Joint Disease classification. Statistical analyses included calculation of Spearman’s rho and assessment of inter-group differences using the Kruskal Wallis test. Overall, the majority of 3D surface profile parameters revealed significant degeneration-dependent differences and correlations with the exception of severe end-stage degeneration and were of distinct diagnostic value in the assessment of surface integrity. None of the 3D

  17. Multimodal 3D cancer-mimicking optical phantom

    PubMed Central

    Smith, Gennifer T.; Lurie, Kristen L.; Zlatev, Dimitar V.; Liao, Joseph C.; Ellerbee Bowden, Audrey K.

    2016-01-01

    Three-dimensional (3D) organ-mimicking phantoms provide realistic imaging environments for testing various aspects of optical systems, including for evaluating new probe designs, characterizing the diagnostic potential of new technologies, and assessing novel image processing algorithms prior to validation in real tissue. We introduce and characterize the use of a new material, Dragon Skin (Smooth-On Inc.), and fabrication technique, air-brushing, for fabrication of a 3D phantom that mimics the appearance of a real organ under multiple imaging modalities. We demonstrate the utility of the material and technique by fabricating the first 3D, hollow bladder phantom with realistic normal and multi-stage pathology features suitable for endoscopic detection using the gold standard imaging technique, white light cystoscopy (WLC), as well as the complementary imaging modalities of optical coherence tomography and blue light cystoscopy, which are aimed at improving the sensitivity and specificity of WLC to bladder cancer detection. The flexibility of the material and technique used for phantom construction allowed for the representation of a wide range of diseased tissue states, ranging from inflammation (benign) to high-grade cancerous lesions. Such phantoms can serve as important tools for trainee education and evaluation of new endoscopic instrumentation. PMID:26977369

  18. Wave optics theory and 3-D deconvolution for the light field microscope

    PubMed Central

    Broxton, Michael; Grosenick, Logan; Yang, Samuel; Cohen, Noy; Andalman, Aaron; Deisseroth, Karl; Levoy, Marc

    2013-01-01

    Light field microscopy is a new technique for high-speed volumetric imaging of weakly scattering or fluorescent specimens. It employs an array of microlenses to trade off spatial resolution against angular resolution, thereby allowing a 4-D light field to be captured using a single photographic exposure without the need for scanning. The recorded light field can then be used to computationally reconstruct a full volume. In this paper, we present an optical model for light field microscopy based on wave optics, instead of previously reported ray optics models. We also present a 3-D deconvolution method for light field microscopy that is able to reconstruct volumes at higher spatial resolution, and with better optical sectioning, than previously reported. To accomplish this, we take advantage of the dense spatio-angular sampling provided by a microlens array at axial positions away from the native object plane. This dense sampling permits us to decode aliasing present in the light field to reconstruct high-frequency information. We formulate our method as an inverse problem for reconstructing the 3-D volume, which we solve using a GPU-accelerated iterative algorithm. Theoretical limits on the depth-dependent lateral resolution of the reconstructed volumes are derived. We show that these limits are in good agreement with experimental results on a standard USAF 1951 resolution target. Finally, we present 3-D reconstructions of pollen grains that demonstrate the improvements in fidelity made possible by our method. PMID:24150383

  19. Optic flow aided navigation and 3D scene reconstruction

    NASA Astrophysics Data System (ADS)

    Rollason, Malcolm

    2013-10-01

    An important enabler for low cost airborne systems is the ability to exploit low cost inertial instruments. An Inertial Navigation System (INS) can provide a navigation solution, when GPS is denied, by integrating measurements from inertial sensors. However, the gyrometer and accelerometer biases of low cost inertial sensors cause compound errors in the integrated navigation solution. This paper describes experiments to establish whether (and to what extent) the navigation solution can be aided by fusing measurements from an on-board video camera with measurements from the inertial sensors. The primary aim of the work was to establish whether optic flow aided navigation is beneficial even when the 3D structure within the observed scene is unknown. A further aim was to investigate whether an INS can help to infer 3D scene content from video. Experiments with both real and synthetic data have been conducted. Real data was collected using an AR Parrot quadrotor. Empirical results illustrate that optic flow provides a useful aid to navigation even when the 3D structure of the observed scene is not known. With optic flow aiding of the INS, the computed trajectory is consistent with the true camera motion, whereas the unaided INS yields a rapidly increasing position error (the data represents ~40 seconds, after which the unaided INS is ~50 metres in error and has passed through the ground). The results of the Monte Carlo simulation concur with the empirical result. Position errors, which grow as a quadratic function of time when unaided, are substantially checked by the availability of optic flow measurements.

  20. Beam Optics Analysis - An Advanced 3D Trajectory Code

    SciTech Connect

    Ives, R. Lawrence; Bui, Thuc; Vogler, William; Neilson, Jeff; Read, Mike; Shephard, Mark; Bauer, Andrew; Datta, Dibyendu; Beal, Mark

    2006-01-03

    Calabazas Creek Research, Inc. has completed initial development of an advanced, 3D program for modeling electron trajectories in electromagnetic fields. The code is being used to design complex guns and collectors. Beam Optics Analysis (BOA) is a fully relativistic, charged particle code using adaptive, finite element meshing. Geometrical input is imported from CAD programs generating ACIS-formatted files. Parametric data is inputted using an intuitive, graphical user interface (GUI), which also provides control of convergence, accuracy, and post processing. The program includes a magnetic field solver, and magnetic information can be imported from Maxwell 2D/3D and other programs. The program supports thermionic emission and injected beams. Secondary electron emission is also supported, including multiple generations. Work on field emission is in progress as well as implementation of computer optimization of both the geometry and operating parameters. The principle features of the program and its capabilities are presented.

  1. Beam Optics Analysis — An Advanced 3D Trajectory Code

    NASA Astrophysics Data System (ADS)

    Ives, R. Lawrence; Bui, Thuc; Vogler, William; Neilson, Jeff; Read, Mike; Shephard, Mark; Bauer, Andrew; Datta, Dibyendu; Beal, Mark

    2006-01-01

    Calabazas Creek Research, Inc. has completed initial development of an advanced, 3D program for modeling electron trajectories in electromagnetic fields. The code is being used to design complex guns and collectors. Beam Optics Analysis (BOA) is a fully relativistic, charged particle code using adaptive, finite element meshing. Geometrical input is imported from CAD programs generating ACIS-formatted files. Parametric data is inputted using an intuitive, graphical user interface (GUI), which also provides control of convergence, accuracy, and post processing. The program includes a magnetic field solver, and magnetic information can be imported from Maxwell 2D/3D and other programs. The program supports thermionic emission and injected beams. Secondary electron emission is also supported, including multiple generations. Work on field emission is in progress as well as implementation of computer optimization of both the geometry and operating parameters. The principle features of the program and its capabilities are presented.

  2. Single 3D cell segmentation from optical CT microscope images

    NASA Astrophysics Data System (ADS)

    Xie, Yiting; Reeves, Anthony P.

    2014-03-01

    The automated segmentation of the nucleus and cytoplasm regions in 3D optical CT microscope images has been achieved with two methods, a global threshold gradient based approach and a graph-cut approach. For the first method, the first two peaks of a gradient figure of merit curve are selected as the thresholds for cytoplasm and nucleus segmentation. The second method applies a graph-cut segmentation twice: the first identifies the nucleus region and the second identifies the cytoplasm region. Image segmentation of single cells is important for automated disease diagnostic systems. The segmentation methods were evaluated with 200 3D images consisting of 40 samples of 5 different cell types. The cell types consisted of columnar, macrophage, metaplastic and squamous human cells and cultured A549 cancer cells. The segmented cells were compared with both 2D and 3D reference images and the quality of segmentation was determined by the Dice Similarity Coefficient (DSC). In general, the graph-cut method had a superior performance to the gradient-based method. The graph-cut method achieved an average DSC of 86% and 72% for nucleus and cytoplasm segmentations respectively for the 2D reference images and 83% and 75% for the 3D reference images. The gradient method achieved an average DSC of 72% and 51% for nucleus and cytoplasm segmentation for the 2D reference images and 71% and 51% for the 3D reference images. The DSC of cytoplasm segmentation was significantly lower than for the nucleus since the cytoplasm was not differentiated as well by image intensity from the background.

  3. Multimodal Nonlinear Optical Microscopy

    PubMed Central

    Yue, Shuhua; Slipchenko, Mikhail N.; Cheng, Ji-Xin

    2013-01-01

    Because each nonlinear optical (NLO) imaging modality is sensitive to specific molecules or structures, multimodal NLO imaging capitalizes the potential of NLO microscopy for studies of complex biological tissues. The coupling of multiphoton fluorescence, second harmonic generation, and coherent anti-Stokes Raman scattering (CARS) has allowed investigation of a broad range of biological questions concerning lipid metabolism, cancer development, cardiovascular disease, and skin biology. Moreover, recent research shows the great potential of using CARS microscope as a platform to develop more advanced NLO modalities such as electronic-resonance-enhanced four-wave mixing, stimulated Raman scattering, and pump-probe microscopy. This article reviews the various approaches developed for realization of multimodal NLO imaging as well as developments of new NLO modalities on a CARS microscope. Applications to various aspects of biological and biomedical research are discussed. PMID:24353747

  4. Innovations in 3D printing: a 3D overview from optics to organs.

    PubMed

    Schubert, Carl; van Langeveld, Mark C; Donoso, Larry A

    2014-02-01

    3D printing is a method of manufacturing in which materials, such as plastic or metal, are deposited onto one another in layers to produce a three dimensional object, such as a pair of eye glasses or other 3D objects. This process contrasts with traditional ink-based printers which produce a two dimensional object (ink on paper). To date, 3D printing has primarily been used in engineering to create engineering prototypes. However, recent advances in printing materials have now enabled 3D printers to make objects that are comparable with traditionally manufactured items. In contrast with conventional printers, 3D printing has the potential to enable mass customisation of goods on a large scale and has relevance in medicine including ophthalmology. 3D printing has already been proved viable in several medical applications including the manufacture of eyeglasses, custom prosthetic devices and dental implants. In this review, we discuss the potential for 3D printing to revolutionise manufacturing in the same way as the printing press revolutionised conventional printing. The applications and limitations of 3D printing are discussed; the production process is demonstrated by producing a set of eyeglass frames from 3D blueprints. PMID:24288392

  5. Innovations in 3D printing: a 3D overview from optics to organs.

    PubMed

    Schubert, Carl; van Langeveld, Mark C; Donoso, Larry A

    2014-02-01

    3D printing is a method of manufacturing in which materials, such as plastic or metal, are deposited onto one another in layers to produce a three dimensional object, such as a pair of eye glasses or other 3D objects. This process contrasts with traditional ink-based printers which produce a two dimensional object (ink on paper). To date, 3D printing has primarily been used in engineering to create engineering prototypes. However, recent advances in printing materials have now enabled 3D printers to make objects that are comparable with traditionally manufactured items. In contrast with conventional printers, 3D printing has the potential to enable mass customisation of goods on a large scale and has relevance in medicine including ophthalmology. 3D printing has already been proved viable in several medical applications including the manufacture of eyeglasses, custom prosthetic devices and dental implants. In this review, we discuss the potential for 3D printing to revolutionise manufacturing in the same way as the printing press revolutionised conventional printing. The applications and limitations of 3D printing are discussed; the production process is demonstrated by producing a set of eyeglass frames from 3D blueprints.

  6. Optical monitoring of scoliosis by 3D medical laser scanner

    NASA Astrophysics Data System (ADS)

    Rodríguez-Quiñonez, Julio C.; Sergiyenko, Oleg Yu.; Preciado, Luis C. Basaca; Tyrsa, Vera V.; Gurko, Alexander G.; Podrygalo, Mikhail A.; Lopez, Moises Rivas; Balbuena, Daniel Hernandez

    2014-03-01

    Three dimensional recording of the human body surface or anatomical areas have gained importance in many medical applications. In this paper, our 3D Medical Laser Scanner is presented. It is based on the novel principle of dynamic triangulation. We analyze the method of operation, medical applications, orthopedically diseases as Scoliosis and the most common types of skin to employ the system the most proper way. It is analyzed a group of medical problems related to the application of optical scanning in optimal way. Finally, experiments are conducted to verify the performance of the proposed system and its method uncertainty.

  7. Towards 3-D laser nano patterning in polymer optical materials

    NASA Astrophysics Data System (ADS)

    Scully, Patricia J.; Perrie, Walter

    2015-03-01

    Progress towards 3-D subsurface structuring of polymers using femtosecond lasers is presented. Highly localised refractive index changes can be generated deep in transparent optical polymers without pre doping for photosensitisation or post processing by annealing. Understanding the writing conditions surpasses the limitations of materials, dimensions and chemistry, to facilitate unique structures entirely formed by laser-polymeric interactions to overcome materials, dimensional, refractive index and wavelength constraints.. Numerical aperture, fluence, temporal pulselength, wavelength and incident polarisation are important parameters to be considered, in achieving the desired inscription. Non-linear aspects of multiphoton absorption, plasma generation, filamentation and effects of incident polarisation on the writing conditions will be presented.

  8. 3D refractive index measurements of special optical fibers

    NASA Astrophysics Data System (ADS)

    Yan, Cheng; Huang, Su-Juan; Miao, Zhuang; Chang, Zheng; Zeng, Jun-Zhang; Wang, Ting-Yun

    2016-09-01

    A digital holographic microscopic chromatography-based approach with considerably improved accuracy, simplified configuration and performance stability is proposed to measure three dimensional refractive index of special optical fibers. Based on the approach, a measurement system is established incorporating a modified Mach-Zehnder interferometer and lab-developed supporting software for data processing. In the system, a phase projection distribution of an optical fiber is utilized to obtain an optimal digital hologram recorded by a CCD, and then an angular spectrum theory-based algorithm is adopted to extract the phase distribution information of an object wave. The rotation of the optic fiber enables the experimental measurements of multi-angle phase information. Based on the filtered back projection algorithm, a 3D refraction index of the optical fiber is thus obtained at high accuracy. To evaluate the proposed approach, both PANDA fibers and special elliptical optical fiber are considered in the system. The results measured in PANDA fibers agree well with those measured using S14 Refractive Index Profiler, which is, however, not suitable for measuring the property of a special elliptical fiber.

  9. Digital holographic microscopy for imaging growth and treatment response in 3D tumor models

    NASA Astrophysics Data System (ADS)

    Li, Yuyu; Petrovic, Ljubica; Celli, Jonathan P.; Yelleswarapu, Chandra S.

    2014-03-01

    While three-dimensional tumor models have emerged as valuable tools in cancer research, the ability to longitudinally visualize the 3D tumor architecture restored by these systems is limited with microscopy techniques that provide only qualitative insight into sample depth, or which require terminal fixation for depth-resolved 3D imaging. Here we report the use of digital holographic microscopy (DHM) as a viable microscopy approach for quantitative, non-destructive longitudinal imaging of in vitro 3D tumor models. Following established methods we prepared 3D cultures of pancreatic cancer cells in overlay geometry on extracellular matrix beds and obtained digital holograms at multiple timepoints throughout the duration of growth. The holograms were digitally processed and the unwrapped phase images were obtained to quantify nodule thickness over time under normal growth, and in cultures subject to chemotherapy treatment. In this manner total nodule volumes are rapidly estimated and demonstrated here to show contrasting time dependent changes during growth and in response to treatment. This work suggests the utility of DHM to quantify changes in 3D structure over time and suggests the further development of this approach for time-lapse monitoring of 3D morphological changes during growth and in response to treatment that would otherwise be impractical to visualize.

  10. Rapid prototyping with optical 3D measurement systems

    NASA Astrophysics Data System (ADS)

    Gaessler, J.; Blount, G. N.; Jones, R. M.

    1994-11-01

    One of the important tools for speeding up the prototyping of an new industrial or consumer product is the rapid generation of CAD data from hand-made styling models and moulds. We present a new optical 3D digitizing system which produces in a fully automatic way non- ambiguous, absolute and complete surface coordinate data of very complex objects in a short time. The system named `OptoShape' is based on a projection of sinusoidal fringes with a true grey-level matrix projector. The system measures both non-ambiguous and absolute XYZ surface data with a pronounced robustness towards optical surface properties. By moving the 3D sensor head around the object to be digitized with a 3/5 axes manipulator, multiple range images are obtained and automatically merged into a unified cloud of point coordinates. This set of surface coordinates are transferred to a software package where interactive manipulation, sectioning and semi-automatic generation of CAD surface descriptions are performed. CNC data can also be directly generated from the point surface coordinate data set.

  11. Heads-up 3D Microscopy: An Ergonomic and Educational Approach to Microsurgery.

    PubMed

    Mendez, Bernardino M; Chiodo, Michael V; Vandevender, Darl; Patel, Parit A

    2016-05-01

    Traditional microsurgery can lead surgeons to use postures that cause musculoskeletal fatigue, leaving them more prone to work-related injuries. A new technology from TrueVision transmits the microscopic image onto a 3-dimensional (3D) monitor, allowing surgeons to operate while sitting/standing in a heads-up position. The purpose of this study was to evaluate the feasibility of performing heads-up 3D microscopy as a more ergonomic alternative to traditional microsurgery. A feasibility study was conducted comparing heads-up 3D microscopy and traditional microscopy by performing femoral artery anastomoses on 8 Sprague-Dawley rats. Operative times and patency rates for each technology were compared. The 8 microsurgeons completed a questionnaire comparing image quality, comfort, technical feasibility, and educational value of the 2 technologies. Rat femoral artery anastomoses were successfully carried out by all 8 microsurgeons with each technology. There was no significant difference in anastomosis time between heads-up 3D and traditional microscopy (average times, 34.5 and 33.8 minutes, respectively; P = 0.66). Heads-up 3D microscopy was rated superior in neck and back comfort by 75% of participants. Image resolution, field of view, and technical feasibility were found to be superior or equivalent in 75% of participants, whereas 63% evaluated depth perception to be superior or equivalent. Heads-up 3D microscopy is a new technology that improves comfort for the microsurgeon without compromising image quality or technical feasibility. Its use has become prevalent in the field of ophthalmology and may also have utility in plastic and reconstructive surgery. PMID:27579241

  12. Heads-up 3D Microscopy: An Ergonomic and Educational Approach to Microsurgery.

    PubMed

    Mendez, Bernardino M; Chiodo, Michael V; Vandevender, Darl; Patel, Parit A

    2016-05-01

    Traditional microsurgery can lead surgeons to use postures that cause musculoskeletal fatigue, leaving them more prone to work-related injuries. A new technology from TrueVision transmits the microscopic image onto a 3-dimensional (3D) monitor, allowing surgeons to operate while sitting/standing in a heads-up position. The purpose of this study was to evaluate the feasibility of performing heads-up 3D microscopy as a more ergonomic alternative to traditional microsurgery. A feasibility study was conducted comparing heads-up 3D microscopy and traditional microscopy by performing femoral artery anastomoses on 8 Sprague-Dawley rats. Operative times and patency rates for each technology were compared. The 8 microsurgeons completed a questionnaire comparing image quality, comfort, technical feasibility, and educational value of the 2 technologies. Rat femoral artery anastomoses were successfully carried out by all 8 microsurgeons with each technology. There was no significant difference in anastomosis time between heads-up 3D and traditional microscopy (average times, 34.5 and 33.8 minutes, respectively; P = 0.66). Heads-up 3D microscopy was rated superior in neck and back comfort by 75% of participants. Image resolution, field of view, and technical feasibility were found to be superior or equivalent in 75% of participants, whereas 63% evaluated depth perception to be superior or equivalent. Heads-up 3D microscopy is a new technology that improves comfort for the microsurgeon without compromising image quality or technical feasibility. Its use has become prevalent in the field of ophthalmology and may also have utility in plastic and reconstructive surgery.

  13. Combined scanning probe nanotomography and optical microspectroscopy: a correlative technique for 3D characterization of nanomaterials.

    PubMed

    Mochalov, Konstantin E; Efimov, Anton E; Bobrovsky, Alexey; Agapov, Igor I; Chistyakov, Anton A; Oleinikov, Vladimir; Sukhanova, Alyona; Nabiev, Igor

    2013-10-22

    Combination of 3D structural analysis with optical characterization of the same sample area on the nanoscale is a highly demanded approach in nanophotonics, materials science, and quality control of nanomaterial. We have developed a correlative microscopy technique where the 3D structure of the sample is reconstructed on the nanoscale by means of a "slice-and-view" combination of ultramicrotomy and scanning probe microscopy (scanning probe nanotomography, SPNT), and its optical characteristics are analyzed using microspectroscopy. This approach has been used to determine the direct quantitative relationship of the 3D structural characteristics of nanovolumes of materials with their microscopic optical properties. This technique has been applied to 3D structural and optical characterization of a hybrid material consisting of cholesteric liquid crystals doped with fluorescent quantum dots (QDs) that can be used for photochemical patterning and image recording through the changes in the dissymmetry factor of the circular polarization of QD emission. The differences in the polarization images and fluorescent spectra of this hybrid material have proved to be correlated with the arrangement of the areas of homogeneous distribution and heterogeneous clustering of QDs. The reconstruction of the 3D nanostructure of the liquid crystal matrix in the areas of homogeneous QDs distribution has shown that QDs do not perturb the periodic planar texture of the cholesteric liquid crystal matrix, whereas QD clusters do perturb it. The combined microspectroscopy-nanotomography technique will be important for evaluating the effects of nanoparticles on the structural organization of organic and liquid crystal matrices and biomedical materials, as well as quality control of nanotechnology fabrication processes and products.

  14. Combined scanning probe nanotomography and optical microspectroscopy: a correlative technique for 3D characterization of nanomaterials.

    PubMed

    Mochalov, Konstantin E; Efimov, Anton E; Bobrovsky, Alexey; Agapov, Igor I; Chistyakov, Anton A; Oleinikov, Vladimir; Sukhanova, Alyona; Nabiev, Igor

    2013-10-22

    Combination of 3D structural analysis with optical characterization of the same sample area on the nanoscale is a highly demanded approach in nanophotonics, materials science, and quality control of nanomaterial. We have developed a correlative microscopy technique where the 3D structure of the sample is reconstructed on the nanoscale by means of a "slice-and-view" combination of ultramicrotomy and scanning probe microscopy (scanning probe nanotomography, SPNT), and its optical characteristics are analyzed using microspectroscopy. This approach has been used to determine the direct quantitative relationship of the 3D structural characteristics of nanovolumes of materials with their microscopic optical properties. This technique has been applied to 3D structural and optical characterization of a hybrid material consisting of cholesteric liquid crystals doped with fluorescent quantum dots (QDs) that can be used for photochemical patterning and image recording through the changes in the dissymmetry factor of the circular polarization of QD emission. The differences in the polarization images and fluorescent spectra of this hybrid material have proved to be correlated with the arrangement of the areas of homogeneous distribution and heterogeneous clustering of QDs. The reconstruction of the 3D nanostructure of the liquid crystal matrix in the areas of homogeneous QDs distribution has shown that QDs do not perturb the periodic planar texture of the cholesteric liquid crystal matrix, whereas QD clusters do perturb it. The combined microspectroscopy-nanotomography technique will be important for evaluating the effects of nanoparticles on the structural organization of organic and liquid crystal matrices and biomedical materials, as well as quality control of nanotechnology fabrication processes and products. PMID:23991901

  15. Quantitative 3D Optical Imaging: Applications in Dosimetry and Biophysics

    NASA Astrophysics Data System (ADS)

    Thomas, Andrew Stephen

    Optical-CT has been shown to be a potentially useful imaging tool for the two very different spheres of biologists and radiation therapy physicists, but it has yet to live up to that potential. In radiation therapy, researchers have used optical-CT for the readout of 3D dosimeters, but it is yet to be a clinically relevant tool as the technology is too slow to be considered practical. Biologists have used the technique for structural imaging, but have struggled with emission tomography as the reality of photon attenuation for both excitation and emission have made the images quantitatively irrelevant. Dosimetry. The DLOS (Duke Large field of view Optical-CT Scanner) was designed and constructed to make 3D dosimetry utilizing optical-CT a fast and practical tool while maintaining the accuracy of readout of the previous, slower readout technologies. Upon construction/optimization/implementation of several components including a diffuser, band pass filter, registration mount & fluid filtration system the dosimetry system provides high quality data comparable to or exceeding that of commercial products. In addition, a stray light correction algorithm was tested and implemented. The DLOS in combination with the 3D dosimeter it was designed for, PREAGETM, then underwent rigorous commissioning and benchmarking tests validating its performance against gold standard data including a set of 6 irradiations. DLOS commissioning tests resulted in sub-mm isotropic spatial resolution (MTF >0.5 for frequencies of 1.5lp/mm) and a dynamic range of ˜60dB. Flood field uniformity was 10% and stable after 45minutes. Stray light proved to be small, due to telecentricity, but even the residual can be removed through deconvolution. Benchmarking tests showed the mean 3D passing gamma rate (3%, 3mm, 5% dose threshold) over the 6 benchmark data sets was 97.3% +/- 0.6% (range 96%-98%) scans totaling ˜10 minutes, indicating excellent ability to perform 3D dosimetry while improving the speed of

  16. Optically Trapped Surface-Enhanced Raman Probes Prepared by Silver Photoreduction to 3D Microstructures.

    PubMed

    Vizsnyiczai, Gaszton; Lestyán, Tamás; Joniova, Jaroslava; Aekbote, Badri L; Strejčková, Alena; Ormos, Pál; Miskovsky, Pavol; Kelemen, Lóránd; Bánó, Gregor

    2015-09-15

    3D microstructures partially covered by silver nanoparticles have been developed and tested for surface-enhanced Raman spectroscopy (SERS) in combination with optical tweezers. The microstructures made by two-photon polymerization of SU-8 photoresist were manipulated in a dual beam optical trap. The active area of the structures was covered by a SERS-active silver layer using chemically assisted photoreduction from silver nitrate solutions. Silver layers of different grain size distributions were created by changing the photoreduction parameters and characterized by scanning electron microscopy. The structures were tested by measuring the SERS spectra of emodin and hypericin.

  17. Ultracold polar molecules in a 3D optical lattice

    NASA Astrophysics Data System (ADS)

    Yan, Bo

    2015-05-01

    Ultracold polar molecules, with their long-range electric dipolar interactions, offer new opportunities for studying quantum magnetism and many-body physics. KRb molecules loaded into a three-dimensional (3D) optical lattice allow one to study such a spin-lattice system in a stable environment without losses arising from chemical reactions. In the case with strong lattice confinement along two directions and a weak lattice potential along the third, we find the loss rate is suppressed by the quantum Zeno effect. In a deep 3D lattice with no tunneling, we observe evidences for spin exchange interactions. We use Ramsey spectroscopy to investigate the spin dynamics. By choosing the appropriate lattice polarizations and implementing a spin echo sequence, the single particle dephasing is largely suppressed, leaving the dipolar exchange interactions as the dominant contribution to the observed dynamics. This is supported by many-body theoretical calculations. While this initial demonstration was done with low lattice fillings, our current experimental efforts are focused on increasing the lattice filling fraction. This will greatly benefit the study of complex many-body dynamics with long-range interactions, such as transport of excitations in an out-of-equilibrium system and spin-orbit coupling in a lattice.

  18. New 3D optical digitizer for orthodontic applications

    NASA Astrophysics Data System (ADS)

    Canal Bienzobas, Fernando; Dios, Federico; Garcia-Mateos, Jorge; Rivera, Alejandro

    2002-05-01

    A new three-dimensional optical digitizer working with white light and high accuracy has been developed for applications in orthodontic diagnosis, to store and analyze data taken from plaster dental casts. The scanner consists basically of a triangulation device made of a CCD camera and an active light projector using Gray code and the phase stepping method to sample the surfaces under tests. Dense clouds of 3D points of the object's surface, related to the device's reference system, are generated, from which very exact measurements can be taken. This technique allows us to greatly improve the accuracy of results that previously were obtained by using mainly manual methods. Our system has been tested successfully on many different samples.

  19. Volumetric (3D) compressive sensing spectral domain optical coherence tomography

    PubMed Central

    Xu, Daguang; Huang, Yong; Kang, Jin U.

    2014-01-01

    In this work, we proposed a novel three-dimensional compressive sensing (CS) approach for spectral domain optical coherence tomography (SD OCT) volumetric image acquisition and reconstruction. Instead of taking a spectral volume whose size is the same as that of the volumetric image, our method uses a sub set of the original spectral volume that is under-sampled in all three dimensions, which reduces the amount of spectral measurements to less than 20% of that required by the Shan-non/Nyquist theory. The 3D image is recovered from the under-sampled spectral data dimension-by-dimension using the proposed three-step CS reconstruction strategy. Experimental results show that our method can significantly reduce the sampling rate required for a volumetric SD OCT image while preserving the image quality. PMID:25426320

  20. Volumetric (3D) compressive sensing spectral domain optical coherence tomography.

    PubMed

    Xu, Daguang; Huang, Yong; Kang, Jin U

    2014-11-01

    In this work, we proposed a novel three-dimensional compressive sensing (CS) approach for spectral domain optical coherence tomography (SD OCT) volumetric image acquisition and reconstruction. Instead of taking a spectral volume whose size is the same as that of the volumetric image, our method uses a sub set of the original spectral volume that is under-sampled in all three dimensions, which reduces the amount of spectral measurements to less than 20% of that required by the Shan-non/Nyquist theory. The 3D image is recovered from the under-sampled spectral data dimension-by-dimension using the proposed three-step CS reconstruction strategy. Experimental results show that our method can significantly reduce the sampling rate required for a volumetric SD OCT image while preserving the image quality.

  1. 3D image reconstruction algorithms for cryo-electron-microscopy images of virus particles

    NASA Astrophysics Data System (ADS)

    Doerschuk, Peter C.; Johnson, John E.

    2000-11-01

    A statistical model for the object and the complete image formation process in cryo electron microscopy of viruses is presented. Using this model, maximum likelihood reconstructions of the 3D structure of viruses are computed using the expectation maximization algorithm and an example based on Cowpea mosaic virus is provided.

  2. Nanoparticle imaging. 3D structure of individual nanocrystals in solution by electron microscopy.

    PubMed

    Park, Jungwon; Elmlund, Hans; Ercius, Peter; Yuk, Jong Min; Limmer, David T; Chen, Qian; Kim, Kwanpyo; Han, Sang Hoon; Weitz, David A; Zettl, A; Alivisatos, A Paul

    2015-07-17

    Knowledge about the synthesis, growth mechanisms, and physical properties of colloidal nanoparticles has been limited by technical impediments. We introduce a method for determining three-dimensional (3D) structures of individual nanoparticles in solution. We combine a graphene liquid cell, high-resolution transmission electron microscopy, a direct electron detector, and an algorithm for single-particle 3D reconstruction originally developed for analysis of biological molecules. This method yielded two 3D structures of individual platinum nanocrystals at near-atomic resolution. Because our method derives the 3D structure from images of individual nanoparticles rotating freely in solution, it enables the analysis of heterogeneous populations of potentially unordered nanoparticles that are synthesized in solution, thereby providing a means to understand the structure and stability of defects at the nanoscale. PMID:26185247

  3. 3D structure of individual nanocrystals in solution by electron microscopy

    NASA Astrophysics Data System (ADS)

    Park, Jungwon; Elmlund, Hans; Ercius, Peter; Yuk, Jong Min; Limmer, David T.; Chen, Qian; Kim, Kwanpyo; Han, Sang Hoon; Weitz, David A.; Zettl, A.; Alivisatos, A. Paul

    2015-07-01

    Knowledge about the synthesis, growth mechanisms, and physical properties of colloidal nanoparticles has been limited by technical impediments. We introduce a method for determining three-dimensional (3D) structures of individual nanoparticles in solution. We combine a graphene liquid cell, high-resolution transmission electron microscopy, a direct electron detector, and an algorithm for single-particle 3D reconstruction originally developed for analysis of biological molecules. This method yielded two 3D structures of individual platinum nanocrystals at near-atomic resolution. Because our method derives the 3D structure from images of individual nanoparticles rotating freely in solution, it enables the analysis of heterogeneous populations of potentially unordered nanoparticles that are synthesized in solution, thereby providing a means to understand the structure and stability of defects at the nanoscale.

  4. Optical clearing based cellular-level 3D visualization of intact lymph node cortex

    PubMed Central

    Song, Eunjoo; Seo, Howon; Choe, Kibaek; Hwang, Yoonha; Ahn, Jinhyo; Ahn, Soyeon; Kim, Pilhan

    2015-01-01

    Lymph node (LN) is an important immune organ that controls adaptive immune responses against foreign pathogens and abnormal cells. To facilitate efficient immune function, LN has highly organized 3D cellular structures, vascular and lymphatic system. Unfortunately, conventional histological analysis relying on thin-sliced tissue has limitations in 3D cellular analysis due to structural disruption and tissue loss in the processes of fixation and tissue slicing. Optical sectioning confocal microscopy has been utilized to analyze 3D structure of intact LN tissue without physical tissue slicing. However, light scattering within biological tissues limits the imaging depth only to superficial portion of LN cortex. Recently, optical clearing techniques have shown enhancement of imaging depth in various biological tissues, but their efficacy for LN are remained to be investigated. In this work, we established optical clearing procedure for LN and achieved 3D volumetric visualization of the whole cortex of LN. More than 4 times improvement in imaging depth was confirmed by using LN obtained from H2B-GFP/actin-DsRed double reporter transgenic mouse. With adoptive transfer of GFP expressing B cells and DsRed expressing T cells and fluorescent vascular labeling by anti-CD31 and anti-LYVE-1 antibody conjugates, we successfully visualized major cellular-level structures such as T-cell zone, B-cell follicle and germinal center. Further, we visualized the GFP expressing metastatic melanoma cell colony, vasculature and lymphatic vessels in the LN cortex. PMID:26504662

  5. Rapid, High-Throughput Tracking of Bacterial Motility in 3D via Phase-Contrast Holographic Video Microscopy

    PubMed Central

    Cheong, Fook Chiong; Wong, Chui Ching; Gao, YunFeng; Nai, Mui Hoon; Cui, Yidan; Park, Sungsu; Kenney, Linda J.; Lim, Chwee Teck

    2015-01-01

    Tracking fast-swimming bacteria in three dimensions can be extremely challenging with current optical techniques and a microscopic approach that can rapidly acquire volumetric information is required. Here, we introduce phase-contrast holographic video microscopy as a solution for the simultaneous tracking of multiple fast moving cells in three dimensions. This technique uses interference patterns formed between the scattered and the incident field to infer the three-dimensional (3D) position and size of bacteria. Using this optical approach, motility dynamics of multiple bacteria in three dimensions, such as speed and turn angles, can be obtained within minutes. We demonstrated the feasibility of this method by effectively tracking multiple bacteria species, including Escherichia coli, Agrobacterium tumefaciens, and Pseudomonas aeruginosa. In addition, we combined our fast 3D imaging technique with a microfluidic device to present an example of a drug/chemical assay to study effects on bacterial motility. PMID:25762336

  6. Blind deconvolution of 3D fluorescence microscopy using depth-variant asymmetric PSF.

    PubMed

    Kim, Boyoung; Naemura, Takeshi

    2016-06-01

    The 3D wide-field fluorescence microscopy suffers from depth-variant asymmetric blur. The depth-variance and axial asymmetry are due to refractive index mismatch between the immersion and the specimen layer. The radial asymmetry is due to lens imperfections and local refractive index inhomogeneities in the specimen. To obtain the PSF that has these characteristics, there were PSF premeasurement trials. However, they are useless since imaging conditions such as camera position and refractive index of the specimen are changed between the premeasurement and actual imaging. In this article, we focus on removing unknown depth-variant asymmetric blur in such an optical system under the assumption of refractive index homogeneities in the specimen. We propose finding few parameters in the mathematical PSF model from observed images in which the PSF model has a depth-variant asymmetric shape. After generating an initial PSF from the analysis of intensities in the observed image, the parameters are estimated based on a maximum likelihood estimator. Using the estimated PSF, we implement an accelerated GEM algorithm for image deconvolution. Deconvolution result shows the superiority of our algorithm in terms of accuracy, which quantitatively evaluated by FWHM, relative contrast, standard deviation values of intensity peaks and FWHM. Microsc. Res. Tech. 79:480-494, 2016. © 2016 Wiley Periodicals, Inc. PMID:27062314

  7. Bridging microscopes: 3D correlative light and scanning electron microscopy of complex biological structures.

    PubMed

    Lucas, Miriam S; Günthert, Maja; Gasser, Philippe; Lucas, Falk; Wepf, Roger

    2012-01-01

    The rationale of correlative light and electron microscopy (CLEM) is to collect data on different information levels--ideally from an identical area on the same sample--with the aim of combining datasets at different levels of resolution to achieve a more holistic view of the hierarchical structural organization of cells and tissues. Modern three-dimensional (3D) imaging techniques in light and electron microscopy opened up new possibilities to expand morphological studies into the third dimension at the nanometer scale and over various volume dimensions. Here, we present two alternative approaches to correlate 3D light microscopy (LM) data with scanning electron microscopy (SEM) volume data. An adapted sample preparation method based on high-pressure freezing for structure preservation, followed by freeze-substitution for multimodal en-bloc imaging or serial-section imaging is described. The advantages and potential applications are exemplarily shown on various biological samples, such as cells, individual organisms, human tissue, as well as plant tissue. The two CLEM approaches presented here are per se not mutually exclusive, but have their distinct advantages. Confocal laser scanning microscopy (CLSM) and focused ion beam-SEM (FIB-SEM) is most suitable for targeted 3D correlation of small volumes, whereas serial-section LM and SEM imaging has its strength in large-area or -volume screening and correlation. The second method can be combined with immunocytochemical methods. Both methods, however, have the potential to extract statistically relevant data of structural details for systems biology.

  8. 3D high-density localization microscopy using hybrid astigmatic/ biplane imaging and sparse image reconstruction.

    PubMed

    Min, Junhong; Holden, Seamus J; Carlini, Lina; Unser, Michael; Manley, Suliana; Ye, Jong Chul

    2014-11-01

    Localization microscopy achieves nanoscale spatial resolution by iterative localization of sparsely activated molecules, which generally leads to a long acquisition time. By implementing advanced algorithms to treat overlapping point spread functions (PSFs), imaging of densely activated molecules can improve the limited temporal resolution, as has been well demonstrated in two-dimensional imaging. However, three-dimensional (3D) localization of high-density data remains challenging since PSFs are far more similar along the axial dimension than the lateral dimensions. Here, we present a new, high-density 3D imaging system and algorithm. The hybrid system is implemented by combining astigmatic and biplane imaging. The proposed 3D reconstruction algorithm is extended from our state-of-the art 2D high-density localization algorithm. Using mutual coherence analysis of model PSFs, we validated that the hybrid system is more suitable than astigmatic or biplane imaging alone for 3D localization of high-density data. The efficacy of the proposed method was confirmed via simulation and real data of microtubules. Furthermore, we also successfully demonstrated fluorescent-protein-based live cell 3D localization microscopy with a temporal resolution of just 3 seconds, capturing fast dynamics of the endoplasmic recticulum.

  9. Computer-aided microtomography with true 3-D display in electron microscopy.

    PubMed

    Nelson, A C

    1986-01-01

    A novel research system has been designed to permit three-dimensional (3-D) viewing of high resolution image data from transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The system consists of front-end primary data acquisition devices, such as TEM and SEM machines, which are equipped with computer-controlled specimen tilt stages. The output from these machines is in analogue form, where a video camera attached to the TEM provides the sequential analogue image output while the SEM direct video output is utilized. A 10 MHz digitizer transforms the video image to a digital array of 512 X 512 pixel units of 8 bits deep-stored in a frame buffer. Digital images from multiple projections are reconstructed into 3-D image boxes in a dedicated computer. Attached to the computer is a powerful true 3-D display device which has hardware for graphic manipulations including tilt and rotate on any axis and for probing the image with a 3-D cursor. Data editing and automatic contouring functions are used to enhance areas of interest, and specialized software is available for measurement of numbers, distances, areas, and volumes. With proper archiving of reconstructed image sequences, a dynamic 3-D presentation is possible. The microtomography system is highly versatile and can process image data on-line or from remote sites from which data records would typically be transported on computer tape, video tape, or floppy disk. PMID:3753610

  10. Fiber based optical tweezers for simultaneous in situ force exertion and measurements in a 3D polyacrylamide gel compartment.

    PubMed

    Ti, Chaoyang; Thomas, Gawain M; Ren, Yundong; Zhang, Rui; Wen, Qi; Liu, Yuxiang

    2015-07-01

    Optical tweezers play an important role in biological applications. However, it is difficult for traditional optical tweezers based on objective lenses to work in a three-dimensional (3D) solid far away from the substrate. In this work, we develop a fiber based optical trapping system, namely inclined dual fiber optical tweezers, that can simultaneously apply and measure forces both in water and in a 3D polyacrylamide gel matrix. In addition, we demonstrate in situ, non-invasive characterization of local mechanical properties of polyacrylamide gel by measurements on an embedded bead. The fiber optical tweezers measurements agree well with those of atomic force microscopy (AFM). The inclined dual fiber optical tweezers provide a promising and versatile tool for cell mechanics study in 3D environments.

  11. Fiber based optical tweezers for simultaneous in situ force exertion and measurements in a 3D polyacrylamide gel compartment

    PubMed Central

    Ti, Chaoyang; Thomas, Gawain M; Ren, Yundong; Zhang, Rui; Wen, Qi; Liu, Yuxiang

    2015-01-01

    Optical tweezers play an important role in biological applications. However, it is difficult for traditional optical tweezers based on objective lenses to work in a three-dimensional (3D) solid far away from the substrate. In this work, we develop a fiber based optical trapping system, namely inclined dual fiber optical tweezers, that can simultaneously apply and measure forces both in water and in a 3D polyacrylamide gel matrix. In addition, we demonstrate in situ, non-invasive characterization of local mechanical properties of polyacrylamide gel by measurements on an embedded bead. The fiber optical tweezers measurements agree well with those of atomic force microscopy (AFM). The inclined dual fiber optical tweezers provide a promising and versatile tool for cell mechanics study in 3D environments. PMID:26203364

  12. 3D imaging of the cleared intact murine colon with light sheet microscopy

    NASA Astrophysics Data System (ADS)

    Zufiria, B.; Bocancea, D. I.; Gómez-Gaviro, M. V.; Vaquero, J. J.; Desco, M.; Fresno, M.; Ripoll, J.; Arranz, A.

    2016-03-01

    We here show 3D light sheet microscopy images of fixed and cleared murine colon tissue in-toto, which offer relevant cellular information without the need for physically sectioning the tissue. We have applied the recently developed CUBIC protocol (Susaki et al. Cell 157:726, 2014) for colon tissues and have found that this clearing protocol enables imaging all the way to the central part of the lumen with cellular resolution, thus opening new ways for 3D imaging of colon samples.

  13. Generalized recovery algorithm for 3D super-resolution microscopy using rotating point spread functions

    NASA Astrophysics Data System (ADS)

    Shuang, Bo; Wang, Wenxiao; Shen, Hao; Tauzin, Lawrence J.; Flatebo, Charlotte; Chen, Jianbo; Moringo, Nicholas A.; Bishop, Logan D. C.; Kelly, Kevin F.; Landes, Christy F.

    2016-08-01

    Super-resolution microscopy with phase masks is a promising technique for 3D imaging and tracking. Due to the complexity of the resultant point spread functions, generalized recovery algorithms are still missing. We introduce a 3D super-resolution recovery algorithm that works for a variety of phase masks generating 3D point spread functions. A fast deconvolution process generates initial guesses, which are further refined by least squares fitting. Overfitting is suppressed using a machine learning determined threshold. Preliminary results on experimental data show that our algorithm can be used to super-localize 3D adsorption events within a porous polymer film and is useful for evaluating potential phase masks. Finally, we demonstrate that parallel computation on graphics processing units can reduce the processing time required for 3D recovery. Simulations reveal that, through desktop parallelization, the ultimate limit of real-time processing is possible. Our program is the first open source recovery program for generalized 3D recovery using rotating point spread functions.

  14. Generalized recovery algorithm for 3D super-resolution microscopy using rotating point spread functions

    PubMed Central

    Shuang, Bo; Wang, Wenxiao; Shen, Hao; Tauzin, Lawrence J.; Flatebo, Charlotte; Chen, Jianbo; Moringo, Nicholas A.; Bishop, Logan D. C.; Kelly, Kevin F.; Landes, Christy F.

    2016-01-01

    Super-resolution microscopy with phase masks is a promising technique for 3D imaging and tracking. Due to the complexity of the resultant point spread functions, generalized recovery algorithms are still missing. We introduce a 3D super-resolution recovery algorithm that works for a variety of phase masks generating 3D point spread functions. A fast deconvolution process generates initial guesses, which are further refined by least squares fitting. Overfitting is suppressed using a machine learning determined threshold. Preliminary results on experimental data show that our algorithm can be used to super-localize 3D adsorption events within a porous polymer film and is useful for evaluating potential phase masks. Finally, we demonstrate that parallel computation on graphics processing units can reduce the processing time required for 3D recovery. Simulations reveal that, through desktop parallelization, the ultimate limit of real-time processing is possible. Our program is the first open source recovery program for generalized 3D recovery using rotating point spread functions. PMID:27488312

  15. Generalized recovery algorithm for 3D super-resolution microscopy using rotating point spread functions.

    PubMed

    Shuang, Bo; Wang, Wenxiao; Shen, Hao; Tauzin, Lawrence J; Flatebo, Charlotte; Chen, Jianbo; Moringo, Nicholas A; Bishop, Logan D C; Kelly, Kevin F; Landes, Christy F

    2016-01-01

    Super-resolution microscopy with phase masks is a promising technique for 3D imaging and tracking. Due to the complexity of the resultant point spread functions, generalized recovery algorithms are still missing. We introduce a 3D super-resolution recovery algorithm that works for a variety of phase masks generating 3D point spread functions. A fast deconvolution process generates initial guesses, which are further refined by least squares fitting. Overfitting is suppressed using a machine learning determined threshold. Preliminary results on experimental data show that our algorithm can be used to super-localize 3D adsorption events within a porous polymer film and is useful for evaluating potential phase masks. Finally, we demonstrate that parallel computation on graphics processing units can reduce the processing time required for 3D recovery. Simulations reveal that, through desktop parallelization, the ultimate limit of real-time processing is possible. Our program is the first open source recovery program for generalized 3D recovery using rotating point spread functions.

  16. Generalized recovery algorithm for 3D super-resolution microscopy using rotating point spread functions.

    PubMed

    Shuang, Bo; Wang, Wenxiao; Shen, Hao; Tauzin, Lawrence J; Flatebo, Charlotte; Chen, Jianbo; Moringo, Nicholas A; Bishop, Logan D C; Kelly, Kevin F; Landes, Christy F

    2016-01-01

    Super-resolution microscopy with phase masks is a promising technique for 3D imaging and tracking. Due to the complexity of the resultant point spread functions, generalized recovery algorithms are still missing. We introduce a 3D super-resolution recovery algorithm that works for a variety of phase masks generating 3D point spread functions. A fast deconvolution process generates initial guesses, which are further refined by least squares fitting. Overfitting is suppressed using a machine learning determined threshold. Preliminary results on experimental data show that our algorithm can be used to super-localize 3D adsorption events within a porous polymer film and is useful for evaluating potential phase masks. Finally, we demonstrate that parallel computation on graphics processing units can reduce the processing time required for 3D recovery. Simulations reveal that, through desktop parallelization, the ultimate limit of real-time processing is possible. Our program is the first open source recovery program for generalized 3D recovery using rotating point spread functions. PMID:27488312

  17. Video lensfree microscopy of 2D and 3D culture of cells

    NASA Astrophysics Data System (ADS)

    Allier, C. P.; Vinjimore Kesavan, S.; Coutard, J.-G.; Cioni, O.; Momey, F.; Navarro, F.; Menneteau, M.; Chalmond, B.; Obeid, P.; Haguet, V.; David-Watine, B.; Dubrulle, N.; Shorte, S.; van der Sanden, B.; Di Natale, C.; Hamard, L.; Wion, D.; Dolega, M. E.; Picollet-D'hahan, N.; Gidrol, X.; Dinten, J.-M.

    2014-03-01

    Innovative imaging methods are continuously developed to investigate the function of biological systems at the microscopic scale. As an alternative to advanced cell microscopy techniques, we are developing lensfree video microscopy that opens new ranges of capabilities, in particular at the mesoscopic level. Lensfree video microscopy allows the observation of a cell culture in an incubator over a very large field of view (24 mm2) for extended periods of time. As a result, a large set of comprehensive data can be gathered with strong statistics, both in space and time. Video lensfree microscopy can capture images of cells cultured in various physical environments. We emphasize on two different case studies: the quantitative analysis of the spontaneous network formation of HUVEC endothelial cells, and by coupling lensfree microscopy with 3D cell culture in the study of epithelial tissue morphogenesis. In summary, we demonstrate that lensfree video microscopy is a powerful tool to conduct cell assays in 2D and 3D culture experiments. The applications are in the realms of fundamental biology, tissue regeneration, drug development and toxicology studies.

  18. Investigation on 3D morphological changes of in vitro cells through digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Memmolo, Pasquale; Miccio, Lisa; Merola, Francesco; Netti, Paolo A.; Coppola, Giuseppe; Ferraro, Pietro

    2013-04-01

    We report the investigation of the identification and measurement of region of interest (ROI) in quantitative phase-contrast maps (QPMs) of biological cells by digital holographic microscopy (DHM), with the aim to analyze the 3D positions and 3D morphology together. We consider as test case for our tool the in vitro bull sperm head morphometry analysis. Extraction and measurement of various morphological parameters are performed by using two methods: the anisotropic diffusion filter, that is based on the Gaussian diffusivity function which allows more accuracy of the edge position, and the simple thresholding filter. In particular we consider the calculation of area, ellipticity, perimeter, major axis, minor axis and shape factor as a morphological parameter, instead, for the estimation of 3D position, we compute the centroid, the weighted centroid and the maximum phase values. A statistical analysis on a data set composed by N = 14 holograms relative to bovine spermatozoa and its reference holograms is reported.

  19. Optical 3D-coordinate measuring system using structured light

    NASA Astrophysics Data System (ADS)

    Schreiber, Wolfgang; Notni, Gunther; Kuehmstedt, Peter; Gerber, Joerg; Kowarschik, Richard M.

    1996-09-01

    The paper is aimed at the description of an optical shape measuring technique based on a consistent principle using fringe projection technique. We demonstrate a real 3D- coordinate measuring system where the sale of coordinates is given only by the illumination-structures. This method has the advantages that the aberration of the observing system and the depth-dependent imaging scale have no influence on the measuring accuracy and, moreover, the measurements are independent of the position of the camera with respect to the object under test. Furthermore, it is shown that the influence of specular effects of the surface on the measuring result can be eliminated. Moreover, we developed a very simple algorithm to calibrate the measuring system. The measuring examples show that a measuring accuracy of 10-4 (i.e. 10 micrometers ) within an object volume of 100 X 100 X 70 mm3 is achievable. Furthermore, it is demonstrated that the set of coordinate values can be processed in CNC- and CAD-systems.

  20. Cordless hand-held optical 3D sensor

    NASA Astrophysics Data System (ADS)

    Munkelt, Christoph; Bräuer-Burchardt, Christian; Kühmstedt, Peter; Schmidt, Ingo; Notni, Gunther

    2007-07-01

    A new mobile optical 3D measurement system using phase correlation based fringe projection technique will be presented. The sensor consist of a digital projection unit and two cameras in a stereo arrangement, whereby both are battery powered. The data transfer to a base station will be done via WLAN. This gives the possibility to use the system in complicate, remote measurement situations, which are typical in archaeology and architecture. In the measurement procedure the sensor will be hand-held by the user, illuminating the object with a sequence of less than 10 fringe patterns, within a time below 200 ms. This short sequence duration was achieved by a new approach, which combines the epipolar constraint with robust phase correlation utilizing a pre-calibrated sensor head, containing two cameras and a digital fringe projector. Furthermore, the system can be utilized to acquire the all around shape of objects by using the phasogrammetric approach with virtual land marks introduced by the authors 1, 2. This way no matching procedures or markers are necessary for the registration of multiple views, which makes the system very flexible in accomplishing different measurement tasks. The realized measurement field is approx. 100 mm up to 400 mm in diameter. The mobile character makes the measurement system useful for a wide range of applications in arts, architecture, archaeology and criminology, which will be shown in the paper.

  1. 3D print customized sample holders for live light sheet microscopy.

    PubMed

    Jeandupeux, Emeric; Lobjois, Valérie; Ducommun, Bernard

    2015-08-01

    A major hurdle to the widespread application of light sheet microscopy is the lack of versatile and non-intrusive sample holders that are adaptable to a variety of biological samples for live imaging. To overcome this limitation, we present herein the application of 3D printing to the fabrication of a fully customizable casting kit. 3D printing enables facile preparation of hydrogel sample holders adaptable to any shape and number of specimen. As an example, we present the use of this device to produce a four-sample holder adapted to parallel live monitoring of multicellular tumor spheroid growth. To share our solution with the light sheet microscopy community, all files necessary to produce or customize sample holders are freely available online.

  2. 3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy.

    PubMed

    Rennie, Monique Y; Gahan, Curran G; López, Claudia S; Thornburg, Kent L; Rugonyi, Sandra

    2014-08-01

    Early embryonic heart development is a period of dynamic growth and remodeling, with rapid changes occurring at the tissue, cell, and subcellular levels. A detailed understanding of the events that establish the components of the heart wall has been hampered by a lack of methodologies for three-dimensional (3D), high-resolution imaging. Focused ion beam scanning electron microscopy (FIB-SEM) is a novel technology for imaging 3D tissue volumes at the subcellular level. FIB-SEM alternates between imaging the block face with a scanning electron beam and milling away thin sections of tissue with a FIB, allowing for collection and analysis of 3D data. FIB-SEM was used to image the three layers of the day 4 chicken embryo heart: myocardium, cardiac jelly, and endocardium. Individual images obtained with FIB-SEM were comparable in quality and resolution to those obtained with transmission electron microscopy. Up to 1,100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800-1,500 μm3 in size. Segmentation of organelles revealed their organization and distinct volume fractions between cardiac wall layers. We conclude that FIB-SEM is a powerful modality for 3D subcellular imaging of the embryonic heart wall.

  3. 3D single molecule tracking in thick cellular specimens using multifocal plane microscopy

    NASA Astrophysics Data System (ADS)

    Ram, Sripad; Ward, E. Sally; Ober, Raimund J.

    2011-03-01

    One of the major challenges in single molecule microscopy concerns 3D tracking of single molecules in cellular specimens. This has been a major impediment to study many fundamental cellular processes, such as protein transport across thick cellular specimens (e.g. a cell-monolayer). Here we show that multifocal plane microscopy (MUM), an imaging modality developed by our group, provides the much needed solution to this longstanding problem. While MUM was previously used for 3D single molecule tracking at shallow depths (~ 1 micron) in live-cells, the question arises if MUM can also live up to the significant challenge of tracking single molecules in thick samples. Here by substantially expanding the capabilities of MUM, we demonstrate 3D tracking of quantum-dot labeled molecules in a ~ 10 micron thick cell monolayer. In this way we have reconstructed the complete 3D intracellular trafficking itinerary of single molecules at high spatial and temporal precision in a thick cell-sample. Funding support: NIH and the National MS Society.

  4. Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell

    PubMed Central

    Bohórquez, Diego; Haque, Fariha; Medicetty, Satish; Liddle, Rodger A.

    2015-01-01

    Delineation of a cell’s ultrastructure is important for understanding its function. This can be a daunting project for rare cell types diffused throughout tissues made of diverse cell types, such as enteroendocrine cells of the intestinal epithelium. These gastrointestinal sensors of food and bacteria have been difficult to study because they are dispersed among other epithelial cells at a ratio of 1:1,000. Recently, transgenic reporter mice have been generated to identify enteroendocrine cells by means of fluorescence. One of those is the peptide YY-GFP mouse. Using this mouse, we developed a method to correlate confocal and serial block-face scanning electron microscopy. We named the method cocem3D and applied it to identify a specific enteroendocrine cell in tissue and unveil the cell’s ultrastructure in 3D. The resolution of cocem3D is sufficient to identify organelles as small as secretory vesicles and to distinguish cell membranes for volume rendering. Cocem3D can be easily adapted to study the 3D ultrastructure of other specific cell types in their native tissue. PMID:26273796

  5. Virtual rough samples to test 3D nanometer-scale scanning electron microscopy stereo photogrammetry

    NASA Astrophysics Data System (ADS)

    Villarrubia, J. S.; Tondare, V. N.; Vladár, A. E.

    2016-03-01

    The combination of scanning electron microscopy for high spatial resolution, images from multiple angles to provide 3D information, and commercially available stereo photogrammetry software for 3D reconstruction offers promise for nanometer-scale dimensional metrology in 3D. A method is described to test 3D photogrammetry software by the use of virtual samples—mathematical samples from which simulated images are made for use as inputs to the software under test. The virtual sample is constructed by wrapping a rough skin with any desired power spectral density around a smooth near-trapezoidal line with rounded top corners. Reconstruction is performed with images simulated from different angular viewpoints. The software's reconstructed 3D model is then compared to the known geometry of the virtual sample. Three commercial photogrammetry software packages were tested. Two of them produced results for line height and width that were within close to 1 nm of the correct values. All of the packages exhibited some difficulty in reconstructing details of the surface roughness.

  6. Trapped particle optical microscopy

    NASA Astrophysics Data System (ADS)

    Malmqvist, Lars; Hertz, Hans M.

    1992-11-01

    A scanned probe optical microscope allowing nondestructive studies of a wide range of objects and surface is described. The microscope utilizes a noninvasive optical trap to position a microscopic probe light source in immediate proximity to the studied object. We demonstrate the method experimentally and show theoretically its potential for optical imaging with subdiffraction limited resolution of, e.g., biological objects.

  7. In vivo multiphoton microscopy associated to 3D image processing for human skin characterization

    NASA Astrophysics Data System (ADS)

    Baldeweck, T.; Tancrède, E.; Dokladal, P.; Koudoro, S.; Morard, V.; Meyer, F.; Decencière, E.; Pena, A.-M.

    2012-03-01

    Multiphoton microscopy has emerged in the past decade as a promising non-invasive skin imaging technique. The aim of this study was to assess whether multiphoton microscopy coupled to specific 3D image processing tools could provide new insights into the organization of different skin components and their age-related changes. For that purpose, we performed a clinical trial on 15 young and 15 aged human female volunteers on the ventral and dorsal side of the forearm using the DermaInspectR medical imaging device. We visualized the skin by taking advantage of intrinsic multiphoton signals from cells, elastic and collagen fibers. We also developed 3D image processing algorithms adapted to in vivo multiphoton images of human skin in order to extract quantitative parameters in each layer of the skin (epidermis and superficial dermis). The results show that in vivo multiphoton microscopy is able to evidence several skin alterations due to skin aging: morphological changes in the epidermis and modifications in the quantity and organization of the collagen and elastic fibers network. In conclusion, the association of multiphoton microscopy with specific image processing allows the three-dimensional organization of skin components to be visualized and quantified thus providing a powerful tool for cosmetic and dermatological investigations.

  8. 3D imaging and characterization of microlenses and microlens arrays using nonlinear microscopy

    NASA Astrophysics Data System (ADS)

    Krmpot, Aleksandar J.; Tserevelakis, George J.; Murić, Branka D.; Filippidis, George; Pantelić, Dejan V.

    2013-05-01

    In this work, nonlinear laser scanning microscopy was employed for the characterization and three-dimensional (3D) imaging of microlenses and microlens arrays. Third-harmonic generation and two-photon excitation fluorescence (TPEF) signals were recorded and the obtained data were further processed in order to generate 3D reconstructions of the examined samples. Femtosecond laser pulses (1028 nm) were utilized for excitation. Microlenses were manufactured on Tot'hema and eosin sensitized gelatin layers using a green (532 nm) continuous wave laser beam using the direct laser writing method. The profiles of the microlens surface were obtained from the radial cross-sections, using a triple-Gaussian fit. The analytical shapes of the profiles were also used for ray tracing. Furthermore, the volumes of the microlenses were determined with high precision. The TPEF signal arising from the volume of the material was recorded and the respective 3D spatial fluorescence distribution of the samples was mapped. Nonlinear microscopy modalities have been shown to be a powerful diagnostic tool for microlens characterization as they enable in-depth investigations of the structural properties of the samples, in a nondestructive manner.

  9. Air-structured optical fiber drawn from a 3D-printed preform.

    PubMed

    Cook, Kevin; Canning, John; Leon-Saval, Sergio; Reid, Zane; Hossain, Md Arafat; Comatti, Jade-Edouard; Luo, Yanhua; Peng, Gang-Ding

    2015-09-01

    A structured optical fiber is drawn from a 3D-printed structured preform. Preforms containing a single ring of holes around the core are fabricated using filament made from a modified butadiene polymer. More broadly, 3D printers capable of processing soft glasses, silica, and other materials are likely to come on line in the not-so-distant future. 3D printing of optical preforms signals a new milestone in optical fiber manufacture.

  10. Cell cycle phase classification in 3D in vivo microscopy of Drosophila embryogenesis

    PubMed Central

    2011-01-01

    Background Cell divisions play critical roles in disease and development. The analysis of cell division phenotypes in high content image-based screening and time-lapse microscopy relies on automated nuclear segmentation and classification of cell cycle phases. Automated identification of the cell cycle phase helps biologists quantify the effect of genetic perturbations and drug treatments. Most existing studies have dealt with 2D images of cultured cells. Few, if any, studies have addressed the problem of cell cycle classification in 3D image stacks of intact tissues. Results We developed a workflow for the automated cell cycle phase classification in 3D time-series image datasets of live Drosophila embryos expressing the chromatin marker histone-GFP. Upon image acquisition by laser scanning confocal microscopy and 3D nuclear segmentation, we extracted 3D intensity, shape and texture features from interphase nuclei and mitotic chromosomes. We trained different classifiers, including support vector machines (SVM) and neural networks, to distinguish between 5 cell cycles phases (Interphase and 4 mitotic phases) and achieved over 90% accuracy. As the different phases occur at different frequencies (58% of samples correspond to interphase), we devised a strategy to improve the identification of classes with low representation. To investigate which features are required for accurate classification, we performed feature reduction and selection. We were able to reduce the feature set from 42 to 9 without affecting classifier performance. We observed a dramatic decrease of classification performance when the training and testing samples were derived from two different developmental stages, the nuclear divisions of the syncytial blastoderm and the cell divisions during gastrulation. Combining samples from both developmental stages produced a more robust and accurate classifier. Conclusions Our study demonstrates that automated cell cycle phase classification, besides 2D

  11. 3D structure tensor analysis of light microscopy data for validating diffusion MRI.

    PubMed

    Khan, Ahmad Raza; Cornea, Anda; Leigland, Lindsey A; Kohama, Steven G; Jespersen, Sune Nørhøj; Kroenke, Christopher D

    2015-05-01

    Diffusion magnetic resonance imaging (d-MRI) is a powerful non-invasive and non-destructive technique for characterizing brain tissue on the microscopic scale. However, the lack of validation of d-MRI by independent experimental means poses an obstacle to accurate interpretation of data acquired using this method. Recently, structure tensor analysis has been applied to light microscopy images, and this technique holds promise to be a powerful validation strategy for d-MRI. Advantages of this approach include its similarity to d-MRI in terms of averaging the effects of a large number of cellular structures, and its simplicity, which enables it to be implemented in a high-throughput manner. However, a drawback of previous implementations of this technique arises from it being restricted to 2D. As a result, structure tensor analyses have been limited to tissue sectioned in a direction orthogonal to the direction of interest. Here we describe the analytical framework for extending structure tensor analysis to 3D, and utilize the results to analyze serial image "stacks" acquired with confocal microscopy of rhesus macaque hippocampal tissue. Implementation of 3D structure tensor procedures requires removal of sources of anisotropy introduced in tissue preparation and confocal imaging. This is accomplished with image processing steps to mitigate the effects of anisotropic tissue shrinkage, and the effects of anisotropy in the point spread function (PSF). In order to address the latter confound, we describe procedures for measuring the dependence of PSF anisotropy on distance from the microscope objective within tissue. Prior to microscopy, ex vivo d-MRI measurements performed on the hippocampal tissue revealed three regions of tissue with mutually orthogonal directions of least restricted diffusion that correspond to CA1, alveus and inferior longitudinal fasciculus. We demonstrate the ability of 3D structure tensor analysis to identify structure tensor orientations that

  12. 3D structure tensor analysis of light microscopy data for validating diffusion MRI

    PubMed Central

    Khan, Ahmad Raza; Cornea, Anda; Leigland, Lindsey A.; Kohama, Steven G.; Jespersen, Sune Nørhøj; Kroenke, Christopher D.

    2015-01-01

    Diffusion magnetic resonance imaging (d-MRI) is a powerful non-invasive and non-destructive technique for characterizing brain tissue on the microscopic scale. However, the lack of validation of d-MRI by independent experimental means poses an obstacle to accurate interpretation of data acquired using this method. Recently, structure tensor analysis has been applied to light microscopy images, and this technique holds promise to be a powerful validation strategy for d-MRI. Advantages of this approach include its similarity to d-MRI in terms of averaging the effects of a large number of cellular structures, and its simplicity, which enables it to be implemented in a high-throughput manner. However, a drawback of previous implementations of this technique arises from it being restricted to 2D. As a result, structure tensor analyses have been limited to tissue sectioned in a direction orthogonal to the direction of interest. Here we describe the analytical framework for extending structure tensor analysis to 3D, and utilize the results to analyze serial image “stacks” acquired with confocal microscopy of rhesus macaque hippocampal tissue. Implementation of 3D structure tensor procedures requires removal of sources of anisotropy introduced in tissue preparation and confocal imaging. This is accomplished with image processing steps to mitigate the effects of anisotropic tissue shrinkage, and the effects of anisotropy in the point spread function (PSF). In order to address the latter confound, we describe procedures for measuring the dependence of PSF anisotropy on distance from the microscope objective within tissue. Prior to microscopy, ex vivo d-MRI measurements performed on the hippocampal tissue revealed three regions of tissue with mutually orthogonal directions of least restricted diffusion that correspond to CA1, alveus and inferior longitudinal fasciculus. We demonstrate the ability of 3D structure tensor analysis to identify structure tensor orientations

  13. Application issues when using optical 3D systems in place of CMMs

    NASA Astrophysics Data System (ADS)

    Harding, Kevin G.

    2002-02-01

    A primary application of optical 3D measurement systems has been in the replacement of mechanical coordinate measurement machines (CMMs). The advantage of optical 3D systems is typically greater speed and flexibility of operation over even the best CMMs. However, the two technologies are not necessarily one to one replacements, requiring new methods of use and in general proof of performance. This paper will present specific data that highlights the differences between CMMs and optical 3D systems and suggests a method to properly achieve CMM compatible results with an optical 3D system and the problems seen in this study.

  14. Subcellular Microanatomy by 3D Deconvolution Brightfield Microscopy: Method and Analysis Using Human Chromatin in the Interphase Nucleus

    PubMed Central

    Tadrous, Paul Joseph

    2012-01-01

    Anatomy has advanced using 3-dimensional (3D) studies at macroscopic (e.g., dissection, injection moulding of vessels, radiology) and microscopic (e.g., serial section reconstruction with light and electron microscopy) levels. This paper presents the first results in human cells of a new method of subcellular 3D brightfield microscopy. Unlike traditional 3D deconvolution and confocal techniques, this method is suitable for general application to brightfield microscopy. Unlike brightfield serial sectioning it has subcellular resolution. Results are presented of the 3D structure of chromatin in the interphase nucleus of two human cell types, hepatocyte and plasma cell. I show how the freedom to examine these structures in 3D allows greater morphological discrimination between and within cell types and the 3D structural basis for the classical “clock-face” motif of the plasma cell nucleus is revealed. Potential for further applications discussed. PMID:22567315

  15. Scipion: A software framework toward integration, reproducibility and validation in 3D electron microscopy.

    PubMed

    de la Rosa-Trevín, J M; Quintana, A; Del Cano, L; Zaldívar, A; Foche, I; Gutiérrez, J; Gómez-Blanco, J; Burguet-Castell, J; Cuenca-Alba, J; Abrishami, V; Vargas, J; Otón, J; Sharov, G; Vilas, J L; Navas, J; Conesa, P; Kazemi, M; Marabini, R; Sorzano, C O S; Carazo, J M

    2016-07-01

    In the past few years, 3D electron microscopy (3DEM) has undergone a revolution in instrumentation and methodology. One of the central players in this wide-reaching change is the continuous development of image processing software. Here we present Scipion, a software framework for integrating several 3DEM software packages through a workflow-based approach. Scipion allows the execution of reusable, standardized, traceable and reproducible image-processing protocols. These protocols incorporate tools from different programs while providing full interoperability among them. Scipion is an open-source project that can be downloaded from http://scipion.cnb.csic.es. PMID:27108186

  16. Scipion: A software framework toward integration, reproducibility and validation in 3D electron microscopy.

    PubMed

    de la Rosa-Trevín, J M; Quintana, A; Del Cano, L; Zaldívar, A; Foche, I; Gutiérrez, J; Gómez-Blanco, J; Burguet-Castell, J; Cuenca-Alba, J; Abrishami, V; Vargas, J; Otón, J; Sharov, G; Vilas, J L; Navas, J; Conesa, P; Kazemi, M; Marabini, R; Sorzano, C O S; Carazo, J M

    2016-07-01

    In the past few years, 3D electron microscopy (3DEM) has undergone a revolution in instrumentation and methodology. One of the central players in this wide-reaching change is the continuous development of image processing software. Here we present Scipion, a software framework for integrating several 3DEM software packages through a workflow-based approach. Scipion allows the execution of reusable, standardized, traceable and reproducible image-processing protocols. These protocols incorporate tools from different programs while providing full interoperability among them. Scipion is an open-source project that can be downloaded from http://scipion.cnb.csic.es.

  17. Design of 3D isotropic metamaterial device using smart transformation optics.

    PubMed

    Shin, Dongheok; Kim, Junhyun; Yoo, Do-Sik; Kim, Kyoungsik

    2015-08-24

    We report here a design method for a 3 dimensional (3D) isotropic transformation optical device using smart transformation optics. Inspired by solid mechanics, smart transformation optics regards a transformation optical medium as an elastic solid and deformations as coordinate transformations. Further developing from our previous work on 2D smart transformation optics, we introduce a method of 3D smart transformation optics to design 3D transformation optical devices by maintaining isotropic materials properties for all types of polarizations imposing free or nearly free boundary conditions. Due to the material isotropy, it is possible to fabricate such devices with structural metamaterials made purely of common dielectric materials. In conclusion, the practical importance of the method reported here lies in the fact that it enables us to fabricate, without difficulty, arbitrarily shaped 3D devices with existing 3D printing technology.

  18. Design of 3D isotropic metamaterial device using smart transformation optics.

    PubMed

    Shin, Dongheok; Kim, Junhyun; Yoo, Do-Sik; Kim, Kyoungsik

    2015-08-24

    We report here a design method for a 3 dimensional (3D) isotropic transformation optical device using smart transformation optics. Inspired by solid mechanics, smart transformation optics regards a transformation optical medium as an elastic solid and deformations as coordinate transformations. Further developing from our previous work on 2D smart transformation optics, we introduce a method of 3D smart transformation optics to design 3D transformation optical devices by maintaining isotropic materials properties for all types of polarizations imposing free or nearly free boundary conditions. Due to the material isotropy, it is possible to fabricate such devices with structural metamaterials made purely of common dielectric materials. In conclusion, the practical importance of the method reported here lies in the fact that it enables us to fabricate, without difficulty, arbitrarily shaped 3D devices with existing 3D printing technology. PMID:26368165

  19. 3D Quantitative Confocal Laser Microscopy of Ilmenite Volume Distribution in Alpe Arami Olivine

    NASA Astrophysics Data System (ADS)

    Bozhilov, K. N.

    2001-12-01

    The deep origin of the Alpe Arami garnet lherzolite massif in the Swiss Alps proposed by Dobrzhinetskaya et al. (Science, 1996) has been a focus of heated debate. One of the lines of evidence supporting an exhumation from more than 200 km depth includes the abundance, distribution, and orientation of magnesian ilmenite rods in the oldest generation of olivine. This argument has been disputed in terms of the abundance of ilmenite and consequently the maximum TiO2 content in the discussed olivine. In order to address this issue, we have directly measured the volume fraction of ilmenite of the oldest generation of olivine by applying confocal laser scanning microscopy (CLSM). CLSM is a method which allows for three-dimensional imaging and quantitative volume determination by optical sectioning of the objects. The images for 3D reconstruction and measurements were acquired from petrographic thin sections in reflected laser light with 488 nm wavelength. Measurements of more than 80 olivine grains in six thin sections of our material yielded an average volume fraction of 0.31% ilmenite in the oldest generation of olivine from Alpe Arami. This translates into 0.23 wt.% TiO2 in olivine with error in determination of ±0.097 wt.%, a value significantly different from that of 0.02 to 0.03 wt.% TiO2 determined by Hacker et al. (Science, 1997) by a broad-beam microanalysis technique. During the complex geological history of the Alpe Arami massif, several events of metamorphism are recorded which all could have caused increased mobility of the mineral components. Evidence for loss of TiO2 from olivine is the tendency for high densities of ilmenite to be restricted to cores of old grains, the complete absence of ilmenite inclusions from the younger, recrystallized, generation of olivine, and reduction in ilmenite size and abundance in more serpentinized specimens. These observations suggest that only olivine grains with the highest concentrations of ilmenite are close to the

  20. The use of Interferometric Microscopy to assess 3D modifications of deteriorated medieval glass.

    NASA Astrophysics Data System (ADS)

    Gentaz, L.; Lombardo, T.; Chabas, A.

    2012-04-01

    Due to low durability, Northern European medieval glass undergoes the action of the atmospheric environment leading in some cases to a state of dramatic deterioration. Modification features varies from a simple loss of transparency to a severe material loss. In order to understand the underlying mechanisms and preserve this heritage, fundamental research is necessary too. In this optic, field exposure of analogues and original stained glass was carried out to study the early stages of the glass weathering. Model glass and original stained glass (after removal of deterioration products) were exposed in real conditions in an urban site (Paris) for 48 months. A regular withdrawal of samples allowed a follow-up of short-term glass evolution. Morphological modifications of the exposed samples were investigated through conventional and non destructive microscopy, using respectively a Scanning Electron Microscope (SEM) and an Interferometric Microscope (IM). This latter allows a 3D quantification of the object with no sample preparation. For all glasses, both surface recession and build-up of deposit were observed as a consequence of a leaching process (interdiffusion of protons and glass cations). The build-up of a deposit comes from the reaction between the extracted glass cations and atmospheric gases. Instead, surface recession is due mainly to the formation of brittle layer of altered glass at the sub-surface, where a fracture network can appear, leading to the scaling of parts of this modified glass. Finally, dissolution of the glass takes place, inducing the formation of pits and craters. The arithmetic roughness (Ra) was used as an indicator of weathering increase, in order to evaluate the deterioration state. For instance, the Ra grew from few tens of nm for pristine glass to thousands of nm for scaled areas. This technique also allowed a precise quantification of dimensions (height, depth and width) of deposits and pits, and the estimation of their overall

  1. Application of an optical 3D sensor for automated disassembling

    NASA Astrophysics Data System (ADS)

    Knackfuss, Peter; Schmidt, Achim

    1996-08-01

    The application of an active vision 3D sensor is described for the development and control of an autonomous intelligent robot cell for the disassembling of end-of-life-vehicle components. The research and development work was done concurrently by three European development teams at different locations. During this phase, the virtual environment was distributed on the local development platforms of these teams. Intermediate development results and 3D sensor data were exchanged through network communication to be mutually tested and verified. The physical environment of the disassembling cell demonstrator and its sensor systems is currently being integrated at the BIBA institute.

  2. Quantitative 3D molecular cutaneous absorption in human skin using label free nonlinear microscopy.

    PubMed

    Chen, Xueqin; Grégoire, Sébastien; Formanek, Florian; Galey, Jean-Baptiste; Rigneault, Hervé

    2015-02-28

    Understanding the penetration mechanisms of drugs into human skin is a key issue in pharmaceutical and cosmetics research. To date, the techniques available for percutaneous penetration of compounds fail to provide a quantitative 3D map of molecular concentration distribution in complex tissues as the detected microscopy images are an intricate combination of concentration distribution and laser beam attenuation upon deep penetration. Here we introduce and validate a novel framework for imaging and reconstructing molecular concentration within the depth of artificial and human skin samples. Our approach combines the use of deuterated molecular compounds together with coherent anti-Stokes Raman scattering spectroscopy and microscopy that permits targeted molecules to be unambiguously discriminated within skin layers. We demonstrate both intercellular and transcellular pathways for different active compounds, together with in-depth concentration profiles reflecting the detailed skin barrier architecture. This method provides an enabling platform for establishing functional activity of topically applied products. PMID:25550155

  3. Cellulose Nanocrystals as Chiral Inducers: Enantioselective Catalysis and Transmission Electron Microscopy 3D Characterization.

    PubMed

    Kaushik, Madhu; Basu, Kaustuv; Benoit, Charles; Cirtiu, Ciprian M; Vali, Hojatollah; Moores, Audrey

    2015-05-20

    Cellulose nanocrystals (CNCs), derived from cellulose, provide us with an opportunity to devise more sustainable solutions to current technological challenges. Enantioselective catalysis, especially heterogeneous, is the preferred method for the synthesis of pure chiral molecules in the fine chemical industries. Cellulose has been long sought as a chiral inducer in enantioselective catalysis. We report herein an unprecedentedly high enantiomeric excess (ee) for Pd patches deposited onto CNCs used as catalysts for the hydrogenation of prochiral ketones in water at room temperature and 4 bar H2. Our system, where CNCs acted as support and sole chiral source, achieved an ee of 65% with 100% conversions. Cryo-electron microscopy, high-resolution transmission electron microscopy, and tomography were used for the first time to study the 3D structure of a metal functionalized CNC hybrid. It established the presence of sub-nanometer-thick Pd patches at the surface of CNCs and provided insight into the chiral induction mechanism.

  4. Quantitative optical phase microscopy.

    PubMed

    Barty, A; Nugent, K A; Paganin, D; Roberts, A

    1998-06-01

    We present a new method for the extraction of quantitative phase data from microscopic phase samples by use of partially coherent illumination and an ordinary transmission microscope. The technique produces quantitative images of the phase profile of the sample without phase unwrapping. The technique is able to recover phase even in the presence of amplitude modulation, making it significantly more powerful than existing methods of phase microscopy. We demonstrate the technique by providing quantitatively correct phase images of well-characterized test samples and show that the results obtained for more-complex samples correlate with structures observed with Nomarski differential interference contrast techniques.

  5. X-ray microscopy for in situ characterization of 3D nanostructural evolution in the laboratory

    NASA Astrophysics Data System (ADS)

    Hornberger, Benjamin; Bale, Hrishikesh; Merkle, Arno; Feser, Michael; Harris, William; Etchin, Sergey; Leibowitz, Marty; Qiu, Wei; Tkachuk, Andrei; Gu, Allen; Bradley, Robert S.; Lu, Xuekun; Withers, Philip J.; Clarke, Amy; Henderson, Kevin; Cordes, Nikolaus; Patterson, Brian M.

    2015-09-01

    X-ray microscopy (XRM) has emerged as a powerful technique that reveals 3D images and quantitative information of interior structures. XRM executed both in the laboratory and at the synchrotron have demonstrated critical analysis and materials characterization on meso-, micro-, and nanoscales, with spatial resolution down to 50 nm in laboratory systems. The non-destructive nature of X-rays has made the technique widely appealing, with potential for "4D" characterization, delivering 3D micro- and nanostructural information on the same sample as a function of sequential processing or experimental conditions. Understanding volumetric and nanostructural changes, such as solid deformation, pore evolution, and crack propagation are fundamental to understanding how materials form, deform, and perform. We will present recent instrumentation developments in laboratory based XRM including a novel in situ nanomechanical testing stage. These developments bridge the gap between existing in situ stages for micro scale XRM, and SEM/TEM techniques that offer nanometer resolution but are limited to analysis of surfaces or extremely thin samples whose behavior is strongly influenced by surface effects. Several applications will be presented including 3D-characterization and in situ mechanical testing of polymers, metal alloys, composites and biomaterials. They span multiple length scales from the micro- to the nanoscale and different mechanical testing modes such as compression, indentation and tension.

  6. A one-piece 3D printed flexure translation stage for open-source microscopy.

    PubMed

    Sharkey, James P; Foo, Darryl C W; Kabla, Alexandre; Baumberg, Jeremy J; Bowman, Richard W

    2016-02-01

    Open source hardware has the potential to revolutionise the way we build scientific instruments; with the advent of readily available 3D printers, mechanical designs can now be shared, improved, and replicated faster and more easily than ever before. However, printed parts are typically plastic and often perform poorly compared to traditionally machined mechanisms. We have overcome many of the limitations of 3D printed mechanisms by exploiting the compliance of the plastic to produce a monolithic 3D printed flexure translation stage, capable of sub-micron-scale motion over a range of 8 × 8 × 4 mm. This requires minimal post-print clean-up and can be automated with readily available stepper motors. The resulting plastic composite structure is very stiff and exhibits remarkably low drift, moving less than 20 μm over the course of a week, without temperature stabilisation. This enables us to construct a miniature microscope with excellent mechanical stability, perfect for time-lapse measurements in situ in an incubator or fume hood. The ease of manufacture lends itself to use in containment facilities where disposability is advantageous and to experiments requiring many microscopes in parallel. High performance mechanisms based on printed flexures need not be limited to microscopy, and we anticipate their use in other devices both within the laboratory and beyond.

  7. A one-piece 3D printed flexure translation stage for open-source microscopy

    NASA Astrophysics Data System (ADS)

    Sharkey, James P.; Foo, Darryl C. W.; Kabla, Alexandre; Baumberg, Jeremy J.; Bowman, Richard W.

    2016-02-01

    Open source hardware has the potential to revolutionise the way we build scientific instruments; with the advent of readily available 3D printers, mechanical designs can now be shared, improved, and replicated faster and more easily than ever before. However, printed parts are typically plastic and often perform poorly compared to traditionally machined mechanisms. We have overcome many of the limitations of 3D printed mechanisms by exploiting the compliance of the plastic to produce a monolithic 3D printed flexure translation stage, capable of sub-micron-scale motion over a range of 8 × 8 × 4 mm. This requires minimal post-print clean-up and can be automated with readily available stepper motors. The resulting plastic composite structure is very stiff and exhibits remarkably low drift, moving less than 20 μm over the course of a week, without temperature stabilisation. This enables us to construct a miniature microscope with excellent mechanical stability, perfect for time-lapse measurements in situ in an incubator or fume hood. The ease of manufacture lends itself to use in containment facilities where disposability is advantageous and to experiments requiring many microscopes in parallel. High performance mechanisms based on printed flexures need not be limited to microscopy, and we anticipate their use in other devices both within the laboratory and beyond.

  8. A one-piece 3D printed flexure translation stage for open-source microscopy.

    PubMed

    Sharkey, James P; Foo, Darryl C W; Kabla, Alexandre; Baumberg, Jeremy J; Bowman, Richard W

    2016-02-01

    Open source hardware has the potential to revolutionise the way we build scientific instruments; with the advent of readily available 3D printers, mechanical designs can now be shared, improved, and replicated faster and more easily than ever before. However, printed parts are typically plastic and often perform poorly compared to traditionally machined mechanisms. We have overcome many of the limitations of 3D printed mechanisms by exploiting the compliance of the plastic to produce a monolithic 3D printed flexure translation stage, capable of sub-micron-scale motion over a range of 8 × 8 × 4 mm. This requires minimal post-print clean-up and can be automated with readily available stepper motors. The resulting plastic composite structure is very stiff and exhibits remarkably low drift, moving less than 20 μm over the course of a week, without temperature stabilisation. This enables us to construct a miniature microscope with excellent mechanical stability, perfect for time-lapse measurements in situ in an incubator or fume hood. The ease of manufacture lends itself to use in containment facilities where disposability is advantageous and to experiments requiring many microscopes in parallel. High performance mechanisms based on printed flexures need not be limited to microscopy, and we anticipate their use in other devices both within the laboratory and beyond. PMID:26931888

  9. 3D Imaging of Diatoms with Ion-abrasion Scanning Electron Microscopy

    PubMed Central

    Hildebrand, Mark; Kim, Sang; Shi, Dan; Scott, Keana; Subramaniam, Sriram

    2009-01-01

    Ion-abrasion scanning electron microscopy (IASEM) takes advantage of focused ion beams to abrade thin sections from the surface of bulk specimens, coupled with SEM to image the surface of each section, enabling 3D reconstructions of subcellular architecture at ~ 30 nm resolution. Here, we report the first application of IASEM for imaging a biomineralizing organism, the marine diatom Thalassiosira pseudonana. Diatoms have highly patterned silica-based cell wall structures that are unique models for the study and application of directed nanomaterials synthesis by biological systems. Our study provides new insights into the architecture and assembly principles of both the “hard” (siliceous) and “soft” (organic) components of the cell. From 3D reconstructions of developmentally synchronized diatoms captured at different stages, we show that both micro- and nanoscale siliceous structures can be visualized at specific stages in their formation. We show that not only are structures visualized in a whole-cell context, but demonstrate that fragile, early-stage structures are visible, and that this can be combined with elemental mapping in the exposed slice. We demonstrate that the 3D architectures of silica structures, and the cellular components that mediate their creation and positioning can be visualized simultaneously, providing new opportunities to study and manipulate mineral nanostructures in a genetically tractable system. PMID:19269330

  10. 3D motion of DNA-Au nanoconjugates in graphene liquid cell electron microscopy.

    PubMed

    Chen, Qian; Smith, Jessica M; Park, Jungwon; Kim, Kwanpyo; Ho, Davy; Rasool, Haider I; Zettl, Alex; Alivisatos, A Paul

    2013-09-11

    Liquid-phase transmission electron microscopy (TEM) can probe and visualize dynamic events with structural or functional details at the nanoscale in a liquid medium. Earlier efforts have focused on the growth and transformation kinetics of hard material systems, relying on their stability under electron beam. Our recently developed graphene liquid cell technique pushed the spatial resolution of such imaging to the atomic scale but still focused on growth trajectories of metallic nanocrystals. Here, we adopt this technique to imaging three-dimensional (3D) dynamics of soft materials instead, double strand (dsDNA) connecting Au nanocrystals as one example, at nanometer resolution. We demonstrate first that a graphene liquid cell can seal an aqueous sample solution of a lower vapor pressure than previously investigated well against the high vacuum in TEM. Then, from quantitative analysis of real time nanocrystal trajectories, we show that the status and configuration of dsDNA dictate the motions of linked nanocrystals throughout the imaging time of minutes. This sustained connecting ability of dsDNA enables this unprecedented continuous imaging of its dynamics via TEM. Furthermore, the inert graphene surface minimizes sample-substrate interaction and allows the whole nanostructure to rotate freely in the liquid environment; we thus develop and implement the reconstruction of 3D configuration and motions of the nanostructure from the series of 2D projected TEM images captured while it rotates. In addition to further proving the nanoconjugate structural stability, this reconstruction demonstrates 3D dynamic imaging by TEM beyond its conventional use in seeing a flattened and dry sample. Altogether, we foresee the new and exciting use of graphene liquid cell TEM in imaging 3D biomolecular transformations or interaction dynamics at nanometer resolution. PMID:23944844

  11. Optical turbulence above mountains seen in 3D

    NASA Astrophysics Data System (ADS)

    Els, S. G.; Vogiatzis, K.; Otárola, A.; Riddle, R.; Schöck, M.; Skidmore, W.; Travouillon, T.

    2010-07-01

    Atmospheric optical turbulence is the main driver of wavefront distortions which affect optical telescope performance. Therefore, many techniques have been developed to measure the optical turbulence strength along the line of sight. Based on data collected with the MASS (Multi Aperture Scintillation Sensor), we show that a large sample of such measurements can be used to assess the average three dimensional turbulence distribution above ground. The use of, and a more sophisticated instrumental setup for, such turbulence tomography will be discussed.

  12. Brightness-compensated 3-D optical flow algorithm for monitoring cochlear motion patterns

    NASA Astrophysics Data System (ADS)

    von Tiedemann, Miriam; Fridberger, Anders; Ulfendahl, Mats; de Monvel, Jacques Boutet

    2010-09-01

    A method for three-dimensional motion analysis designed for live cell imaging by fluorescence confocal microscopy is described. The approach is based on optical flow computation and takes into account brightness variations in the image scene that are not due to motion, such as photobleaching or fluorescence variations that may reflect changes in cellular physiology. The 3-D optical flow algorithm allowed almost perfect motion estimation on noise-free artificial sequences, and performed with a relative error of <10% on noisy images typical of real experiments. The method was applied to a series of 3-D confocal image stacks from an in vitro preparation of the guinea pig cochlea. The complex motions caused by slow pressure changes in the cochlear compartments were quantified. At the surface of the hearing organ, the largest motion component was the transverse one (normal to the surface), but significant radial and longitudinal displacements were also present. The outer hair cell displayed larger radial motion at their basolateral membrane than at their apical surface. These movements reflect mechanical interactions between different cellular structures, which may be important for communicating sound-evoked vibrations to the sensory cells. A better understanding of these interactions is important for testing realistic models of cochlear mechanics.

  13. Test target for characterizing 3D resolution of optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Hu, Zhixiong; Hao, Bingtao; Liu, Wenli; Hong, Baoyu; Li, Jiao

    2014-12-01

    Optical coherence tomography (OCT) is a non-invasive 3D imaging technology which has been applied or investigated in many diagnostic fields including ophthalmology, dermatology, dentistry, cardiovasology, endoscopy, brain imaging and so on. Optical resolution is an important characteristic that can describe the quality and utility of an image acquiring system. We employ 3D printing technology to design and fabricate a test target for characterizing 3D resolution of optical coherence tomography. The test target which mimics USAF 1951 test chart was produced with photopolymer. By measuring the 3D test target, axial resolution as well as lateral resolution of a spectral domain OCT system was evaluated. For comparison, conventional microscope and surface profiler were employed to characterize the 3D test targets. The results demonstrate that the 3D resolution test targets have the potential of qualitatively and quantitatively validating the performance of OCT systems.

  14. Focusing optics of a parallel beam CCD optical tomography apparatus for 3D radiation gel dosimetry.

    PubMed

    Krstajić, Nikola; Doran, Simon J

    2006-04-21

    Optical tomography of gel dosimeters is a promising and cost-effective avenue for quality control of radiotherapy treatments such as intensity-modulated radiotherapy (IMRT). Systems based on a laser coupled to a photodiode have so far shown the best results within the context of optical scanning of radiosensitive gels, but are very slow ( approximately 9 min per slice) and poorly suited to measurements that require many slices. Here, we describe a fast, three-dimensional (3D) optical computed tomography (optical-CT) apparatus, based on a broad, collimated beam, obtained from a high power LED and detected by a charged coupled detector (CCD). The main advantages of such a system are (i) an acquisition speed approximately two orders of magnitude higher than a laser-based system when 3D data are required, and (ii) a greater simplicity of design. This paper advances our previous work by introducing a new design of focusing optics, which take information from a suitably positioned focal plane and project an image onto the CCD. An analysis of the ray optics is presented, which explains the roles of telecentricity, focusing, acceptance angle and depth-of-field (DOF) in the formation of projections. A discussion of the approximation involved in measuring the line integrals required for filtered backprojection reconstruction is given. Experimental results demonstrate (i) the effect on projections of changing the position of the focal plane of the apparatus, (ii) how to measure the acceptance angle of the optics, and (iii) the ability of the new scanner to image both absorbing and scattering gel phantoms. The quality of reconstructed images is very promising and suggests that the new apparatus may be useful in a clinical setting for fast and accurate 3D dosimetry. PMID:16585845

  15. Focusing optics of a parallel beam CCD optical tomography apparatus for 3D radiation gel dosimetry

    NASA Astrophysics Data System (ADS)

    Krstajic, Nikola; Doran, Simon J.

    2006-04-01

    Optical tomography of gel dosimeters is a promising and cost-effective avenue for quality control of radiotherapy treatments such as intensity-modulated radiotherapy (IMRT). Systems based on a laser coupled to a photodiode have so far shown the best results within the context of optical scanning of radiosensitive gels, but are very slow (~9 min per slice) and poorly suited to measurements that require many slices. Here, we describe a fast, three-dimensional (3D) optical computed tomography (optical-CT) apparatus, based on a broad, collimated beam, obtained from a high power LED and detected by a charged coupled detector (CCD). The main advantages of such a system are (i) an acquisition speed approximately two orders of magnitude higher than a laser-based system when 3D data are required, and (ii) a greater simplicity of design. This paper advances our previous work by introducing a new design of focusing optics, which take information from a suitably positioned focal plane and project an image onto the CCD. An analysis of the ray optics is presented, which explains the roles of telecentricity, focusing, acceptance angle and depth-of-field (DOF) in the formation of projections. A discussion of the approximation involved in measuring the line integrals required for filtered backprojection reconstruction is given. Experimental results demonstrate (i) the effect on projections of changing the position of the focal plane of the apparatus, (ii) how to measure the acceptance angle of the optics, and (iii) the ability of the new scanner to image both absorbing and scattering gel phantoms. The quality of reconstructed images is very promising and suggests that the new apparatus may be useful in a clinical setting for fast and accurate 3D dosimetry.

  16. 3D elemental sensitive imaging using transmission X-ray microscopy.

    PubMed

    Liu, Yijin; Meirer, Florian; Wang, Junyue; Requena, Guillermo; Williams, Phillip; Nelson, Johanna; Mehta, Apurva; Andrews, Joy C; Pianetta, Piero

    2012-09-01

    Determination of the heterogeneous distribution of metals in alloy/battery/catalyst and biological materials is critical to fully characterize and/or evaluate the functionality of the materials. Using synchrotron-based transmission x-ray microscopy (TXM), it is now feasible to perform nanoscale-resolution imaging over a wide X-ray energy range covering the absorption edges of many elements; combining elemental sensitive imaging with determination of sample morphology. We present an efficient and reliable methodology to perform 3D elemental sensitive imaging with excellent sample penetration (tens of microns) using hard X-ray TXM. A sample of an Al-Si piston alloy is used to demonstrate the capability of the proposed method. PMID:22349401

  17. Electron Microscopy: From 2D to 3D Images with Special Reference to Muscle

    PubMed Central

    2015-01-01

    This is a brief and necessarily very sketchy presentation of the evolution in electron microscopy (EM) imaging that was driven by the necessity of extracting 3-D views from the essentially 2-D images produced by the electron beam. The lens design of standard transmission electron microscope has not been greatly altered since its inception. However, technical advances in specimen preparation, image collection and analysis gradually induced an astounding progression over a period of about 50 years. From the early images that redefined tissues, cell and cell organelles at the sub-micron level, to the current nano-resolution reconstructions of organelles and proteins the step is very large. The review is written by an investigator who has followed the field for many years, but often from the sidelines, and with great wonder. Her interest in muscle ultrastructure colors the writing. More specific detailed reviews are presented in this issue. PMID:26913146

  18. 3D elemental sensitive imaging using transmission X-ray microscopy.

    PubMed

    Liu, Yijin; Meirer, Florian; Wang, Junyue; Requena, Guillermo; Williams, Phillip; Nelson, Johanna; Mehta, Apurva; Andrews, Joy C; Pianetta, Piero

    2012-09-01

    Determination of the heterogeneous distribution of metals in alloy/battery/catalyst and biological materials is critical to fully characterize and/or evaluate the functionality of the materials. Using synchrotron-based transmission x-ray microscopy (TXM), it is now feasible to perform nanoscale-resolution imaging over a wide X-ray energy range covering the absorption edges of many elements; combining elemental sensitive imaging with determination of sample morphology. We present an efficient and reliable methodology to perform 3D elemental sensitive imaging with excellent sample penetration (tens of microns) using hard X-ray TXM. A sample of an Al-Si piston alloy is used to demonstrate the capability of the proposed method.

  19. The Use of Atomic Force Microscopy for 3D Analysis of Nucleic Acid Hybridization on Microarrays

    PubMed Central

    Dubrovin, E. V.; Presnova, G. V.; Rubtsova, M. Yu.; Egorov, A. M.; Grigorenko, V. G.; Yaminsky, I. V.

    2015-01-01

    Oligonucleotide microarrays are considered today to be one of the most efficient methods of gene diagnostics. The capability of atomic force microscopy (AFM) to characterize the three-dimensional morphology of single molecules on a surface allows one to use it as an effective tool for the 3D analysis of a microarray for the detection of nucleic acids. The high resolution of AFM offers ways to decrease the detection threshold of target DNA and increase the signal-to-noise ratio. In this work, we suggest an approach to the evaluation of the results of hybridization of gold nanoparticle-labeled nucleic acids on silicon microarrays based on an AFM analysis of the surface both in air and in liquid which takes into account of their three-dimensional structure. We suggest a quantitative measure of the hybridization results which is based on the fraction of the surface area occupied by the nanoparticles. PMID:26085952

  20. Electron Microscopy: From 2D to 3D Images with Special Reference to Muscle.

    PubMed

    Franzini-Armstrong, Clara

    2015-01-01

    This is a brief and necessarily very sketchy presentation of the evolution in electron microscopy (EM) imaging that was driven by the necessity of extracting 3-D views from the essentially 2-D images produced by the electron beam. The lens design of standard transmission electron microscope has not been greatly altered since its inception. However, technical advances in specimen preparation, image collection and analysis gradually induced an astounding progression over a period of about 50 years. From the early images that redefined tissues, cell and cell organelles at the sub-micron level, to the current nano-resolution reconstructions of organelles and proteins the step is very large. The review is written by an investigator who has followed the field for many years, but often from the sidelines, and with great wonder. Her interest in muscle ultrastructure colors the writing. More specific detailed reviews are presented in this issue. PMID:26913146

  1. Precise quantification of silica and ceria nanoparticle uptake revealed by 3D fluorescence microscopy

    PubMed Central

    Torrano, Adriano A

    2014-01-01

    Summary Particle_in_Cell-3D is a powerful method to quantify the cellular uptake of nanoparticles. It combines the advantages of confocal fluorescence microscopy with fast and precise semi-automatic image analysis. In this work we present how this method was applied to investigate the impact of 310 nm silica nanoparticles on human vascular endothelial cells (HUVEC) in comparison to a cancer cell line derived from the cervix carcinoma (HeLa). The absolute number of intracellular silica nanoparticles within the first 24 h was determined and shown to be cell type-dependent. As a second case study, Particle_in_Cell-3D was used to assess the uptake kinetics of 8 nm and 30 nm ceria nanoparticles interacting with human microvascular endothelial cells (HMEC-1). These small nanoparticles formed agglomerates in biological medium, and the particles that were in effective contact with cells had a mean diameter of 417 nm and 316 nm, respectively. A significant particle size-dependent effect was observed after 48 h of interaction, and the number of intracellular particles was more than four times larger for the 316 nm agglomerates. Interestingly, our results show that for both particle sizes there is a maximum dose of intracellular nanoparticles at about 24 h. One of the causes for such an interesting and unusual uptake behavior could be cell division. PMID:25383274

  2. 3D surface reconstruction and FIB microscopy of worn alumina hip prostheses

    NASA Astrophysics Data System (ADS)

    Zeng, P.; Inkson, B. J.; Rainforth, W. M.; Stewart, T.

    2008-08-01

    Interest in alumina-on-alumina total hip replacements (THR) continues to grow for the young and active patient due to their superior wear performance and biocompatibility compared to the alternative traditional polymer/metal prostheses. While alumina on alumina bearings offer an excellent solution, a region of high wear, known as stripe wear, is commonly observed on retrieved alumina hip components that poses concern. These in-vivo stripe wear mechanisms can be replicated in vitro by the introduction of micro-separation during the simulated walking cycle in hip joint simulation. However, the understanding of the mechanisms behind the stripe wear processes is relatively poor. 3D topographic reconstructions of titled SEM stereo pairs from different zones have been obtained to determine the local worn surface topography. Focused ion beam (FIB) microscopy was applied to examine the subsurface damage across the stripe wear. The paper presents novel images of sub-surface microcracks in alumina along with 3D reconstructions of the worn ceramic surfaces and a classification of four distinct wear zones following microseparation in hip prostheses.

  3. Local characterization of hindered Brownian motion by using digital video microscopy and 3D particle tracking

    SciTech Connect

    Dettmer, Simon L.; Keyser, Ulrich F.; Pagliara, Stefano

    2014-02-15

    In this article we present methods for measuring hindered Brownian motion in the confinement of complex 3D geometries using digital video microscopy. Here we discuss essential features of automated 3D particle tracking as well as diffusion data analysis. By introducing local mean squared displacement-vs-time curves, we are able to simultaneously measure the spatial dependence of diffusion coefficients, tracking accuracies and drift velocities. Such local measurements allow a more detailed and appropriate description of strongly heterogeneous systems as opposed to global measurements. Finite size effects of the tracking region on measuring mean squared displacements are also discussed. The use of these methods was crucial for the measurement of the diffusive behavior of spherical polystyrene particles (505 nm diameter) in a microfluidic chip. The particles explored an array of parallel channels with different cross sections as well as the bulk reservoirs. For this experiment we present the measurement of local tracking accuracies in all three axial directions as well as the diffusivity parallel to the channel axis while we observed no significant flow but purely Brownian motion. Finally, the presented algorithm is suitable also for tracking of fluorescently labeled particles and particles driven by an external force, e.g., electrokinetic or dielectrophoretic forces.

  4. Computational 3D reconstructions by optimization for cryo-electron microscopy

    NASA Astrophysics Data System (ADS)

    Yin, Zhye; Zheng, Yili; Doerschuk, Peter C.; Johnson, John E.

    2003-06-01

    An algorithm for the simultaneous 3-D reconstruction of several types of object, where each type of object may possibly have a rotational symmetry, from 2-D projection images, where for each image the type of object imaged, the projection orientation used to create the image, and the location of the object in the image are unknown, is described. The motivating application is the determination of the 3-D structure of small spherical viruses from cryo electron microscopy images. The algorithm is a maximum likelihood estimator which is computed by expectation maximization (EM). Due to the structure of the statistical model, the maximization step of EM can be easily computed but the expectation step requires 5-D numerical quadrature. The computational burden of the quadratures necessitates parallel computation and three different implementations of two different types of parallelism have been developed using pthreads (for shared memory processors) and MPI (for distributed memory processors). An example applying one of the MPI implementations, running on a 32 node PC cluster, to experimental images of Flock House Virus with comparison to the x-ray crystal diffraction structure of the virus is described.

  5. Super-resolution imaging of the cytokinetic Z ring in live bacteria using fast 3D-structured illumination microscopy (f3D-SIM).

    PubMed

    Turnbull, Lynne; Strauss, Michael P; Liew, Andrew T F; Monahan, Leigh G; Whitchurch, Cynthia B; Harry, Elizabeth J

    2014-01-01

    Imaging of biological samples using fluorescence microscopy has advanced substantially with new technologies to overcome the resolution barrier of the diffraction of light allowing super-resolution of live samples. There are currently three main types of super-resolution techniques - stimulated emission depletion (STED), single-molecule localization microscopy (including techniques such as PALM, STORM, and GDSIM), and structured illumination microscopy (SIM). While STED and single-molecule localization techniques show the largest increases in resolution, they have been slower to offer increased speeds of image acquisition. Three-dimensional SIM (3D-SIM) is a wide-field fluorescence microscopy technique that offers a number of advantages over both single-molecule localization and STED. Resolution is improved, with typical lateral and axial resolutions of 110 and 280 nm, respectively and depth of sampling of up to 30 µm from the coverslip, allowing for imaging of whole cells. Recent advancements (fast 3D-SIM) in the technology increasing the capture rate of raw images allows for fast capture of biological processes occurring in seconds, while significantly reducing photo-toxicity and photobleaching. Here we describe the use of one such method to image bacterial cells harboring the fluorescently-labelled cytokinetic FtsZ protein to show how cells are analyzed and the type of unique information that this technique can provide.

  6. Super-resolution imaging of the cytokinetic Z ring in live bacteria using fast 3D-structured illumination microscopy (f3D-SIM).

    PubMed

    Turnbull, Lynne; Strauss, Michael P; Liew, Andrew T F; Monahan, Leigh G; Whitchurch, Cynthia B; Harry, Elizabeth J

    2014-01-01

    Imaging of biological samples using fluorescence microscopy has advanced substantially with new technologies to overcome the resolution barrier of the diffraction of light allowing super-resolution of live samples. There are currently three main types of super-resolution techniques - stimulated emission depletion (STED), single-molecule localization microscopy (including techniques such as PALM, STORM, and GDSIM), and structured illumination microscopy (SIM). While STED and single-molecule localization techniques show the largest increases in resolution, they have been slower to offer increased speeds of image acquisition. Three-dimensional SIM (3D-SIM) is a wide-field fluorescence microscopy technique that offers a number of advantages over both single-molecule localization and STED. Resolution is improved, with typical lateral and axial resolutions of 110 and 280 nm, respectively and depth of sampling of up to 30 µm from the coverslip, allowing for imaging of whole cells. Recent advancements (fast 3D-SIM) in the technology increasing the capture rate of raw images allows for fast capture of biological processes occurring in seconds, while significantly reducing photo-toxicity and photobleaching. Here we describe the use of one such method to image bacterial cells harboring the fluorescently-labelled cytokinetic FtsZ protein to show how cells are analyzed and the type of unique information that this technique can provide. PMID:25286090

  7. Spectral fusing Gabor domain optical coherence microscopy.

    PubMed

    Meemon, Panomsak; Widjaja, Joewono; Rolland, Jannick P

    2016-02-01

    Gabor domain optical coherence microscopy (GD-OCM) is one of many variations of optical coherence tomography (OCT) techniques that aims for invariant high resolution across a 3D field of view by utilizing the ability to dynamically refocus the imaging optics in the sample arm. GD-OCM acquires multiple cross-sectional images at different focus positions of the objective lens, and then fuses them to obtain an invariant high-resolution 3D image of the sample, which comes with the intrinsic drawback of a longer processing time as compared to conventional Fourier domain OCT. Here, we report on an alternative Gabor fusing algorithm, the spectral-fusion technique, which directly processes each acquired spectrum and combines them prior to the Fourier transformation to obtain a depth profile. The implementation of the spectral-fusion algorithm is presented and its performance is compared to that of the prior GD-OCM spatial-fusion approach. The spectral-fusion approach shows twice the speed of the spatial-fusion approach for a spectrum size of less than 2000 point sampling, which is a commonly used spectrum size in OCT imaging, including GD-OCM.

  8. Fast segmentation of stained nuclei in terabyte-scale, time resolved 3D microscopy image stacks.

    PubMed

    Stegmaier, Johannes; Otte, Jens C; Kobitski, Andrei; Bartschat, Andreas; Garcia, Ariel; Nienhaus, G Ulrich; Strähle, Uwe; Mikut, Ralf

    2014-01-01

    Automated analysis of multi-dimensional microscopy images has become an integral part of modern research in life science. Most available algorithms that provide sufficient segmentation quality, however, are infeasible for a large amount of data due to their high complexity. In this contribution we present a fast parallelized segmentation method that is especially suited for the extraction of stained nuclei from microscopy images, e.g., of developing zebrafish embryos. The idea is to transform the input image based on gradient and normal directions in the proximity of detected seed points such that it can be handled by straightforward global thresholding like Otsu's method. We evaluate the quality of the obtained segmentation results on a set of real and simulated benchmark images in 2D and 3D and show the algorithm's superior performance compared to other state-of-the-art algorithms. We achieve an up to ten-fold decrease in processing times, allowing us to process large data sets while still providing reasonable segmentation results.

  9. In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Lee, Seunghun; Lee, Jun Ho; Park, Jin Hyoung; Yoon, Yeoreum; Chung, Wan Kyun; Tchah, Hungwon; Kim, Myoung Joon; Kim, Ki Hean

    2016-05-01

    Moxifloxacin and gatifloxacin are fourth-generation fluoroquinolone antibiotics used in the clinic to prevent or treat ocular infections. Their pharmacokinetics in the cornea is usually measured from extracted ocular fluids or tissues, and in vivo direct measurement is difficult. In this study multiphoton microscopy (MPM), which is a 3D optical microscopic technique based on multiphoton fluorescence, was applied to the measurement of moxifloxacin and gatifloxacin distribution in the cornea. Intrinsic multiphoton fluorescence properties of moxifloxacin and gatifloxacin were characterized, and their distributions in mouse cornea in vivo were measured by 3D MPM imaging. Both moxifloxacin and gatifloxacin had similar multiphoton spectra, while moxifloxacin had stronger fluorescence than gatifloxacin. MPM imaging of mouse cornea in vivo showed (1) moxifloxacin had good penetration through the superficial corneal epithelium, while gatifloxacin had relatively poor penetration, (2) both ophthalmic solutions had high intracellular distribution. In vivo MPM results were consistent with previous studies. This study demonstrates the feasibility of MPM as a method for in vivo direct measurement of moxifloxacin and gatifloxacin in the cornea.

  10. Holographic microscopy and microfluidics platform for measuring wall stress and 3D flow over surfaces textured by micro-pillars

    PubMed Central

    Bocanegra Evans, Humberto; Gorumlu, Serdar; Aksak, Burak; Castillo, Luciano; Sheng, Jian

    2016-01-01

    Understanding how fluid flow interacts with micro-textured surfaces is crucial for a broad range of key biological processes and engineering applications including particle dispersion, pathogenic infections, and drag manipulation by surface topology. We use high-speed digital holographic microscopy (DHM) in combination with a correlation based de-noising algorithm to overcome the optical interference generated by surface roughness and to capture a large number of 3D particle trajectories in a microfluidic channel with one surface patterned with micropillars. It allows us to obtain a 3D ensembled velocity field with an uncertainty of 0.06% and 2D wall shear stress distribution at the resolution of ~65 μPa. Contrary to laminar flow in most microfluidics, we find that the flow is three-dimensional and complex for the textured microchannel. While the micropillars affect the velocity flow field locally, their presence is felt globally in terms of wall shear stresses at the channel walls. These findings imply that micro-scale mixing and wall stress sensing/manipulation can be achieved through hydro-dynamically smooth but topologically rough micropillars. PMID:27353632

  11. Holographic microscopy and microfluidics platform for measuring wall stress and 3D flow over surfaces textured by micro-pillars

    NASA Astrophysics Data System (ADS)

    Bocanegra Evans, Humberto; Gorumlu, Serdar; Aksak, Burak; Castillo, Luciano; Sheng, Jian

    2016-06-01

    Understanding how fluid flow interacts with micro-textured surfaces is crucial for a broad range of key biological processes and engineering applications including particle dispersion, pathogenic infections, and drag manipulation by surface topology. We use high-speed digital holographic microscopy (DHM) in combination with a correlation based de-noising algorithm to overcome the optical interference generated by surface roughness and to capture a large number of 3D particle trajectories in a microfluidic channel with one surface patterned with micropillars. It allows us to obtain a 3D ensembled velocity field with an uncertainty of 0.06% and 2D wall shear stress distribution at the resolution of ~65 μPa. Contrary to laminar flow in most microfluidics, we find that the flow is three-dimensional and complex for the textured microchannel. While the micropillars affect the velocity flow field locally, their presence is felt globally in terms of wall shear stresses at the channel walls. These findings imply that micro-scale mixing and wall stress sensing/manipulation can be achieved through hydro-dynamically smooth but topologically rough micropillars.

  12. 3-D Adaptive Sparsity Based Image Compression with Applications to Optical Coherence Tomography

    PubMed Central

    Fang, Leyuan; Li, Shutao; Kang, Xudong; Izatt, Joseph A.; Farsiu, Sina

    2015-01-01

    We present a novel general-purpose compression method for tomographic images, termed 3D adaptive sparse representation based compression (3D-ASRC). In this paper, we focus on applications of 3D-ASRC for the compression of ophthalmic 3D optical coherence tomography (OCT) images. The 3D-ASRC algorithm exploits correlations among adjacent OCT images to improve compression performance, yet is sensitive to preserving their differences. Due to the inherent denoising mechanism of the sparsity based 3D-ASRC, the quality of the compressed images are often better than the raw images they are based on. Experiments on clinical-grade retinal OCT images demonstrate the superiority of the proposed 3D-ASRC over other well-known compression methods. PMID:25561591

  13. Neutron detection and characterization for non-proliferation applications using 3D computer optical memories [Use of 3D optical computer memory for radiation detectors/dosimeters. Final progress report

    SciTech Connect

    Gary W. Phillips

    2000-12-20

    We have investigated 3-dimensional optical random access memory (3D-ORAM) materials for detection and characterization of charged particles of neutrons by detecting tracks left by the recoil charged particles produced by the neutrons. We have characterized the response of these materials to protons, alpha particles and carbon-12 nuclei as a functions of dose and energy. We have observed individual tracks using scanning electron microscopy and atomic force microscopy. We are investigating the use of neural net analysis to characterize energetic neutron fields from their track structure in these materials.

  14. Light sheet adaptive optics microscope for 3D live imaging

    NASA Astrophysics Data System (ADS)

    Bourgenot, C.; Taylor, J. M.; Saunter, C. D.; Girkin, J. M.; Love, G. D.

    2013-02-01

    We report on the incorporation of adaptive optics (AO) into the imaging arm of a selective plane illumination microscope (SPIM). SPIM has recently emerged as an important tool for life science research due to its ability to deliver high-speed, optically sectioned, time-lapse microscope images from deep within in vivo selected samples. SPIM provides a very interesting system for the incorporation of AO as the illumination and imaging paths are decoupled and AO may be useful in both paths. In this paper, we will report the use of AO applied to the imaging path of a SPIM, demonstrating significant improvement in image quality of a live GFP-labeled transgenic zebrafish embryo heart using a modal, wavefront sensorless approach and a heart synchronization method. These experimental results are linked to a computational model showing that significant aberrations are produced by the tube holding the sample in addition to the aberration from the biological sample itself.

  15. Towards a 3-D Magneto-Optical Trap for Molecules

    NASA Astrophysics Data System (ADS)

    Collopy, Alejandra; Hummon, Matthew; Yeo, Mark; Stuhl, Benjamin; Hemmerling, Boerge; Drayna, Garrett; Chae, Eunmi; Ravi, Aakash; Lu, Hsin-I.; Doyle, John; Ye, Jun

    2013-05-01

    As the magneto-optical trap revolutionized atomic physics, we anticipate the molecular counterpart to open doors to unexplored molecular physics, including ultra-cold chemistry. While molecules possess more complex structure than atoms, quasi-cycling cooling transitions are still attainable in a variety of species, including the polar molecule YO. In order to remix dark states, we RF modulate the polarization of the light in our trap. In order to maintain a restoring force, we modulate the orientation of our magnetic fields in phase with the light using LC resonant in-vacuum magnetic coils. We demonstrate magneto-optical trapping in two dimensions for YO, and present progress towards a three dimensional implementation of a MOT loaded from a two-stage buffer gas cell source. We acknowledge support from the AFOSR (MURI), DOE, NIST and the NSF.

  16. Subtractive 3D printing of optically active diamond structures.

    PubMed

    Martin, Aiden A; Toth, Milos; Aharonovich, Igor

    2014-05-21

    Controlled fabrication of semiconductor nanostructures is an essential step in engineering of high performance photonic and optoelectronic devices. Diamond in particular has recently attracted considerable attention as a promising platform for quantum technologies, photonics and high resolution sensing applications. Here we demonstrate the fabrication of optically active, functional diamond structures using gas-mediated electron beam induced etching (EBIE). The technique achieves dry chemical etching at room temperature through the dissociation of surface-adsorbed H2O molecules by energetic electrons in a water vapor environment. Parallel processing is possible by electron flood exposure and the use of an etch mask, while high resolution, mask-free, iterative editing is demonstrated by direct write etching of inclined facets of diamond microparticles. The realized structures demonstrate the potential of EBIE for the fabrication of optically active structures in diamond.

  17. The potential of 3D-FISH and super-resolution structured illumination microscopy for studies of 3D nuclear architecture: 3D structured illumination microscopy of defined chromosomal structures visualized by 3D (immuno)-FISH opens new perspectives for studies of nuclear architecture.

    PubMed

    Markaki, Yolanda; Smeets, Daniel; Fiedler, Susanne; Schmid, Volker J; Schermelleh, Lothar; Cremer, Thomas; Cremer, Marion

    2012-05-01

    Three-dimensional structured illumination microscopy (3D-SIM) has opened up new possibilities to study nuclear architecture at the ultrastructural level down to the ~100 nm range. We present first results and assess the potential using 3D-SIM in combination with 3D fluorescence in situ hybridization (3D-FISH) for the topographical analysis of defined nuclear targets. Our study also deals with the concern that artifacts produced by FISH may counteract the gain in resolution. We address the topography of DAPI-stained DNA in nuclei before and after 3D-FISH, nuclear pores and the lamina, chromosome territories, chromatin domains, and individual gene loci. We also look at the replication patterns of chromocenters and the topographical relationship of Xist-RNA within the inactive X-territory. These examples demonstrate that an appropriately adapted 3D-FISH/3D-SIM approach preserves key characteristics of the nuclear ultrastructure and that the gain in information obtained by 3D-SIM yields new insights into the functional nuclear organization. PMID:22508100

  18. TRAIL protein localization in human primary T cells by 3D microscopy using 3D interactive surface plot: a new method to visualize plasma membrane.

    PubMed

    Gras, Christophe; Smith, Nikaïa; Sengmanivong, Lucie; Gandini, Mariana; Kubelka, Claire Fernandes; Herbeuval, Jean-Philippe

    2013-01-31

    The apoptotic ligand TNF-related apoptosis ligand (TRAIL) is expressed on the membrane of immune cells during HIV infection. The intracellular stockade of TRAIL in human primary CD4(+) T cells is not known. Here we investigated whether primary CD4(+) T cells expressed TRAIL in their intracellular compartment and whether TRAIL is relocalized on the plasma membrane under HIV activation. We found that TRAIL protein was stocked in intracellular compartment in non activated CD4(+) T cells and that the total level of TRAIL protein was not increased under HIV-1 stimulation. However, TRAIL was massively relocalized on plasma membrane when cells were cultured with HIV. Using three dimensional (3D) microscopy we localized TRAIL protein in human T cells and developed a new method to visualize plasma membrane without the need of a membrane marker. This method used the 3D interactive surface plot and bright light acquired images. PMID:23085529

  19. TRAIL protein localization in human primary T cells by 3D microscopy using 3D interactive surface plot: a new method to visualize plasma membrane.

    PubMed

    Gras, Christophe; Smith, Nikaïa; Sengmanivong, Lucie; Gandini, Mariana; Kubelka, Claire Fernandes; Herbeuval, Jean-Philippe

    2013-01-31

    The apoptotic ligand TNF-related apoptosis ligand (TRAIL) is expressed on the membrane of immune cells during HIV infection. The intracellular stockade of TRAIL in human primary CD4(+) T cells is not known. Here we investigated whether primary CD4(+) T cells expressed TRAIL in their intracellular compartment and whether TRAIL is relocalized on the plasma membrane under HIV activation. We found that TRAIL protein was stocked in intracellular compartment in non activated CD4(+) T cells and that the total level of TRAIL protein was not increased under HIV-1 stimulation. However, TRAIL was massively relocalized on plasma membrane when cells were cultured with HIV. Using three dimensional (3D) microscopy we localized TRAIL protein in human T cells and developed a new method to visualize plasma membrane without the need of a membrane marker. This method used the 3D interactive surface plot and bright light acquired images.

  20. 3D parameter reconstruction in hyperspectral diffuse optical tomography

    NASA Astrophysics Data System (ADS)

    Saibaba, Arvind K.; Krishnamurthy, Nishanth; Anderson, Pamela G.; Kainerstorfer, Jana M.; Sassaroli, Angelo; Miller, Eric L.; Fantini, Sergio; Kilmer, Misha E.

    2015-03-01

    The imaging of shape perturbation and chromophore concentration using Diffuse Optical Tomography (DOT) data can be mathematically described as an ill-posed and non-linear inverse problem. The reconstruction algorithm for hyperspectral data using a linearized Born model is prohibitively expensive, both in terms of computation and memory. We model the shape of the perturbation using parametric level-set approach (PaLS). We discuss novel computational strategies for reducing the computational cost based on a Krylov subspace approach for parameteric linear systems and a compression strategy for the parameter-to-observation map. We will demonstrate the validity of our approach by comparison with experiments.

  1. Snapshot 3D optical coherence tomography system using image mappingspectrometry

    PubMed Central

    Nguyen, Thuc-Uyen; Pierce, Mark C; Higgins, Laura; Tkaczyk, Tomasz S

    2013-01-01

    A snapshot 3-Dimensional Optical Coherence Tomography system was developed using Image MappingSpectrometry. This system can give depth information (Z) at different spatial positions (XY) withinone camera integration time to potentially reduce motion artifact and enhance throughput. Thecurrent (x,y,λ) datacube of (85×356×117) provides a 3Dvisualization of sample with 400 μm depth and 13.4μm in transverse resolution. Axial resolution of 16.0μm can also be achieved in this proof-of-concept system. We present ananalysis of the theoretical constraints which will guide development of future systems withincreased imaging depth and improved axial and lateral resolutions. PMID:23736629

  2. Controllable 3D atomic Brownian motor in optical lattices

    NASA Astrophysics Data System (ADS)

    Dion, C. M.; Sjölund, P.; Petra, S. J. H.; Jonsell, S.; Nylén, M.; Sanchez-Palencia, L.; Kastberg, A.

    2008-06-01

    We study a Brownian motor, based on cold atoms in optical lattices, where atomic motion can be induced in a controlled manner in an arbitrary direction, by rectification of isotropic random fluctuations. In contrast with ratchet mechanisms, our Brownian motor operates in a potential that is spatially and temporally symmetric, in apparent contradiction to the Curie principle. Simulations, based on the Fokker-Planck equation, allow us to gain knowledge on the qualitative behaviour of our Brownian motor. Studies of Brownian motors, and in particular ones with unique control properties, are of fundamental interest because of the role they play in protein motors and their potential applications in nanotechnology. In particular, our system opens the way to the study of quantum Brownian motors.

  3. 3D Printing Optical Engine for Controlling Material Microstructure

    NASA Astrophysics Data System (ADS)

    Huang, Wei-Chin; Chang, Kuang-Po; Wu, Ping-Han; Wu, Chih-Hsien; Lin, Ching-Chih; Chuang, Chuan-Sheng; Lin, De-Yau; Liu, Sung-Ho; Horng, Ji-Bin; Tsau, Fang-Hei

    Controlling the cooling rate of alloy during melting and resolidification is the most commonly used method for varying the material microstructure and consequently the resuling property. However, the cooling rate of a selective laser melting (SLM) production is restricted by a preset optimal parameter of a good dense product. The head room for locally manipulating material property in a process is marginal. In this study, we invent an Optical Engine for locally controlling material microstructure in a SLM process. It develops an invovative method to control and adjust thermal history of the solidification process to gain desired material microstucture and consequently drastically improving the quality. Process parameters selected locally for specific materials requirement according to designed characteristics by using thermal dynamic principles of solidification process. It utilize a technique of complex laser beam shape of adaptive irradiation profile to permit local control of material characteristics as desired. This technology could be useful for industrial application of medical implant, aerospace and automobile industries.

  4. An optical real-time 3D measurement for analysis of facial shape and movement

    NASA Astrophysics Data System (ADS)

    Zhang, Qican; Su, Xianyu; Chen, Wenjing; Cao, Yiping; Xiang, Liqun

    2003-12-01

    Optical non-contact 3-D shape measurement provides a novel and useful tool for analysis of facial shape and movement in presurgical and postsurgical regular check. In this article we present a system, which allows a precise 3-D visualization of the patient's facial before and after craniofacial surgery. We discussed, in this paper, the real time 3-D image capture, processing and the 3-D phase unwrapping method to recover complex shape deformation when the movement of the mouth. The result of real-time measurement for facial shape and movement will be helpful for the more ideal effect in plastic surgery.

  5. 3D measurements of live cells via digital holographic microscopy and terahertz spectroscopy

    NASA Astrophysics Data System (ADS)

    Park, Jun Yong; Oser, Dorian; Iapozzuto, Peter; Norbury, Sean; Mahajan, Supriya; Khmaladze, Alexander; Sharikova, Anna

    2016-03-01

    This is a study of the central nervous system (CNS) cells, including brain micro vascular endothelial cells (BMV) that constitute the blood brain barrier, and C6 glial cells that are the predominant cell in the brain. The cells are exposed to various chemicals by non-invasive, label-free methods. Digital holographic microscopy (DHM) is a technique that records an interference pattern between an object and reference waves, so that the computationally reconstructed holographic image contains both amplitude and phase information, and 3D images are obtained. The measurement of cell cultures by digital holographic microscopy yields information about cell death mechanisms, since these processes are correlated with individual cell volume. Our in-house DHM combines a visible (red) laser source with a conventional microscope base, and LabVIEW-run data processing. Terahertz spectral signatures are associated with structural changes in molecules and provide complementary information about cells. Both CNS cells BMV and C6 cells are treated with the drug "Methamphetamine" (METH), which induces apoptosis in neuronal cells and exhibits decrease in cell volume, a characteristic of cells undergoing apoptosis (induced cell death). METH can cause CNS cell death by cross-talk between mitochondria-, endoplasmic reticulum-, and receptor-mediated apoptotic events, all of which results in drug induced changes in neuroplasticity and significant neuropathology. Doxorubicin (DOX), a popular anticancer drug, is used as a control. We observe that METH treatment resulted in more pronounced cell volume shrinkage in both the BMV and C6 cells, as compared to DOX-induced cell apoptosis.

  6. Investigation of resins suitable for the preparation of biological sample for 3-D electron microscopy.

    PubMed

    Kizilyaprak, Caroline; Longo, Giovanni; Daraspe, Jean; Humbel, Bruno M

    2015-02-01

    In the last two decades, the third-dimension has become a focus of attention in electron microscopy to better understand the interactions within subcellular compartments. Initially, transmission electron tomography (TEM tomography) was introduced to image the cell volume in semi-thin sections (∼ 500 nm). With the introduction of the focused ion beam scanning electron microscope, a new tool, FIB-SEM tomography, became available to image much larger volumes. During TEM tomography and FIB-SEM tomography, the resin section is exposed to a high electron/ion dose such that the stability of the resin embedded biological sample becomes an important issue. The shrinkage of a resin section in each dimension, especially in depth, is a well-known phenomenon. To ensure the dimensional integrity of the final volume of the cell, it is important to assess the properties of the different resins and determine the formulation which has the best stability in the electron/ion beam. Here, eight different resin formulations were examined. The effects of radiation damage were evaluated after different times of TEM irradiation. To get additional information on mass-loss and the physical properties of the resins (stiffness and adhesion), the topography of the irradiated areas was analysed with atomic force microscopy (AFM). Further, the behaviour of the resins was analysed after ion milling of the surface of the sample with different ion currents. In conclusion, two resin formulations, Hard Plus and the mixture of Durcupan/Epon, emerged that were considerably less affected and reasonably stable in the electron/ion beam and thus suitable for the 3-D investigation of biological samples. PMID:25433274

  7. Multiphotonic Confocal Microscopy 3D imaging: Application to mantle sulfides in sub-arc environment (Avacha Volcano, Kamchatka)

    NASA Astrophysics Data System (ADS)

    Antoine, Bénard; Luc-Serge, Doucet; Sabine, Palle; Dmitri A., Ionov

    2010-05-01

    Petrogenetic relations in igneous rocks are usually studied in natural samples using classical optical microscopy and subsequent geochemical data acquisition. Multiphotonic Laser Scanning Confocal Microscopy (MLSCM) can be a powerful tool to section geological materials optically with sub-micrometric resolution and then generate a three-dimensional (3D) reconstruction (ca. 106 μm3 stack). MLSCM is used here to investigate textural relations of Monosulfide Solid Solution (MSS) with silicate phases in fresh spinel harzburgite xenoliths from the andesitic Avacha volcano (Kamchatka, Russia). The xenoliths contain MSS disseminated in olivine and orthopyroxene (opx) neoblasts as well as MSS-rich quenched magmatic opx veins [1]. First, Reflection Mode (RM) was tested on vein sulfides in resin-impregnated thick (120 μm) polished rock sections. Then we used a combination of Differential Interference Contrast (DIC) with a transmitted light detector, two photons-excited fluorescence (2PEF) and Second Harmonic Generation (SHG). Sequential imaging feature of the Leica TCS-SP2 software was applied. The excitation laser used for 2PEF was a COHERENT MIRA 900 with a 76Hz repetition rate and 800nm wavelength. Image stacks were analysed using ImageJ software [2]. The aim of the tests was to try to discriminate sulfides in silicate matrix as a tool for a better assessment of equilibrium conditions between the two phases. Preliminary results show that Fe-Ni rich MSS from vein and host rock have a strong auto-fluorescence in the Near UV-VIS domain (392-715 nm) whereas silicate matrix is only revealed through DIC. SHG is obtained only from dense nanocentrosymmetrical structures such as embedded medium (organic matter like glue and resin). The three images were recorded sequentially enabling efficient discrimination between the different components of the rock slices. RM permits reconstruction of the complete 3D structure of the rock slice. High resolution (ca. 0.2 μm along X-Y axis vs

  8. Segmentation of vascular structures and hematopoietic cells in 3D microscopy images and quantitative analysis

    NASA Astrophysics Data System (ADS)

    Mu, Jian; Yang, Lin; Kamocka, Malgorzata M.; Zollman, Amy L.; Carlesso, Nadia; Chen, Danny Z.

    2015-03-01

    In this paper, we present image processing methods for quantitative study of how the bone marrow microenvironment changes (characterized by altered vascular structure and hematopoietic cell distribution) caused by diseases or various factors. We develop algorithms that automatically segment vascular structures and hematopoietic cells in 3-D microscopy images, perform quantitative analysis of the properties of the segmented vascular structures and cells, and examine how such properties change. In processing images, we apply local thresholding to segment vessels, and add post-processing steps to deal with imaging artifacts. We propose an improved watershed algorithm that relies on both intensity and shape information and can separate multiple overlapping cells better than common watershed methods. We then quantitatively compute various features of the vascular structures and hematopoietic cells, such as the branches and sizes of vessels and the distribution of cells. In analyzing vascular properties, we provide algorithms for pruning fake vessel segments and branches based on vessel skeletons. Our algorithms can segment vascular structures and hematopoietic cells with good quality. We use our methods to quantitatively examine the changes in the bone marrow microenvironment caused by the deletion of Notch pathway. Our quantitative analysis reveals property changes in samples with deleted Notch pathway. Our tool is useful for biologists to quantitatively measure changes in the bone marrow microenvironment, for developing possible therapeutic strategies to help the bone marrow microenvironment recovery.

  9. Readily Accessible Multiplane Microscopy: 3D Tracking the HIV-1 Genome in Living Cells.

    PubMed

    Itano, Michelle S; Bleck, Marina; Johnson, Daniel S; Simon, Sanford M

    2016-02-01

    Human immunodeficiency virus (HIV)-1 infection and the associated disease AIDS are a major cause of human death worldwide with no vaccine or cure available. The trafficking of HIV-1 RNAs from sites of synthesis in the nucleus, through the cytoplasm, to sites of assembly at the plasma membrane are critical steps in HIV-1 viral replication, but are not well characterized. Here we present a broadly accessible microscopy method that captures multiple focal planes simultaneously, which allows us to image the trafficking of HIV-1 genomic RNAs with high precision. This method utilizes a customization of a commercial multichannel emission splitter that enables high-resolution 3D imaging with single-macromolecule sensitivity. We show with high temporal and spatial resolution that HIV-1 genomic RNAs are most mobile in the cytosol, and undergo confined mobility at sites along the nuclear envelope and in the nucleus and nucleolus. These provide important insights regarding the mechanism by which the HIV-1 RNA genome is transported to the sites of assembly of nascent virions. PMID:26567131

  10. Clean localization super-resolution microscopy for 3D biological imaging

    NASA Astrophysics Data System (ADS)

    Mondal, Partha P.; Curthoys, Nikki M.; Hess, Samuel T.

    2016-01-01

    We propose clean localization microscopy (a variant of fPALM) using a molecule filtering technique. Localization imaging involves acquiring a large number of images containing single molecule signatures followed by one-to-one mapping to render a super-resolution image. In principle, this process can be repeated for other z-planes to construct a 3D image. But, single molecules observed from off-focal planes result in false representation of their presence in the focal plane, resulting in incorrect quantification and analysis. We overcome this with a single molecule filtering technique that imposes constraints on the diffraction limited spot size of single molecules in the image plane. Calibration with sub-diffraction size beads puts a natural cutoff on the actual diffraction-limited size of single molecules in the focal plane. This helps in distinguishing beads present in the focal plane from those in the off-focal planes thereby providing an estimate of the single molecules in the focal plane. We study the distribution of actin (labeled with a photoactivatable CAGE 552 dye) in NIH 3T3 mouse fibroblast cells.

  11. Readily Accessible Multiplane Microscopy: 3D Tracking the HIV-1 Genome in Living Cells.

    PubMed

    Itano, Michelle S; Bleck, Marina; Johnson, Daniel S; Simon, Sanford M

    2016-02-01

    Human immunodeficiency virus (HIV)-1 infection and the associated disease AIDS are a major cause of human death worldwide with no vaccine or cure available. The trafficking of HIV-1 RNAs from sites of synthesis in the nucleus, through the cytoplasm, to sites of assembly at the plasma membrane are critical steps in HIV-1 viral replication, but are not well characterized. Here we present a broadly accessible microscopy method that captures multiple focal planes simultaneously, which allows us to image the trafficking of HIV-1 genomic RNAs with high precision. This method utilizes a customization of a commercial multichannel emission splitter that enables high-resolution 3D imaging with single-macromolecule sensitivity. We show with high temporal and spatial resolution that HIV-1 genomic RNAs are most mobile in the cytosol, and undergo confined mobility at sites along the nuclear envelope and in the nucleus and nucleolus. These provide important insights regarding the mechanism by which the HIV-1 RNA genome is transported to the sites of assembly of nascent virions.

  12. Jamming of a soft granular system of hollow elastic shells in 3D using confocal microscopy

    NASA Astrophysics Data System (ADS)

    Jose, Jissy; van Blaaderen, Alfons; Imhof, Arnout

    2014-03-01

    We introduce a new system for jammed matter research consisting of monodisperse, fluorescent, hollow deformable shells, dispersed in an index matched solvent. The interesting fact about these elastic shells is that they undergo buckling: in each contact one of the shells receives an indentation from its neighbor under compressive stress. This kind of deformation is different from the soft granular systems experimentally studied so far like photo elastic disks, emulsions and foams, where the particles are flattened in the region of contact and conserve their volume. Using confocal microscopy and image analysis routines (ImageJ software) we identified the 3D position of the particles with sub pixel resolution. The force law to find the contact forces between pairs of particle is derived from the theory of elasticity of thin shells, where force is proportional to the square root of indentation depth. The distribution of normalized contact forces showed a similar trend like other jammed systems with a peak around the mean and a tail that decayed faster than exponential away from jamming threshold. Further, we also investigated the structure of the jammed packings and contact number distribution with distance to jamming.

  13. Single particle cryo-electron microscopy and 3-D reconstruction of viruses.

    PubMed

    Guo, Fei; Jiang, Wen

    2014-01-01

    With fast progresses in instrumentation, image processing algorithms, and computational resources, single particle electron cryo-microscopy (cryo-EM) 3-D reconstruction of icosahedral viruses has now reached near-atomic resolutions (3-4 Å). With comparable resolutions and more predictable outcomes, cryo-EM is now considered a preferred method over X-ray crystallography for determination of atomic structure of icosahedral viruses. At near-atomic resolutions, all-atom models or backbone models can be reliably built that allow residue level understanding of viral assembly and conformational changes among different stages of viral life cycle. With the developments of asymmetric reconstruction, it is now possible to visualize the complete structure of a complex virus with not only its icosahedral shell but also its multiple non-icosahedral structural features. In this chapter, we will describe single particle cryo-EM experimental and computational procedures for both near-atomic resolution reconstruction of icosahedral viruses and asymmetric reconstruction of viruses with both icosahedral and non-icosahedral structure components. Procedures for rigorous validation of the reconstructions and resolution evaluations using truly independent de novo initial models and refinements are also introduced.

  14. A resource from 3D electron microscopy of hippocampal neuropil for user training and tool development

    PubMed Central

    Harris, Kristen M.; Spacek, Josef; Bell, Maria Elizabeth; Parker, Patrick H.; Lindsey, Laurence F.; Baden, Alexander D.; Vogelstein, Joshua T.; Burns, Randal

    2015-01-01

    Resurgent interest in synaptic circuitry and plasticity has emphasized the importance of 3D reconstruction from serial section electron microscopy (3DEM). Three volumes of hippocampal CA1 neuropil from adult rat were imaged at X-Y resolution of ~2 nm on serial sections of ~50–60 nm thickness. These are the first densely reconstructed hippocampal volumes. All axons, dendrites, glia, and synapses were reconstructed in a cube (~10 μm3) surrounding a large dendritic spine, a cylinder (~43 μm3) surrounding an oblique dendritic segment (3.4 μm long), and a parallelepiped (~178 μm3) surrounding an apical dendritic segment (4.9 μm long). The data provide standards for identifying ultrastructural objects in 3DEM, realistic reconstructions for modeling biophysical properties of synaptic transmission, and a test bed for enhancing reconstruction tools. Representative synapses are quantified from varying section planes, and microtubules, polyribosomes, smooth endoplasmic reticulum, and endosomes are identified and reconstructed in a subset of dendrites. The original images, traces, and Reconstruct software and files are freely available and visualized at the Open Connectome Project (Data Citation 1). PMID:26347348

  15. Single Particle Cryo-electron Microscopy and 3-D Reconstruction of Viruses

    PubMed Central

    Guo, Fei; Jiang, Wen

    2014-01-01

    With fast progresses in instrumentation, image processing algorithms, and computational resources, single particle electron cryo-microscopy (cryo-EM) 3-D reconstruction of icosahedral viruses has now reached near-atomic resolutions (3–4 Å). With comparable resolutions and more predictable outcomes, cryo-EM is now considered a preferred method over X-ray crystallography for determination of atomic structure of icosahedral viruses. At near-atomic resolutions, all-atom models or backbone models can be reliably built that allow residue level understanding of viral assembly and conformational changes among different stages of viral life cycle. With the developments of asymmetric reconstruction, it is now possible to visualize the complete structure of a complex virus with not only its icosahedral shell but also its multiple non-icosahedral structural features. In this chapter, we will describe single particle cryo-EM experimental and computational procedures for both near-atomic resolution reconstruction of icosahedral viruses and asymmetric reconstruction of viruses with both icosahedral and non-icosahedral structure components. Procedures for rigorous validation of the reconstructions and resolution evaluations using truly independent de novo initial models and refinements are also introduced. PMID:24357374

  16. Photon efficient double-helix PSF microscopy with application to 3D photo-activation localization imaging

    PubMed Central

    Grover, Ginni; Quirin, Sean; Fiedler, Callie; Piestun, Rafael

    2011-01-01

    We present a double-helix point spread function (DH-PSF) based three-dimensional (3D) microscope with efficient photon collection using a phase mask fabricated by gray-level lithography. The system using the phase mask more than doubles the efficiency of current liquid crystal spatial light modulator implementations. We demonstrate the phase mask DH-PSF microscope for 3D photo-activation localization microscopy (PM-DH-PALM) over an extended axial range. PMID:22076263

  17. Neuronal nuclei localization in 3D using level set and watershed segmentation from laser scanning microscopy images

    NASA Astrophysics Data System (ADS)

    Zhu, Yingxuan; Olson, Eric; Subramanian, Arun; Feiglin, David; Varshney, Pramod K.; Krol, Andrzej

    2008-03-01

    Abnormalities of the number and location of cells are hallmarks of both developmental and degenerative neurological diseases. However, standard stereological methods are impractical for assigning each cell's nucleus position within a large volume of brain tissue. We propose an automated approach for segmentation and localization of the brain cell nuclei in laser scanning microscopy (LSM) embryonic mouse brain images. The nuclei in these images are first segmented by using the level set (LS) and watershed methods in each optical plane. The segmentation results are further refined by application of information from adjacent optical planes and prior knowledge of nuclear shape. Segmentation is then followed with an algorithm for 3D localization of the centroid of nucleus (CN). Each volume of tissue is thus represented by a collection of centroids leading to an approximate 10,000-fold reduction in the data set size, as compared to the original image series. Our method has been tested on LSM images obtained from an embryonic mouse brain, and compared to the segmentation and CN localization performed by an expert. The average Euclidian distance between locations of CNs obtained using our method and those obtained by an expert is 1.58+/-1.24 µm, a value well within the ~5 µm average radius of each nucleus. We conclude that our approach accurately segments and localizes CNs within cell dense embryonic tissue.

  18. Image reconstruction for 3D light microscopy with a regularized linear method incorporating a smoothness prior

    NASA Astrophysics Data System (ADS)

    Preza, Chrysanthe; Miller, Michael I.; Conchello, Jose-Angel

    1993-07-01

    We have shown that the linear least-squares (LLS) estimate of the intensities of a 3-D object obtained from a set of optical sections is unstable due to the inversion of small and zero-valued eigenvalues of the point-spread function (PSF) operator. The LLS solution was regularized by constraining it to lie in a subspace spanned by the eigenvectors corresponding to a selected number of the largest eigenvalues. In this paper we extend the regularized LLS solution to a maximum a posteriori (MAP) solution induced by a prior formed from a 'Good's like' smoothness penalty. This approach also yields a regularized linear estimator which reduces noise as well as edge artifacts in the reconstruction. The advantage of the linear MAP (LMAP) estimate over the current regularized LLS (RLLS) is its ability to regularize the inverse problem by smoothly penalizing components in the image associated with small eigenvalues. Computer simulations were performed using a theoretical PSF and a simple phantom to compare the two regularization techniques. It is shown that the reconstructions using the smoothness prior, give superior variance and bias results compared to the RLLS reconstructions. Encouraging reconstructions obtained with the LMAP method from real microscopical images of a 10 micrometers fluorescent bead, and a four-cell Volvox embryo are shown.

  19. Oscillating optical tweezer-based 3-D confocal microrheometer for investigating the intracellular micromechanics and structures

    NASA Astrophysics Data System (ADS)

    Ou-Yang, H. D.; Rickter, E. A.; Pu, C.; Latinovic, O.; Kumar, A.; Mengistu, M.; Lowe-Krentz, L.; Chien, S.

    2005-08-01

    Mechanical properties of living biological cells are important for cells to maintain their shapes, support mechanical stresses and move through tissue matrix. The use of optical tweezers to measure micromechanical properties of cells has recently made significant progresses. This paper presents a new approach, the oscillating optical tweezer cytorheometer (OOTC), which takes advantage of the coherent detection of harmonically modulated particle motions by a lock-in amplifier to increase sensitivity, temporal resolution and simplicity. We demonstrate that OOTC can measure the dynamic mechanical modulus in the frequency range of 0.1-6,000 Hz at a rate as fast as 1 data point per second with submicron spatial resolution. More importantly, OOTC is capable of distinguishing the intrinsic non-random temporal variations from random fluctuations due to Brownian motion; this capability, not achievable by conventional approaches, is particular useful because living systems are highly dynamic and often exhibit non-thermal, rhythmic behavior in a broad time scale from a fraction of a second to hours or days. Although OOTC is effective in measuring the intracellular micromechanical properties, unless we can visualize the cytoskeleton in situ, the mechanical property data would only be as informative as that of "Blind men and the Elephant". To solve this problem, we take two steps, the first, to use of fluorescent imaging to identify the granular structures trapped by optical tweezers, and second, to integrate OOTC with 3-D confocal microscopy so we can take simultaneous, in situ measurements of the micromechanics and intracellular structure in living cells. In this paper, we discuss examples of applying the oscillating tweezer-based cytorheometer for investigating cultured bovine endothelial cells, the identification of caveolae as some of the granular structures in the cell as well as our approach to integrate optical tweezers with a spinning disk confocal microscope.

  20. Integration of 3D anatomical data obtained by CT imaging and 3D optical scanning for computer aided implant surgery

    PubMed Central

    2011-01-01

    Background A precise placement of dental implants is a crucial step to optimize both prosthetic aspects and functional constraints. In this context, the use of virtual guiding systems has been recognized as a fundamental tool to control the ideal implant position. In particular, complex periodontal surgeries can be performed using preoperative planning based on CT data. The critical point of the procedure relies on the lack of accuracy in transferring CT planning information to surgical field through custom-made stereo-lithographic surgical guides. Methods In this work, a novel methodology is proposed for monitoring loss of accuracy in transferring CT dental information into periodontal surgical field. The methodology is based on integrating 3D data of anatomical (impression and cast) and preoperative (radiographic template) models, obtained by both CT and optical scanning processes. Results A clinical case, relative to a fully edentulous jaw patient, has been used as test case to assess the accuracy of the various steps concurring in manufacturing surgical guides. In particular, a surgical guide has been designed to place implants in the bone structure of the patient. The analysis of the results has allowed the clinician to monitor all the errors, which have been occurring step by step manufacturing the physical templates. Conclusions The use of an optical scanner, which has a higher resolution and accuracy than CT scanning, has demonstrated to be a valid support to control the precision of the various physical models adopted and to point out possible error sources. A case study regarding a fully edentulous patient has confirmed the feasibility of the proposed methodology. PMID:21338504

  1. Visual-servoing optical microscopy

    DOEpatents

    Callahan, Daniel E.; Parvin, Bahram

    2009-06-09

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

  2. Visual-servoing optical microscopy

    DOEpatents

    Callahan, Daniel E.; Parvin, Bahram

    2011-05-24

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

  3. Visual-servoing optical microscopy

    DOEpatents

    Callahan, Daniel E; Parvin, Bahram

    2013-10-01

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

  4. 3D phase stepping optical profilometry using a fiber optic Lloyd's mirror

    NASA Astrophysics Data System (ADS)

    Kosoglu, Gulsen; Yuksel, Heba; Inci, M. Naci

    2016-04-01

    This study defines measurements of three-dimensional rigid-body shapes by using a fiber optic Lloyd's mirror. A fiber optic Lloyd's mirror assembly is basically a technique to create an optical interference pattern using the real light point sources and their images. The generated fringe pattern thanks to this technique is deformed when it is projected on an object's surface. The introduced surface profilometry algorithm depends on a multi-step phase shifting process. The deformed fringe patterns containing information of the object's surface profile are captured by a digital CCD camera. While each frames are captured, required π/2 phase shifts for interference fringe pattern are obtained by mechanically sliding the Lloyd assembly via an ordinary micrometer stage. Some preprocess algorithms are applied to the frames and are processed with an algorithm to accomplish 3D topographies. Finally, the continuous data determines the depth information and the surface topography of the object. The experimental setup is simple and low cost to construct, and is insensitive to the ambient temperature fluctuations and environmental vibrations that cause unwanted effects on the projected fringe pattern. Such a fiber optic Lloyd's system which provides an accurate non-contact measurement without contaminating and harming the object surface has a wide range of applications from laser interference based lithography in nano-scale to macro-scale interferometers.

  5. Examination of heterogeneous crossing sequences between toner and rollerball pen strokes by digital microscopy and 3-D laser profilometry.

    PubMed

    Montani, Isabelle; Mazzella, Williams; Guichard, Marion; Marquis, Raymond

    2012-07-01

    The determination of line crossing sequences between rollerball pens and laser printers presents difficulties that may not be overcome using traditional techniques. This research aimed to study the potential of digital microscopy and 3-D laser profilometry to determine line crossing sequences between a toner and an aqueous ink line. Different paper types, rollerball pens, and writing pressure were tested. Correct opinions of the sequence were given for all case scenarios, using both techniques. When the toner was printed before the ink, a light reflection was observed in all crossing specimens, while this was never observed in the other sequence types. The 3-D laser profilometry, more time-consuming, presented the main advantage of providing quantitative results. The findings confirm the potential of the 3-D laser profilometry and demonstrate the efficiency of digital microscopy as a new technique for determining the sequence of line crossings involving rollerball pen ink and toner. PMID:22390180

  6. Inspection, 3D modelling, and rapid prototyping of cultural heritage by means of a 3D optical digitiser

    NASA Astrophysics Data System (ADS)

    Docchio, F.; Sansoni, G.; Trebeschi, M.

    2005-06-01

    This paper presents the activity carried out to perform the three-dimensional acquisition of the "Vittoria Alata", a 2m-high, bronze statue, symbol of our City, located at the Civici Musei di Arte e Storia (S. Giulia) of Brescia. The acquisition of the statue has been performed by using a three-dimensional vision system based on active triangulation and on the projection of non-coherent light. This system, called OPL-3D, represents one of the research products of our Laboratory, which has been active for years in the development of techniques and systems for the contactless acquisition of free-form, complex shapes. The study, originally motivated by the need to explore a new hypothesis on the origin of the "Vittoria Alata", led to its complete digitization and description in terms of both polygonal and NURBS-based models. A suite of copies of the whole statue has been obtained in the framework of the collaboration between the City Museum and the EOS Electro Optical Systems GmbH, located in Munich, Germany. As a first step, one 30 cm-high replica of the whole statue has been produced using a low-resolution triangle model of the statue (3.5 millions of triangles). As a second step, two 1:1 scale copies of the statue have been produced. For them, the Laboratory has provided the high resolution STL file (16 millions of triangles). The paper discusses in detail the hardware and the software facilities used to implement the whole process, and gives a comprehensive description of the results.

  7. Nondestructive optical testing of 3D disperse systems with micro- and nano-particles

    NASA Astrophysics Data System (ADS)

    Bezrukova, Alexandra G.

    2005-04-01

    Nondestructive testing and analysis of three-dimensional (3D) disperse systems (DS) with micro- and nano-particles of different nature by complex of optical compatible methods can provide further progress in on-line control of water and air. The simultaneous analysis of 3D-DS by refractometry, absorbency, fluorescence and by different types of light scattering can help to elaborate the sensing elements for specific impurity control. In our research we have investigated by complex of optical methods different 3D-DS such as: proteins, nucleoproteids, lipoproteids, liposomes, viruses, virosomes, lipid emulsions, blood substitutes, latexes, liquid crystals, biological cells with various form and size (including bacterial cells), metallic powders, clays, kimberlites, zeolites, oils, crude oils, etc., and mixtures -- proteins with nucleic acids, liposomes and viruses, liquid crystals with surfactants, mixtures of clay with bacterial cells, samples of natural and water-supply waters, etc. This experience suggests that the set of optical parameters of so called second class is unique for each 3D-DS. In another words each DS can be characterized by n-dimensional vector in n-dimensional space of optical parameters. Mixtures can be considered as polycomponent and polymodal 3D-DS (such as natural water and air). Due to the fusion of various optical data it is possible to indicate by information statistical theory the inverse physical problem on the presence of impurities in mixtures (viruses, bacteria, oil, metallic particles, etc.), and in this case polymodality of particle size distribution is not an obstacle. Bank of optical data for 3D-DS is the base for analysis by information-statistical method.

  8. Complementary cellophane optic gate and its use for a 3D iPad without glasses

    NASA Astrophysics Data System (ADS)

    Iizuka, K.

    2012-04-01

    A complementary cellophane optic gate was fabricated using a birefringent cellophane sheet. Previous versions of the optic gate required the retardance of the cellophane to be as close to 180° as possible throughout the entire visible wavelength range, which meant it was often difficult to find a cellophane sheet with the right thickness and dispersion characteristics to meet this requirement. The complementary optic gate reported in this paper has no restriction on the thickness, composition, or wavelength range of the cellophane sheet except that the cellophane must have some birefringence. Even with an arbitrary retardance, an extinction ratio of 5 × 10-3 was achieved at λ = 0.63 μm. The optic gate was used to convert an iPad into a 3D display without the need for the observer to wear glasses. The high extinction ratio of the optic gate resulted in a 3D display of supreme quality.

  9. 3D printing of tissue-simulating phantoms as a traceable standard for biomedical optical measurement

    NASA Astrophysics Data System (ADS)

    Dong, Erbao; Wang, Minjie; Shen, Shuwei; Han, Yilin; Wu, Qiang; Xu, Ronald

    2016-01-01

    Optical phantoms are commonly used to validate and calibrate biomedical optical devices in order to ensure accurate measurement of optical properties in biological tissue. However, commonly used optical phantoms are based on homogenous materials that reflect neither optical properties nor multi-layer heterogeneities of biological tissue. Using these phantoms for optical calibration may result in significant bias in biological measurement. We propose to characterize and fabricate tissue simulating phantoms that simulate not only the multi-layer heterogeneities but also optical properties of biological tissue. The tissue characterization module detects tissue structural and functional properties in vivo. The phantom printing module generates 3D tissue structures at different scales by layer-by-layer deposition of phantom materials with different optical properties. The ultimate goal is to fabricate multi-layer tissue simulating phantoms as a traceable standard for optimal calibration of biomedical optical spectral devices.

  10. Air-structured optical fibre drawn from a 3D-printed preform

    NASA Astrophysics Data System (ADS)

    Cook, Kevin; Leon-Saval, Sergio; Canning, John; Reid, Zane; Hossain, Md. Arafat; Peng, Gang-Ding

    2015-09-01

    We report the first optical fibre drawn from a 3D-printed preform. An air-structured polymer preform is printed using a modified butadiene plastic called Bendlay as opposed to the more-common Acrylonitrile Butadiene Styrene (ABS). The preform is subsequently drawn to fibre form at a relatively low temperature of 160 °C and maintains its air-structured cladding holes. Such ability to freely-design and 3D-print complex preform structures, such as photonic bandgap and photonic crystal structures, opens up an exciting new front in optical fibre fabrication.

  11. Near-infrared optical imaging of human brain based on the semi-3D reconstruction algorithm

    NASA Astrophysics Data System (ADS)

    Liu, Ming; Meng, Wei; Qin, Zhuanping; Zhou, Xiaoqing; Zhao, Huijuan; Gao, Feng

    2013-03-01

    In the non-invasive brain imaging with near-infrared light, precise head model is of great significance to the forward model and the image reconstruction. To deal with the individual difference of human head tissues and the problem of the irregular curvature, in this paper, we extracted head structure with Mimics software from the MRI image of a volunteer. This scheme makes it possible to assign the optical parameters to every layer of the head tissues reasonably and solve the diffusion equation with the finite-element analysis. During the solution of the inverse problem, a semi-3D reconstruction algorithm is adopted to trade off the computation cost and accuracy between the full 3-D and the 2-D reconstructions. In this scheme, the changes in the optical properties of the inclusions are assumed either axially invariable or confined to the imaging plane, while the 3-D nature of the photon migration is still retained. This therefore leads to a 2-D inverse issue with the matched 3-D forward model. Simulation results show that comparing to the 3-D reconstruction algorithm, the Semi-3D reconstruction algorithm cut 27% the calculation time consumption.

  12. Quantifying axis orientation in 3D using polarization-sensitive optical coherence tomography (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Liu, Chao; Black, Adam J.; Wang, Hui; Akkin, Taner

    2016-03-01

    The optic axis of birefringent tissues indicates the direction of structural anisotropy. Polarization-sensitive Optical Coherence Tomography (PS-OCT) can provide reflectivity contrast as well as retardance and optic axis orientation contrasts that originate from tissue birefringence. We introduce imaging 3D tissue anisotropy by using a single-camera and polarization-maintaining fiber (PMF) based PS-OCT, which utilizes normal and angled illuminations. Because environmental factors such as the movement of PMF and temperature fluctuations induce arbitrary phase changes, the optic axis orientation measurement has a time-varying offset. In order to measure the absolute axis orientation, we add a calibration path which dynamically provides the arbitrary offset to be subtracted from the relative axis orientation values. The axis orientation on the normal plane is the 2D projection of the fiber direction in 3D space. We propose to characterize the axis orientation in different planes (xy, xy' and x'y planes) by using normal and angled illuminations. This allows calculation of the polar angle that completes the orientation information in 3D. Polarization-based optical systems relying on one illumination angle measure the "apparent birefringence" that light encounters rather than the "true birefringence". Birefringence as a measure of anisotropy is quantified with the orientation information in 3D. The method and validation with a biological tissue are presented. The study can facilitate imaging and mapping the structural connections in anisotropic tissues including the brain.

  13. Study of 3D printing method for GRIN micro-optics devices

    NASA Astrophysics Data System (ADS)

    Wang, P. J.; Yeh, J. A.; Hsu, W. Y.; Cheng, Y. C.; Lee, W.; Wu, N. H.; Wu, C. Y.

    2016-03-01

    Conventional optical elements are based on either refractive or reflective optics theory to fulfill the design specifications via optics performance data. In refractive optical lenses, the refractive index of materials and radius of curvature of element surfaces determine the optical power and wavefront aberrations so that optical performance can be further optimized iteratively. Although gradient index (GRIN) phenomenon in optical materials is well studied for more than a half century, the optics theory in lens design via GRIN materials is still yet to be comprehensively investigated before realistic GRIN lenses are manufactured. In this paper, 3D printing method for manufacture of micro-optics devices with special features has been studied based on methods reported in the literatures. Due to the additive nature of the method, GRIN lenses in micro-optics devices seem to be readily achievable if a design methodology is available. First, derivation of ray-tracing formulae is introduced for all possible structures in GRIN lenses. Optics simulation program is employed for characterization of GRIN lenses with performance data given by aberration coefficients in Zernike polynomial. Finally, a proposed structure of 3D printing machine is described with conceptual illustration.

  14. Viewpoint-independent 3D object segmentation for randomly stacked objects using optical object detection

    NASA Astrophysics Data System (ADS)

    Chen, Liang-Chia; Nguyen, Thanh-Hung; Lin, Shyh-Tsong

    2015-10-01

    This work proposes a novel approach to segmenting randomly stacked objects in unstructured 3D point clouds, which are acquired by a random-speckle 3D imaging system for the purpose of automated object detection and reconstruction. An innovative algorithm is proposed; it is based on a novel concept of 3D watershed segmentation and the strategies for resolving over-segmentation and under-segmentation problems. Acquired 3D point clouds are first transformed into a corresponding orthogonally projected depth map along the optical imaging axis of the 3D sensor. A 3D watershed algorithm based on the process of distance transformation is then performed to detect the boundary, called the edge dam, between stacked objects and thereby to segment point clouds individually belonging to two stacked objects. Most importantly, an object-matching algorithm is developed to solve the over- and under-segmentation problems that may arise during the watershed segmentation. The feasibility and effectiveness of the method are confirmed experimentally. The results reveal that the proposed method is a fast and effective scheme for the detection and reconstruction of a 3D object in a random stack of such objects. In the experiments, the precision of the segmentation exceeds 95% and the recall exceeds 80%.

  15. 3-D Raman Imagery and Atomic Force Microscopy of Ancient Microscopic Fossils

    NASA Astrophysics Data System (ADS)

    Schopf, J.

    2003-12-01

    Investigations of the Precambrian (~540- to ~3,500-Ma-old) fossil record depend critically on identification of authentic microbial fossils. Combined with standard paleontologic studies (e.g., of paleoecologic setting, population structure, cellular morphology, preservational variants), two techniques recently introduced to such studies -- Raman imagery and atomic force microscopy -- can help meet this need. Laser-Raman imagery is a non-intrusive, non-destructive technique that can be used to demonstrate a micron-scale one-to-one correlation between optically discernable morphology and the organic (kerogenous) composition of individual microbial fossils(1,2), a prime indicator of biogencity. Such analyses can be used to characterize the molecular-structural makeup of organic-walled microscopic fossils both in acid-resistant residues and in petrographic thin sections, and whether the fossils analyzed are exposed at the upper surface of, or are embedded within (to depths >65 microns), the section studied. By providing means to map chemically, in three dimensions, whole fossils or parts of such fossils(3), Raman imagery can also show the presence of cell lumina, interior cellular cavities, another prime indicator of biogenicity. Atomic force microscopy (AFM) has been used to visualize the nanometer-scale structure of the kerogenous components of single Precambrian microscopic fossils(4). Capable of analyzing minute fragments of ancient organic matter exposed at the upper surface of thin sections (or of kerogen particles deposited on flat surfaces), such analyses hold promise not only for discriminating between biotic and abiotic micro-objects but for elucidation of the domain size -- and, thus, the degree of graphitization -- of the graphene subunits of the carbonaceous matter analyzed. These techniques -- both new to paleobiology -- can provide useful insight into the biogenicity and geochemical maturity of ancient organic matter. References: (1) Kudryavtsev, A.B. et

  16. 3D reconstruction of coronary arteries using frequency domain optical coherence tomography images and biplane angiography.

    PubMed

    Athanasiou, L S; Bourantas, C V; Siogkas, P K; Sakellarios, A I; Exarchos, T P; Naka, K K; Papafaklis, M I; Michalis, L K; Prati, F; Fotiadis, D I

    2012-01-01

    The aim of this study is to describe a new method for three-dimensional (3D) reconstruction of coronary arteries using Frequency Domain Optical Coherence Tomography (FD-OCT) images. The rationale is to fuse the information about the curvature of the artery, derived from biplane angiographies, with the information regarding the lumen wall, which is produced from the FD-OCT examination. The method is based on a three step approach. In the first step the lumen borders in FD-OCT images are detected. In the second step a 3D curve is produced using the center line of the vessel from the two biplane projections. Finally in the third step the detected lumen borders are placed perpendicularly onto the path based on the centroid of each lumen border. The result is a 3D reconstructed artery produced by all the lumen borders of the FD-OCT pullback representing the 3D arterial geometry of the vessel.

  17. 3D mapping of elastic modulus using shear wave optical micro-elastography

    PubMed Central

    Zhu, Jiang; Qi, Li; Miao, Yusi; Ma, Teng; Dai, Cuixia; Qu, Yueqiao; He, Youmin; Gao, Yiwei; Zhou, Qifa; Chen, Zhongping

    2016-01-01

    Elastography provides a powerful tool for histopathological identification and clinical diagnosis based on information from tissue stiffness. Benefiting from high resolution, three-dimensional (3D), and noninvasive optical coherence tomography (OCT), optical micro-elastography has the ability to determine elastic properties with a resolution of ~10 μm in a 3D specimen. The shear wave velocity measurement can be used to quantify the elastic modulus. However, in current methods, shear waves are measured near the surface with an interference of surface waves. In this study, we developed acoustic radiation force (ARF) orthogonal excitation optical coherence elastography (ARFOE-OCE) to visualize shear waves in 3D. This method uses acoustic force perpendicular to the OCT beam to excite shear waves in internal specimens and uses Doppler variance method to visualize shear wave propagation in 3D. The measured propagation of shear waves agrees well with the simulation results obtained from finite element analysis (FEA). Orthogonal acoustic excitation allows this method to measure the shear modulus in a deeper specimen which extends the elasticity measurement range beyond the OCT imaging depth. The results show that the ARFOE-OCE system has the ability to noninvasively determine the 3D elastic map. PMID:27762276

  18. Solving tolerancing and 3D beam shaping problems by multifunctional wave optical design

    NASA Astrophysics Data System (ADS)

    Buehling, Sven; Wyrowski, Frank

    2000-10-01

    A strategy for designing optical systems that are optimized for multiple optical functions on the basis of wave optics is presented. Each optical function is composed of an input field, a set of fixed system parameters, and a merit function. A design algorithm is proposed which is applicable for optical systems consisting of an transmission operator followed by an arbitrary linear operator. The goal is to find the transmission operator which is optimal for all optical functions simultaneously. In later design steps, the found transmission operator can be transformed to real optical elements, for instance by using the thin element approximation. It is shown that the algorithm is efficiently applicable by investigating two sample applications for multifunctional wave optical design: the design of tolerant systems and 3D beam shaping.

  19. Gabor domain optical coherence microscopy

    NASA Astrophysics Data System (ADS)

    Murali, Supraja

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

  20. Bone tissue phantoms for optical flowmeters at large interoptode spacing generated by 3D-stereolithography

    PubMed Central

    Binzoni, Tiziano; Torricelli, Alessandro; Giust, Remo; Sanguinetti, Bruno; Bernhard, Paul; Spinelli, Lorenzo

    2014-01-01

    A bone tissue phantom prototype allowing to test, in general, optical flowmeters at large interoptode spacings, such as laser-Doppler flowmetry or diffuse correlation spectroscopy, has been developed by 3D-stereolithography technique. It has been demonstrated that complex tissue vascular systems of any geometrical shape can be conceived. Absorption coefficient, reduced scattering coefficient and refractive index of the optical phantom have been measured to ensure that the optical parameters reasonably reproduce real human bone tissue in vivo. An experimental demonstration of a possible use of the optical phantom, utilizing a laser-Doppler flowmeter, is also presented. PMID:25136496

  1. Bone tissue phantoms for optical flowmeters at large interoptode spacing generated by 3D-stereolithography.

    PubMed

    Binzoni, Tiziano; Torricelli, Alessandro; Giust, Remo; Sanguinetti, Bruno; Bernhard, Paul; Spinelli, Lorenzo

    2014-08-01

    A bone tissue phantom prototype allowing to test, in general, optical flowmeters at large interoptode spacings, such as laser-Doppler flowmetry or diffuse correlation spectroscopy, has been developed by 3D-stereolithography technique. It has been demonstrated that complex tissue vascular systems of any geometrical shape can be conceived. Absorption coefficient, reduced scattering coefficient and refractive index of the optical phantom have been measured to ensure that the optical parameters reasonably reproduce real human bone tissue in vivo. An experimental demonstration of a possible use of the optical phantom, utilizing a laser-Doppler flowmeter, is also presented.

  2. Optical design for uniform scanning in MEMS-based 3D imaging lidar.

    PubMed

    Lee, Xiaobao; Wang, Chunhui

    2015-03-20

    This paper proposes a method for the optical system design of uniform scanning in a larger scan field of view (FOV) in 3D imaging lidar. The theoretical formulas are derived for the design scheme. By employing the optical design software ZEMAX, a foldaway uniform scanning optical system based on MEMS has been designed, and the scanning uniformity and spot size of the system on the target plane, perpendicular to optical axis, are analyzed and discussed. Results show that the designed system can scan uniformly within the FOV of 40°×40° with small spot size for the target at distance of about 100 m. PMID:25968504

  3. Bone tissue phantoms for optical flowmeters at large interoptode spacing generated by 3D-stereolithography.

    PubMed

    Binzoni, Tiziano; Torricelli, Alessandro; Giust, Remo; Sanguinetti, Bruno; Bernhard, Paul; Spinelli, Lorenzo

    2014-08-01

    A bone tissue phantom prototype allowing to test, in general, optical flowmeters at large interoptode spacings, such as laser-Doppler flowmetry or diffuse correlation spectroscopy, has been developed by 3D-stereolithography technique. It has been demonstrated that complex tissue vascular systems of any geometrical shape can be conceived. Absorption coefficient, reduced scattering coefficient and refractive index of the optical phantom have been measured to ensure that the optical parameters reasonably reproduce real human bone tissue in vivo. An experimental demonstration of a possible use of the optical phantom, utilizing a laser-Doppler flowmeter, is also presented. PMID:25136496

  4. Rapid 3D µ-printing of polymer optical whispering-gallery mode resonators.

    PubMed

    Wu, Jushuai; Guo, Xin; Zhang, A Ping; Tam, Hwa-Yaw

    2015-11-16

    A novel microfabrication method for rapid printing of polymer optical whispering-gallery mode (WGM) resonators is presented. A 3D micro-printing technology based on high-speed optical spatial modulator (SLM) and high-power UV light source is developed to fabricate suspended-disk WGM resonator array using SU-8 photoresist. The optical spectral responses of the fabricated polymer WGM resonators were measured with a biconically tapered optical fiber. Experimental results reveal that the demonstrated method is very flexible and time-saving for rapid fabrication of complex polymer WGM resonators. PMID:26698452

  5. Tracking left ventricular borders in 3D echocardiographic sequences using motion-guided optical flow

    NASA Astrophysics Data System (ADS)

    Leung, K. Y. Esther; Danilouchkine, Mikhail G.; van Stralen, Marijn; de Jong, Nico; van der Steen, Antonius F. W.; Bosch, Johan G.

    2009-02-01

    For obtaining quantitative and objective functional parameters from three-dimensional (3D) echocardiographic sequences, automated segmentation methods may be preferable to cumbersome manual delineation of 3D borders. In this study, a novel optical-flow based tracking method is proposed for propagating 3D endocardial contours of the left ventricle throughout the cardiac cycle. To take full advantage of the time-continuous nature of cardiac motion, a statistical motion model was explicitly embedded in the optical flow solution. The cardiac motion was modeled as frame-to-frame affine transforms, which were extracted using Procrustes analysis on a set of training contours. Principal component analysis was applied to obtain a compact model of cardiac motion throughout the whole cardiac cycle. The parameters of this model were resolved in an optical flow manner, via spatial and temporal gradients in image intensity. The algorithm was tested on 36 noncontrast and 28 contrast enhanced 3D echocardiographic sequences in a leave-one-out manner. Good results were obtained using a combination of the proposed motion-guided method and a purely data-driven optical flow approach. The improvement was particularly noticeable in areas where the LV wall was obscured by image artifacts. In conclusion, the results show the applicability of the proposed method in clinical quality echocardiograms.

  6. Alterations of filopodia by near infrared photoimmunotherapy: evaluation with 3D low-coherent quantitative phase microscopy

    PubMed Central

    Nakamura, Yuko; Nagaya, Tadanobu; Sato, Kazuhide; Harada, Toshiko; Okuyama, Shuhei; Choyke, Peter L.; Yamauchi, Toyohiko; Kobayashi, Hisataka

    2016-01-01

    Filopodia are highly organized cellular membrane structures that facilitate intercellular communication. Near infrared photoimmunotherapy (NIR-PIT) is a newly developed cancer treatment that causes necrotic cell death. Three-dimensional low-coherent quantitative phase microscopy (3D LC-QPM) is based on a newly established low-coherent interference microscope designed to obtain serial topographic images of the cellular membrane. Herein, we report rapid involution of filopodia after NIR-PIT using 3D LC-QPM. For 3T3/HER2 cells, the number of filopodia decreased immediately after treatment with significant differences. Volume and relative height of 3T3/HER2 cells increased immediately after NIR light exposure, but significant differences were not observed. Thus, disappearance of filopodia, evaluated by 3D LC-QPM, is an early indicator of cell membrane damage after NIR-PIT. PMID:27446702

  7. Wide-field hyperspectral 3D imaging of functionalized gold nanoparticles targeting cancer cells by reflected light microscopy.

    PubMed

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

    2015-05-01

    We present a new hyperspectral reflected light microscopy system with a scanned broadband supercontinuum light source. This wide-field and low phototoxic hyperspectral imaging system has been successful for performing spectral three-dimensional (3D) localization and spectroscopic identification of CD44-targeted PEGylated AuNPs in fixed cell preparations. Such spatial and spectral information is essential for the improvement of nanoplasmonic-based imaging, disease detection and treatment in complex biological environment. The presented system can be used for real-time 3D NP tracking as spectral sensors, thus providing new avenues in the spatio-temporal characterization and detection of bioanalytes. 3D image of the distribution of functionalized AuNPs attached to CD44-expressing MDA-MB-231 human cancer cells. PMID:24961507

  8. Imaging bacterial 3D motion using digital in-line holographic microscopy and correlation-based de-noising algorithm

    PubMed Central

    Molaei, Mehdi; Sheng, Jian

    2014-01-01

    Abstract: Better understanding of bacteria environment interactions in the context of biofilm formation requires accurate 3-dimentional measurements of bacteria motility. Digital Holographic Microscopy (DHM) has demonstrated its capability in resolving 3D distribution and mobility of particulates in a dense suspension. Due to their low scattering efficiency, bacteria are substantially difficult to be imaged by DHM. In this paper, we introduce a novel correlation-based de-noising algorithm to remove the background noise and enhance the quality of the hologram. Implemented in conjunction with DHM, we demonstrate that the method allows DHM to resolve 3-D E. coli bacteria locations of a dense suspension (>107 cells/ml) with submicron resolutions (<0.5 µm) over substantial depth and to obtain thousands of 3D cell trajectories. PMID:25607177

  9. Alterations of filopodia by near infrared photoimmunotherapy: evaluation with 3D low-coherent quantitative phase microscopy.

    PubMed

    Nakamura, Yuko; Nagaya, Tadanobu; Sato, Kazuhide; Harada, Toshiko; Okuyama, Shuhei; Choyke, Peter L; Yamauchi, Toyohiko; Kobayashi, Hisataka

    2016-07-01

    Filopodia are highly organized cellular membrane structures that facilitate intercellular communication. Near infrared photoimmunotherapy (NIR-PIT) is a newly developed cancer treatment that causes necrotic cell death. Three-dimensional low-coherent quantitative phase microscopy (3D LC-QPM) is based on a newly established low-coherent interference microscope designed to obtain serial topographic images of the cellular membrane. Herein, we report rapid involution of filopodia after NIR-PIT using 3D LC-QPM. For 3T3/HER2 cells, the number of filopodia decreased immediately after treatment with significant differences. Volume and relative height of 3T3/HER2 cells increased immediately after NIR light exposure, but significant differences were not observed. Thus, disappearance of filopodia, evaluated by 3D LC-QPM, is an early indicator of cell membrane damage after NIR-PIT. PMID:27446702

  10. Web-based visualisation and analysis of 3D electron-microscopy data from EMDB and PDB☆

    PubMed Central

    Lagerstedt, Ingvar; Moore, William J.; Patwardhan, Ardan; Sanz-García, Eduardo; Best, Christoph; Swedlow, Jason R.; Kleywegt, Gerard J.

    2013-01-01

    The Protein Data Bank in Europe (PDBe) has developed web-based tools for the visualisation and analysis of 3D electron microscopy (3DEM) structures in the Electron Microscopy Data Bank (EMDB) and Protein Data Bank (PDB). The tools include: (1) a volume viewer for 3D visualisation of maps, tomograms and models, (2) a slice viewer for inspecting 2D slices of tomographic reconstructions, and (3) visual analysis pages to facilitate analysis and validation of maps, tomograms and models. These tools were designed to help non-experts and experts alike to get some insight into the content and assess the quality of 3DEM structures in EMDB and PDB without the need to install specialised software or to download large amounts of data from these archives. The technical challenges encountered in developing these tools, as well as the more general considerations when making archived data available to the user community through a web interface, are discussed. PMID:24113529

  11. Web-based visualisation and analysis of 3D electron-microscopy data from EMDB and PDB.

    PubMed

    Lagerstedt, Ingvar; Moore, William J; Patwardhan, Ardan; Sanz-García, Eduardo; Best, Christoph; Swedlow, Jason R; Kleywegt, Gerard J

    2013-11-01

    The Protein Data Bank in Europe (PDBe) has developed web-based tools for the visualisation and analysis of 3D electron microscopy (3DEM) structures in the Electron Microscopy Data Bank (EMDB) and Protein Data Bank (PDB). The tools include: (1) a volume viewer for 3D visualisation of maps, tomograms and models, (2) a slice viewer for inspecting 2D slices of tomographic reconstructions, and (3) visual analysis pages to facilitate analysis and validation of maps, tomograms and models. These tools were designed to help non-experts and experts alike to get some insight into the content and assess the quality of 3DEM structures in EMDB and PDB without the need to install specialised software or to download large amounts of data from these archives. The technical challenges encountered in developing these tools, as well as the more general considerations when making archived data available to the user community through a web interface, are discussed.

  12. Determination of 3D optic axis orientation in cartilage by polarization-sensitive optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Ugryumova, Nadya; Matcher, Stephen J.

    2007-02-01

    Polarization-sensitive optical coherence tomography has been used to solve fast-axis fibre orientation in three dimension space. Previously we have demonstrated that the apparent variations in polar angle orientation of collagen fibers along sagittal ridge of equine third metacarpophalangeal joint exist. A quantitative method based on multiple angles of illumination has been proposed to determine the polar angle of the collagen fibers. This method however ignored the full 3-D structure by assuming that the collagen fibers long-axis lay within the plane of incidence. A new quantitative method based on the theory of light propagation in uniaxial materials is described which avoids this assumption. To test this method we have performed control experiments on a sample of equine tendon (this tissue has well defined c-axis lying along the long-axis of the tendon). Several samples of tendon were cut to achieve a planar surface inclined at -20° to the long axis. Additional 30° rotation provided non-zero azimuthal angle. The surface was then imaged using incident beam angles -40°, -20°, 0, +20°, +40° in two orthogonal planes. Values for both the polar and azimuthal angles were then derived using a numerical optimisation procedure. Results agreed qualitatively with the nominal values but suggested that the accuracy was limited by our method of determining the apparent birefringence.

  13. Determination of 3D optic axis orientation in cartilage by polarization-sensitive optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Ugryumova, Nadya; Bonesi, Marco; Matcher, Stephen J.

    2008-02-01

    Polarization-sensitive optical coherence tomography has been used to solve fast-axis fibre orientation in three dimension space. Previously we have demonstrated that the apparent variations in polar angle orientation of collagen fibers along sagittal ridge of equine third metacarpophalangeal joint exist. A quantitative method based on multiple angles of illumination has been proposed to determine the polar angle of the collagen fibers. This method however ignored the full 3D structure by assuming that the collagen fibers long-axis lay within the plane of incidence. A new quantitative method based on the theory of light propagation in uniaxial materials is described which avoids this assumption. To test this method we have performed control experiments on a sample of equine tendon (this tissue has well defined c-axis lying along the long-axis of the tendon). Several samples of tendon were cut to achieve a planar surface inclined at -20° to the long axis. Additional 30° rotation provided non-zero azimuthal angle. The surface was then imaged using incident beam angles -40°, -20°, 0, +20°, +40° in two orthogonal planes. Values for both the polar and azimuthal angles were then derived using a numerical optimisation procedure. Results agreed qualitatively with the nominal values but suggested that the accuracy was limited by our method of determining the apparent birefringence.

  14. Disposable optics for microscopy diagnostics

    PubMed Central

    Vilmi, Pauliina; Varjo, Sami; Sliz, Rafal; Hannuksela, Jari; Fabritius, Tapio

    2015-01-01

    The point-of-care testing (POCT) is having increasing role on modern health care systems due to a possibility to perform tests for patients conveniently and immediately. POCT includes lot of disposable devices because of the environment they are often used. For a disposable system to be reasonably utilized, it needs to be high in quality but low in price. Optics based POCT systems are interesting approach to be developed, and here we describe a low-cost fabrication process for microlens arrays for microscopy. Lens arrays having average lens diameter of 222 μm with 300 μm lens pitch were fabricated. The lenses were characterized to have standard deviation of 0.06 μm in height and 4.61 μm in diameter. The resolution limit of 3.9μm is demonstrated with real images, and the images were compared with ones made with glass and polycarbonate lens arrays. The image quality is at the same level than with the glass lenses and the manufacturing costs are very low, thus making them suitable for POCT applications. PMID:26586153

  15. Disposable optics for microscopy diagnostics.

    PubMed

    Vilmi, Pauliina; Varjo, Sami; Sliz, Rafal; Hannuksela, Jari; Fabritius, Tapio

    2015-11-20

    The point-of-care testing (POCT) is having increasing role on modern health care systems due to a possibility to perform tests for patients conveniently and immediately. POCT includes lot of disposable devices because of the environment they are often used. For a disposable system to be reasonably utilized, it needs to be high in quality but low in price. Optics based POCT systems are interesting approach to be developed, and here we describe a low-cost fabrication process for microlens arrays for microscopy. Lens arrays having average lens diameter of 222 μm with 300 μm lens pitch were fabricated. The lenses were characterized to have standard deviation of 0.06 μm in height and 4.61 μm in diameter. The resolution limit of 3.9μm is demonstrated with real images, and the images were compared with ones made with glass and polycarbonate lens arrays. The image quality is at the same level than with the glass lenses and the manufacturing costs are very low, thus making them suitable for POCT applications.

  16. Disposable optics for microscopy diagnostics

    NASA Astrophysics Data System (ADS)

    Vilmi, Pauliina; Varjo, Sami; Sliz, Rafal; Hannuksela, Jari; Fabritius, Tapio

    2015-11-01

    The point-of-care testing (POCT) is having increasing role on modern health care systems due to a possibility to perform tests for patients conveniently and immediately. POCT includes lot of disposable devices because of the environment they are often used. For a disposable system to be reasonably utilized, it needs to be high in quality but low in price. Optics based POCT systems are interesting approach to be developed, and here we describe a low-cost fabrication process for microlens arrays for microscopy. Lens arrays having average lens diameter of 222 μm with 300 μm lens pitch were fabricated. The lenses were characterized to have standard deviation of 0.06 μm in height and 4.61 μm in diameter. The resolution limit of 3.9μm is demonstrated with real images, and the images were compared with ones made with glass and polycarbonate lens arrays. The image quality is at the same level than with the glass lenses and the manufacturing costs are very low, thus making them suitable for POCT applications.

  17. 3D scanning electron microscopy applied to surface characterization of fluorosed dental enamel.

    PubMed

    Limandri, Silvina; Galván Josa, Víctor; Valentinuzzi, María Cecilia; Chena, María Emilia; Castellano, Gustavo

    2016-05-01

    The enamel surfaces of fluorotic teeth were studied by scanning electron stereomicroscopy. Different whitening treatments were applied to 25 pieces to remove stains caused by fluorosis and their surfaces were characterized by stereomicroscopy in order to obtain functional and amplitude parameters. The topographic features resulting for each treatment were determined through these parameters. The results obtained show that the 3D reconstruction achieved from the SEM stereo pairs is a valuable potential alternative for the surface characterization of this kind of samples.

  18. Simple 3D images from fossil and recent micromaterial using light microscopy.

    PubMed

    Haug, J T; Haug, C; Maas, A; Fayers, S R; Trewin, N H; Waloszek, D

    2009-01-01

    Abstract We present a technique for extracting 3D information from small-scale fossil and Recent material and give a summary of other contemporary techniques for 3D methods of investigation. The only hardware needed for the here-presented technique is a microscope that can perform dark field and/or differential interference contrast with a mounted digital camera and a computer. Serial images are taken while the focus is successively shifted from the uppermost end of the specimen to the lowermost end, resulting in about 200 photographs. The data are then processed almost completely automatically by successive use of three freely available programs. Firstly, the stack of images is aligned by the use of CombineZM, which is used to produce a combined image with a high depth of field. Secondly, the aligned images are cropped and sharp edges extracted with the aid of ImageJ. Thirdly, although ImageJ is also capable of producing 3D representations, we preferred to process the image stack further using osirix as it has the facility to export various formats. One of the interesting export formats is a virtual Quicktime movie file (QTVR), which can be used for documentation, and stereo images can also be produced from this Quicktime VR. This method is easy to apply and can be used for documenting specimens in 3D (at least some aspects) without having to prepare them. Therefore, it is particularly useful as a safe method for documenting limited material, before using methods that may destroy the specimen of interest, or to investigate type material that cannot be treated with any preparatory technique. As light microscopes are available in most labs and free computer programs are easily accessible, this method can be readily applied. PMID:19196416

  19. A 3D integral imaging optical see-through head-mounted display.

    PubMed

    Hua, Hong; Javidi, Bahram

    2014-06-01

    An optical see-through head-mounted display (OST-HMD), which enables optical superposition of digital information onto the direct view of the physical world and maintains see-through vision to the real world, is a vital component in an augmented reality (AR) system. A key limitation of the state-of-the-art OST-HMD technology is the well-known accommodation-convergence mismatch problem caused by the fact that the image source in most of the existing AR displays is a 2D flat surface located at a fixed distance from the eye. In this paper, we present an innovative approach to OST-HMD designs by combining the recent advancement of freeform optical technology and microscopic integral imaging (micro-InI) method. A micro-InI unit creates a 3D image source for HMD viewing optics, instead of a typical 2D display surface, by reconstructing a miniature 3D scene from a large number of perspective images of the scene. By taking advantage of the emerging freeform optical technology, our approach will result in compact, lightweight, goggle-style AR display that is potentially less vulnerable to the accommodation-convergence discrepancy problem and visual fatigue. A proof-of-concept prototype system is demonstrated, which offers a goggle-like compact form factor, non-obstructive see-through field of view, and true 3D virtual display.

  20. Performance of an improved first generation optical CT scanner for 3D dosimetry

    NASA Astrophysics Data System (ADS)

    Qian, Xin; Adamovics, John; Wuu, Cheng-Shie

    2013-12-01

    Performance analysis of a modified 3D dosimetry optical scanner based on the first generation optical CT scanner OCTOPUS is presented. The system consists of PRESAGE™ dosimeters, the modified 3D scanner, and a new developed in-house user control panel written in Labview program which provides more flexibility to optimize mechanical control and data acquisition technique. The total scanning time has been significantly reduced from initial 8 h to ∼2 h by using the modified scanner. The functional performance of the modified scanner has been evaluated in terms of the mechanical integrity uncertainty of the data acquisition process. Optical density distribution comparison between the modified scanner, OCTOPUS and the treatment plan system has been studied. It has been demonstrated that the agreement between the modified scanner and treatment plans is comparable with that between the OCTOPUS and treatment plans.

  1. Analytical description of 3D optical pulse diffraction by a phase-shifted Bragg grating.

    PubMed

    Golovastikov, Nikita V; Bykov, Dmitry A; Doskolovich, Leonid L; Soifer, Victor A

    2016-08-22

    Diffraction of a three-dimensional (3D) spatiotemporal optical pulse by a phase-shifted Bragg grating (PSBG) is considered. The pulse diffraction is described in terms of signal transmission through a linear system with a transfer function determined by the reflection or transmission coefficient of the PSBG. Resonant approximations of the reflection and transmission coefficients of the PSBG as functions of the angular frequency and the in-plane component of the wave vector are obtained. Using these approximations, a hyperbolic partial differential equation (Klein-Gordon equation) describing a general class of transformations of the incident 3D pulse envelope is derived. A solution to this equation is found in the form of a convolution integral. The presented rigorous simulation results fully confirm the proposed theoretical description. The obtained results may find application in the design of new devices for spatiotemporal pulse shaping and for optical information processing and analog optical computing. PMID:27557167

  2. Generation of nearly 3D-unpolarized evanescent optical near fields using total internal reflection.

    PubMed

    Hassinen, Timo; Popov, Sergei; Friberg, Ari T; Setälä, Tero

    2016-07-01

    We analyze the time-domain partial polarization of optical fields composed of two evanescent waves created in total internal reflection by random electromagnetic beams with orthogonal planes of incidence. We show that such a two-beam configuration enables to generate nearly unpolarized, genuine three-component (3D) near fields. This result complements earlier studies on spectral polarization, which state that at least three symmetrically propagating beams are required to produce a 3D-unpolarized near field. The degree of polarization of the near field can be controlled by adjusting the polarization states and mutual correlation of the incident beams.

  3. Generation of nearly 3D-unpolarized evanescent optical near fields using total internal reflection.

    PubMed

    Hassinen, Timo; Popov, Sergei; Friberg, Ari T; Setälä, Tero

    2016-07-01

    We analyze the time-domain partial polarization of optical fields composed of two evanescent waves created in total internal reflection by random electromagnetic beams with orthogonal planes of incidence. We show that such a two-beam configuration enables to generate nearly unpolarized, genuine three-component (3D) near fields. This result complements earlier studies on spectral polarization, which state that at least three symmetrically propagating beams are required to produce a 3D-unpolarized near field. The degree of polarization of the near field can be controlled by adjusting the polarization states and mutual correlation of the incident beams. PMID:27367071

  4. Real-time visualization of 3-D dynamic microscopic objects using optical diffraction tomography.

    PubMed

    Kim, Kyoohyun; Kim, Kyung Sang; Park, Hyunjoo; Ye, Jong Chul; Park, Yongkeun

    2013-12-30

    3-D refractive index (RI) distribution is an intrinsic bio-marker for the chemical and structural information about biological cells. Here we develop an optical diffraction tomography technique for the real-time reconstruction of 3-D RI distribution, employing sparse angle illumination and a graphic processing unit (GPU) implementation. The execution time for the tomographic reconstruction is 0.21 s for 96(3) voxels, which is 17 times faster than that of a conventional approach. We demonstrated the real-time visualization capability with imaging the dynamics of Brownian motion of an anisotropic colloidal dimer and the dynamic shape change in a red blood cell upon shear flow.

  5. Optical design of wavelength selective CPVT system with 3D/2D hybrid concentration

    NASA Astrophysics Data System (ADS)

    Ahmad, N.; Ijiro, T.; Yamada, N.; Kawaguchi, T.; Maemura, T.; Ohashi, H.

    2012-10-01

    Optical design of a concentrating photovoltaic/thermal (CPVT) system is carried out. Using wavelength-selective optics, the system demonstrates 3-D concentration onto a solar cell and 2-D concentration onto a thermal receiver. Characteristics of the two types of concentrator systems are examined with ray-tracing analysis. The first system is a glazed mirror-based concentrator system mounted on a 2-axis pedestal tracker. The size of the secondary optical element is minimized to decrease the cost of the system, and it has a wavelength-selective function for performing 3-D concentration onto a solar cell and 2-D concentration onto a thermal receiver. The second system is a non-glazed beamdown concentrator system containing parabolic mirrors in the lower part. The beam-down selective mirror performs 3-D concentration onto a solar cell placed above the beam-down selective mirror, and 2-D concentration down to a thermal receiver placed at the bottom level. The system is mounted on a two-axis carousel tracker. A parametric study is performed for those systems with different geometrical 2-D/3-D concentration ratios. Wavelength-selective optics such as hot/cold mirrors and spectrum-splitting technologies are taken into account in the analysis. Results show reduced heat load on the solar cell and increased total system efficiency compared to a non-selective CPV system. Requirements for the wavelength-selective properties are elucidated. It is also shown that the hybrid concept with 2-D concentration onto a thermal receiver and 3-D concentration onto a solar cell has an advantageous geometry because of the high total system efficiency and compatibility with the piping arrangement of the thermal receiver.

  6. Optical microscopy versus scanning electron microscopy in urolithiasis.

    PubMed

    Marickar, Y M Fazil; Lekshmi, P R; Varma, Luxmi; Koshy, Peter

    2009-10-01

    Stone analysis is incompletely done in many clinical centers. Identification of the stone component is essential for deciding future prophylaxis. X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM) still remains a distant dream for routine hospital work. It is in this context that optical microscopy is suggested as an alternate procedure. The objective of this article was to assess the utility of an optical microscope which gives magnification of up to 40x and gives clear picture of the surface of the stones. In order to authenticate the morphological analysis of urinary stones, SEM and elemental distribution analysis were performed. A total of 250 urinary stones of different compositions were collected from stone clinic, photographed, observed under an optical microscope, and optical photographs were taken at different angles. Twenty-five representative samples among these were gold sputtered to make them conductive and were fed into the SEM machine. Photographs of the samples were taken at different angles at magnifications up to 4,000. Elemental distribution analysis (EDAX) was done to confirm the composition. The observations of the two studies were compared. The different appearances of the stones under optical illuminated microscopy were mostly standardized appearances, namely bosselations of pure whewellite, spiculations of weddellite, bright yellow colored appearance of uric acid, and dirty white amorphous appearance of phosphates. SEM and EDAX gave clearer pictures and gave added confirmation of the stone composition. From the references thus obtained, it was possible to confirm the composition by studying the optical microscopic pictures. Higher magnification capacity of the SEM and the EDAX patterns are useful to give reference support for performing optical microscopy work. After standardization, routine analysis can be performed with optical microscopy. The advantage of the optical microscope is that, it

  7. Introductory review on `Flying Triangulation': a motion-robust optical 3D measurement principle

    NASA Astrophysics Data System (ADS)

    Ettl, Svenja

    2015-04-01

    'Flying Triangulation' (FlyTri) is a recently developed principle which allows for a motion-robust optical 3D measurement of rough surfaces. It combines a simple sensor with sophisticated algorithms: a single-shot sensor acquires 2D camera images. From each camera image, a 3D profile is generated. The series of 3D profiles generated are aligned to one another by algorithms, without relying on any external tracking device. It delivers real-time feedback of the measurement process which enables an all-around measurement of objects. The principle has great potential for small-space acquisition environments, such as the measurement of the interior of a car, and motion-sensitive measurement tasks, such as the intraoral measurement of teeth. This article gives an overview of the basic ideas and applications of FlyTri. The main challenges and their solutions are discussed. Measurement examples are also given to demonstrate the potential of the measurement principle.

  8. 3D Optical Measuring Systems and Laser Technologies for Scientific and Industrial Applications

    NASA Astrophysics Data System (ADS)

    Chugui, Yu.; Verkhoglyad, A.; Poleshchuk, A.; Korolkov, V.; Sysoev, E.; Zavyalov, P.

    2013-12-01

    Modern industry and science require novel 3D optical measuring systems and laser technologies with micro/nanometer resolution for solving actual problems. Such systems, including the 3D dimensional inspection of ceramic parts for electrotechnical industry, laser inspection of wheel pair diagnostic for running trains and 3D superresolution low-coherent micro- /nanoprofilometers are presented. The newest results in the field of laser technologies for high-precision synthesis of microstructures by updated image generator using the semiconductor laser are given. The measuring systems and the laser image generator developed and produced by TDI SIE and IAE SB RAS have been tested by customers and used in different branches of industry and science.

  9. An optical system for detecting 3D high-speed oscillation of a single ultrasound microbubble

    PubMed Central

    Liu, Yuan; Yuan, Baohong

    2013-01-01

    As contrast agents, microbubbles have been playing significant roles in ultrasound imaging. Investigation of microbubble oscillation is crucial for microbubble characterization and detection. Unfortunately, 3-dimensional (3D) observation of microbubble oscillation is challenging and costly because of the bubble size—a few microns in diameter—and the high-speed dynamics under MHz ultrasound pressure waves. In this study, a cost-efficient optical confocal microscopic system combined with a gated and intensified charge-coupled device (ICCD) camera were developed to detect 3D microbubble oscillation. The capability of imaging microbubble high-speed oscillation with much lower costs than with an ultra-fast framing or streak camera system was demonstrated. In addition, microbubble oscillations along both lateral (x and y) and axial (z) directions were demonstrated. Accordingly, this system is an excellent alternative for 3D investigation of microbubble high-speed oscillation, especially when budgets are limited. PMID:24049677

  10. Mesoscopic in vivo 3-D tracking of sparse cell populations using angular multiplexed optical projection tomography

    PubMed Central

    Chen, Lingling; Alexandrov, Yuriy; Kumar, Sunil; Andrews, Natalie; Dallman, Margaret J.; French, Paul M. W.; McGinty, James

    2015-01-01

    We describe an angular multiplexed imaging technique for 3-D in vivo cell tracking of sparse cell distributions and optical projection tomography (OPT) with superior time-lapse resolution and a significantly reduced light dose compared to volumetric time-lapse techniques. We demonstrate that using dual axis OPT, where two images are acquired simultaneously at different projection angles, can enable localization and tracking of features in 3-D with a time resolution equal to the camera frame rate. This is achieved with a 200x reduction in light dose compared to an equivalent volumetric time-lapse single camera OPT acquisition with 200 projection angles. We demonstrate the application of this technique to mapping the 3-D neutrophil migration pattern observed over ~25.5 minutes in a live 2 day post-fertilisation transgenic LysC:GFP zebrafish embryo following a tail wound. PMID:25909009

  11. An optical system for detecting 3D high-speed oscillation of a single ultrasound microbubble.

    PubMed

    Liu, Yuan; Yuan, Baohong

    2013-01-01

    As contrast agents, microbubbles have been playing significant roles in ultrasound imaging. Investigation of microbubble oscillation is crucial for microbubble characterization and detection. Unfortunately, 3-dimensional (3D) observation of microbubble oscillation is challenging and costly because of the bubble size-a few microns in diameter-and the high-speed dynamics under MHz ultrasound pressure waves. In this study, a cost-efficient optical confocal microscopic system combined with a gated and intensified charge-coupled device (ICCD) camera were developed to detect 3D microbubble oscillation. The capability of imaging microbubble high-speed oscillation with much lower costs than with an ultra-fast framing or streak camera system was demonstrated. In addition, microbubble oscillations along both lateral (x and y) and axial (z) directions were demonstrated. Accordingly, this system is an excellent alternative for 3D investigation of microbubble high-speed oscillation, especially when budgets are limited. PMID:24049677

  12. Determination of the positions and orientations of concentrated rod-like colloids from 3D microscopy data.

    PubMed

    Besseling, T H; Hermes, M; Kuijk, A; de Nijs, B; Deng, T-S; Dijkstra, M; Imhof, A; van Blaaderen, A

    2015-05-20

    Confocal microscopy in combination with real-space particle tracking has proven to be a powerful tool in scientific fields such as soft matter physics, materials science and cell biology. However, 3D tracking of anisotropic particles in concentrated phases remains not as optimized compared to algorithms for spherical particles. To address this problem, we developed a new particle-fitting algorithm that can extract the positions and orientations of fluorescent rod-like particles from three dimensional confocal microscopy data stacks. The algorithm is tailored to work even when the fluorescent signals of the particles overlap considerably and a threshold method and subsequent clusters analysis alone do not suffice. We demonstrate that our algorithm correctly identifies all five coordinates of uniaxial particles in both a concentrated disordered phase and a liquid-crystalline smectic-B phase. Apart from confocal microscopy images, we also demonstrate that the algorithm can be used to identify nanorods in 3D electron tomography reconstructions. Lastly, we determined the accuracy of the algorithm using both simulated and experimental confocal microscopy data-stacks of diffusing silica rods in a dilute suspension. This novel particle-fitting algorithm allows for the study of structure and dynamics in both dilute and dense liquid-crystalline phases (such as nematic, smectic and crystalline phases) as well as the study of the glass transition of rod-like particles in three dimensions on the single particle level. PMID:25922931

  13. 3D scanning electron microscopy applied to surface characterization of fluorosed dental enamel.

    PubMed

    Limandri, Silvina; Galván Josa, Víctor; Valentinuzzi, María Cecilia; Chena, María Emilia; Castellano, Gustavo

    2016-05-01

    The enamel surfaces of fluorotic teeth were studied by scanning electron stereomicroscopy. Different whitening treatments were applied to 25 pieces to remove stains caused by fluorosis and their surfaces were characterized by stereomicroscopy in order to obtain functional and amplitude parameters. The topographic features resulting for each treatment were determined through these parameters. The results obtained show that the 3D reconstruction achieved from the SEM stereo pairs is a valuable potential alternative for the surface characterization of this kind of samples. PMID:26930005

  14. Site-Specific Cryo-focused Ion Beam Sample Preparation Guided by 3D Correlative Microscopy.

    PubMed

    Arnold, Jan; Mahamid, Julia; Lucic, Vladan; de Marco, Alex; Fernandez, Jose-Jesus; Laugks, Tim; Mayer, Tobias; Hyman, Anthony A; Baumeister, Wolfgang; Plitzko, Jürgen M

    2016-02-23

    The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assemblies within such limited volumes requires precise targeting of the FIB milling process. In this study, we present the development of a cryo-stage allowing for spinning-disk confocal light microscopy at cryogenic temperatures and describe the incorporation of the new hardware into existing workflows for cellular sample preparation by cryo-FIB. Introduction of fiducial markers and subsequent computation of three-dimensional coordinate transformations provide correlation between light microscopy and scanning electron microscopy/FIB. The correlative approach is employed to guide the FIB milling process of vitrified cellular samples and to capture specific structures, namely fluorescently labeled lipid droplets, in lamellas that are 300 nm thick. The correlation procedure is then applied to localize the fluorescently labeled structures in the transmission electron microscopy image of the lamella. This approach can be employed to navigate the acquisition of cryo-ET data within FIB-lamellas at specific locations, unambiguously identified by fluorescence microscopy.

  15. 3D printing method for freeform fabrication of optical phantoms simulating heterogeneous biological tissue

    NASA Astrophysics Data System (ADS)

    Wang, Minjie; Shen, Shuwei; Yang, Jie; Dong, Erbao; Xu, Ronald

    2014-03-01

    The performance of biomedical optical imaging devices heavily relies on appropriate calibration. However, many of existing calibration phantoms for biomedical optical devices are based on homogenous materials without considering the multi-layer heterogeneous structures observed in biological tissue. Using such a phantom for optical calibration may result in measurement bias. To overcome this problem, we propose a 3D printing method for freeform fabrication of tissue simulating phantoms with multilayer heterogeneous structure. The phantom simulates not only the morphologic characteristics of biological tissue but also absorption and scattering properties. The printing system is based on a 3D motion platform with coordinated control of the DC motors. A special jet nozzle is designed to mix base, scattering, and absorption materials at different ratios. 3D tissue structures are fabricated through layer-by-layer printing with selective deposition of phantom materials of different ingredients. Different mixed ratios of base, scattering and absorption materials have been tested in order to optimize the printing outcome. A spectrometer and a tissue spectrophotometer are used for characterizing phantom absorption and scattering properties. The goal of this project is to fabricate skin tissue simulating phantoms as a traceable standard for the calibration of biomedical optical spectral devices.

  16. PF2fit: Polar Fast Fourier Matched Alignment of Atomistic Structures with 3D Electron Microscopy Maps.

    PubMed

    Bettadapura, Radhakrishna; Rasheed, Muhibur; Vollrath, Antje; Bajaj, Chandrajit

    2015-10-01

    There continue to be increasing occurrences of both atomistic structure models in the PDB (possibly reconstructed from X-ray diffraction or NMR data), and 3D reconstructed cryo-electron microscopy (3D EM) maps (albeit at coarser resolution) of the same or homologous molecule or molecular assembly, deposited in the EMDB. To obtain the best possible structural model of the molecule at the best achievable resolution, and without any missing gaps, one typically aligns (match and fits) the atomistic structure model with the 3D EM map. We discuss a new algorithm and generalized framework, named PF(2) fit (Polar Fast Fourier Fitting) for the best possible structural alignment of atomistic structures with 3D EM. While PF(2) fit enables only a rigid, six dimensional (6D) alignment method, it augments prior work on 6D X-ray structure and 3D EM alignment in multiple ways: Scoring. PF(2) fit includes a new scoring scheme that, in addition to rewarding overlaps between the volumes occupied by the atomistic structure and 3D EM map, rewards overlaps between the volumes complementary to them. We quantitatively demonstrate how this new complementary scoring scheme improves upon existing approaches. PF(2) fit also includes two scoring functions, the non-uniform exterior penalty and the skeleton-secondary structure score, and implements the scattering potential score as an alternative to traditional Gaussian blurring. Search. PF(2) fit utilizes a fast polar Fourier search scheme, whose main advantage is the ability to search over uniformly and adaptively sampled subsets of the space of rigid-body motions. PF(2) fit also implements a new reranking search and scoring methodology that considerably improves alignment metrics in results obtained from the initial search.

  17. PF2 fit: Polar Fast Fourier Matched Alignment of Atomistic Structures with 3D Electron Microscopy Maps

    PubMed Central

    Bettadapura, Radhakrishna; Rasheed, Muhibur; Vollrath, Antje; Bajaj, Chandrajit

    2015-01-01

    There continue to be increasing occurrences of both atomistic structure models in the PDB (possibly reconstructed from X-ray diffraction or NMR data), and 3D reconstructed cryo-electron microscopy (3D EM) maps (albeit at coarser resolution) of the same or homologous molecule or molecular assembly, deposited in the EMDB. To obtain the best possible structural model of the molecule at the best achievable resolution, and without any missing gaps, one typically aligns (match and fits) the atomistic structure model with the 3D EM map. We discuss a new algorithm and generalized framework, named PF2 fit (Polar Fast Fourier Fitting) for the best possible structural alignment of atomistic structures with 3D EM. While PF2 fit enables only a rigid, six dimensional (6D) alignment method, it augments prior work on 6D X-ray structure and 3D EM alignment in multiple ways: Scoring. PF2 fit includes a new scoring scheme that, in addition to rewarding overlaps between the volumes occupied by the atomistic structure and 3D EM map, rewards overlaps between the volumes complementary to them. We quantitatively demonstrate how this new complementary scoring scheme improves upon existing approaches. PF2 fit also includes two scoring functions, the non-uniform exterior penalty and the skeleton-secondary structure score, and implements the scattering potential score as an alternative to traditional Gaussian blurring. Search. PF2 fit utilizes a fast polar Fourier search scheme, whose main advantage is the ability to search over uniformly and adaptively sampled subsets of the space of rigid-body motions. PF2 fit also implements a new reranking search and scoring methodology that considerably improves alignment metrics in results obtained from the initial search. PMID:26469938

  18. PF2fit: Polar Fast Fourier Matched Alignment of Atomistic Structures with 3D Electron Microscopy Maps.

    PubMed

    Bettadapura, Radhakrishna; Rasheed, Muhibur; Vollrath, Antje; Bajaj, Chandrajit

    2015-10-01

    There continue to be increasing occurrences of both atomistic structure models in the PDB (possibly reconstructed from X-ray diffraction or NMR data), and 3D reconstructed cryo-electron microscopy (3D EM) maps (albeit at coarser resolution) of the same or homologous molecule or molecular assembly, deposited in the EMDB. To obtain the best possible structural model of the molecule at the best achievable resolution, and without any missing gaps, one typically aligns (match and fits) the atomistic structure model with the 3D EM map. We discuss a new algorithm and generalized framework, named PF(2) fit (Polar Fast Fourier Fitting) for the best possible structural alignment of atomistic structures with 3D EM. While PF(2) fit enables only a rigid, six dimensional (6D) alignment method, it augments prior work on 6D X-ray structure and 3D EM alignment in multiple ways: Scoring. PF(2) fit includes a new scoring scheme that, in addition to rewarding overlaps between the volumes occupied by the atomistic structure and 3D EM map, rewards overlaps between the volumes complementary to them. We quantitatively demonstrate how this new complementary scoring scheme improves upon existing approaches. PF(2) fit also includes two scoring functions, the non-uniform exterior penalty and the skeleton-secondary structure score, and implements the scattering potential score as an alternative to traditional Gaussian blurring. Search. PF(2) fit utilizes a fast polar Fourier search scheme, whose main advantage is the ability to search over uniformly and adaptively sampled subsets of the space of rigid-body motions. PF(2) fit also implements a new reranking search and scoring methodology that considerably improves alignment metrics in results obtained from the initial search. PMID:26469938

  19. Sample holder for axial rotation of specimens in 3D microscopy.

    PubMed

    Bruns, T; Schickinger, S; Schneckenburger, H

    2015-10-01

    In common light microscopy, observation of samples is only possible from one perspective. However, especially for larger three-dimensional specimens observation from different views is desirable. Therefore, we are presenting a sample holder permitting rotation of the specimen around an axis perpendicular to the light path of the microscope. Thus, images can be put into a defined multidimensional context, enabling reliable three-dimensional reconstructions. The device can be easily adapted to a great variety of common light microscopes and is suitable for various applications in science, education and industry, where the observation of three-dimensional specimens is essential. Fluorescence z-projection images of copepods and ixodidae ticks at different rotation angles obtained by confocal laser scanning microscopy and light sheet fluorescence microscopy are reported as representative results.

  20. Multi-modal digital holographic microscopy for wide-field fluorescence and 3D phase imaging

    NASA Astrophysics Data System (ADS)

    Quan, Xiangyu; Xia, Peng; Matoba, Osamu; Nitta, Koichi; Awatsuji, Yasuhiro

    2016-03-01

    Multi-modal digital holographic microscopy is a combination of epifluorescence microscopy and digital holographic microscopy, the main function of which is to obtain images from fluorescence intensity and quantified phase contrasts, simultaneously. The proposed system is mostly beneficial to biological studies, with the reason that often the studies are depending on fluorescent labeling techniques to detect certain intracellular molecules, while phase information reflecting properties of unstained transparent elements. This paper is presenting our latest researches on applications such as randomly moving micro-fluorescent beads and living cells of Physcomitrella patens. The experiments are succeeded on obtaining a succession of wide-field fluorescent images and holograms from micro-beads, and different depths focusing is realized via numerical reconstruction. Living cells of Physcomitrella patens are recorded in the static manner, the reconstruction distance indicates thickness of cellular structure. These results are implementing practical applications toward many biomedical science researches.

  1. High-resolution high-sensitivity elemental imaging by secondary ion mass spectrometry: from traditional 2D and 3D imaging to correlative microscopy

    NASA Astrophysics Data System (ADS)

    Wirtz, T.; Philipp, P.; Audinot, J.-N.; Dowsett, D.; Eswara, S.

    2015-10-01

    Secondary ion mass spectrometry (SIMS) constitutes an extremely sensitive technique for imaging surfaces in 2D and 3D. Apart from its excellent sensitivity and high lateral resolution (50 nm on state-of-the-art SIMS instruments), advantages of SIMS include high dynamic range and the ability to differentiate between isotopes. This paper first reviews the underlying principles of SIMS as well as the performance and applications of 2D and 3D SIMS elemental imaging. The prospects for further improving the capabilities of SIMS imaging are discussed. The lateral resolution in SIMS imaging when using the microprobe mode is limited by (i) the ion probe size, which is dependent on the brightness of the primary ion source, the quality of the optics of the primary ion column and the electric fields in the near sample region used to extract secondary ions; (ii) the sensitivity of the analysis as a reasonable secondary ion signal, which must be detected from very tiny voxel sizes and thus from a very limited number of sputtered atoms; and (iii) the physical dimensions of the collision cascade determining the origin of the sputtered ions with respect to the impact site of the incident primary ion probe. One interesting prospect is the use of SIMS-based correlative microscopy. In this approach SIMS is combined with various high-resolution microscopy techniques, so that elemental/chemical information at the highest sensitivity can be obtained with SIMS, while excellent spatial resolution is provided by overlaying the SIMS images with high-resolution images obtained by these microscopy techniques. Examples of this approach are given by presenting in situ combinations of SIMS with transmission electron microscopy (TEM), helium ion microscopy (HIM) and scanning probe microscopy (SPM).

  2. Non-destructive mapping of grain orientations in 3D by laboratory X-ray microscopy

    NASA Astrophysics Data System (ADS)

    McDonald, S. A.; Reischig, P.; Holzner, C.; Lauridsen, E. M.; Withers, P. J.; Merkle, A. P.; Feser, M.

    2015-10-01

    The ability to characterise crystallographic microstructure, non-destructively and in three-dimensions, is a powerful tool for understanding many aspects related to damage and deformation mechanisms in polycrystalline materials. To this end, the technique of X-ray diffraction contrast tomography (DCT) using monochromatic synchrotron and polychromatic laboratory X-ray sources has been shown to be capable of mapping crystal grains and their orientations non-destructively in 3D. Here we describe a novel laboratory-based X-ray DCT modality (LabDCT), enabling the wider accessibility of the DCT technique for routine use and in-depth studies of, for example, temporal changes in crystallographic grain structure non-destructively over time through ‘4D’ in situ time-lapse studies. The capability of the technique is demonstrated by studying a titanium alloy (Ti-β21S) sample. In the current implementation the smallest grains that can be reliably detected are around 40 μm. The individual grain locations and orientations are reconstructed using the LabDCT method and the results are validated against independent measurements from phase contrast tomography and electron backscatter diffraction respectively. Application of the technique promises to provide important insights related to the roles of recrystallization and grain growth on materials properties as well as supporting 3D polycrystalline modelling of materials performance.

  3. Non-destructive mapping of grain orientations in 3D by laboratory X-ray microscopy

    PubMed Central

    McDonald, S. A.; Reischig, P.; Holzner, C.; Lauridsen, E. M.; Withers, P. J.; Merkle, A. P.; Feser, M.

    2015-01-01

    The ability to characterise crystallographic microstructure, non-destructively and in three-dimensions, is a powerful tool for understanding many aspects related to damage and deformation mechanisms in polycrystalline materials. To this end, the technique of X-ray diffraction contrast tomography (DCT) using monochromatic synchrotron and polychromatic laboratory X-ray sources has been shown to be capable of mapping crystal grains and their orientations non-destructively in 3D. Here we describe a novel laboratory-based X-ray DCT modality (LabDCT), enabling the wider accessibility of the DCT technique for routine use and in-depth studies of, for example, temporal changes in crystallographic grain structure non-destructively over time through ‘4D’ in situ time-lapse studies. The capability of the technique is demonstrated by studying a titanium alloy (Ti-β21S) sample. In the current implementation the smallest grains that can be reliably detected are around 40 μm. The individual grain locations and orientations are reconstructed using the LabDCT method and the results are validated against independent measurements from phase contrast tomography and electron backscatter diffraction respectively. Application of the technique promises to provide important insights related to the roles of recrystallization and grain growth on materials properties as well as supporting 3D polycrystalline modelling of materials performance. PMID:26494523

  4. 3D Plant cell architecture of Arabidopsis thaliana (Brassicaceae) using focused ion beam–scanning electron microscopy1

    PubMed Central

    Bhawana; Miller, Joyce L.; Cahoon, A. Bruce

    2014-01-01

    • Premise of the study: Focused ion beam–scanning electron microscopy (FIB-SEM) combines the ability to sequentially mill the sample surface and obtain SEM images that can be used to create 3D renderings with micron-level resolution. We have applied FIB-SEM to study Arabidopsis cell architecture. The goal was to determine the efficacy of this technique in plant tissue and cellular studies and to demonstrate its usefulness in studying cell and organelle architecture and distribution. • Methods: Seed aleurone, leaf mesophyll, stem cortex, root cortex, and petal lamina from Arabidopsis were fixed and embedded for electron microscopy using protocols developed for animal tissues and modified for use with plant cells. Each sample was sectioned using the FIB and imaged with SEM. These serial images were assembled to produce 3D renderings of each cell type. • Results: Organelles such as nuclei and chloroplasts were easily identifiable, and other structures such as endoplasmic reticula, lipid bodies, and starch grains were distinguishable in each tissue. • Discussion: The application of FIB-SEM produced 3D renderings of five plant cell types and offered unique views of their shapes and internal content. These results demonstrate the usefulness of FIB-SEM for organelle distribution and cell architecture studies. PMID:25202629

  5. Confocal microscopy of thick tissue sections: 3D visualizaiton of rat kidney glomeruli

    EPA Science Inventory

    Confocal laser scanning microscopy (CLSM) as a technique capable of generating serial sections of whole-mount tissue and then reassembling the computer-acquired images as a virtual 3-dimentional structure. In many ways CLSM offers an alternative to traditional sectioning approac...

  6. Confocal Microscopy of thick tissue sections: 3D Visualization of rat kidney glomeruli

    EPA Science Inventory

    Confocal laser scanning microscopy (CLSM) as a technique capable of generating serial sections of whole-mount tissue and then reassembling the computer-acquired images as a virtual 3-dimentional structure. In many ways CLSM offers an alternative to traditional sectioning approac...

  7. Analysis of thin baked-on silicone layers by FTIR and 3D-Laser Scanning Microscopy.

    PubMed

    Funke, Stefanie; Matilainen, Julia; Nalenz, Heiko; Bechtold-Peters, Karoline; Mahler, Hanns-Christian; Friess, Wolfgang

    2015-10-01

    Pre-filled syringes (PFS) and auto-injection devices with cartridges are increasingly used for parenteral administration. To assure functionality, silicone oil is applied to the inner surface of the glass barrel. Silicone oil migration into the product can be minimized by applying a thin but sufficient layer of silicone oil emulsion followed by thermal bake-on versus spraying-on silicone oil. Silicone layers thicker than 100nm resulting from regular spray-on siliconization can be characterized using interferometric profilometers. However, the analysis of thin silicone layers generated by bake-on siliconization is more challenging. In this paper, we have evaluated Fourier transform infrared (FTIR) spectroscopy after solvent extraction and a new 3D-Laser Scanning Microscopy (3D-LSM) to overcome this challenge. A multi-step solvent extraction and subsequent FTIR spectroscopy enabled to quantify baked-on silicone levels as low as 21-325μg per 5mL cartridge. 3D-LSM was successfully established to visualize and measure baked-on silicone layers as thin as 10nm. 3D-LSM was additionally used to analyze the silicone oil distribution within cartridges at such low levels. Both methods provided new, highly valuable insights to characterize the siliconization after processing, in order to achieve functionality.

  8. 3D imaging and quantitative analysis of small solubilized membrane proteins and their complexes by transmission electron microscopy.

    PubMed

    Vahedi-Faridi, Ardeschir; Jastrzebska, Beata; Palczewski, Krzysztof; Engel, Andreas

    2013-02-01

    Inherently unstable, detergent-solubilized membrane protein complexes can often not be crystallized. For complexes that have a mass of >300 kDa, cryo-electron microscopy (EM) allows their three-dimensional (3D) structure to be assessed to a resolution that makes secondary structure elements visible in the best case. However, many interesting complexes exist whose mass is below 300 kDa and thus need alternative approaches. Two methods are reviewed: (i) Mass measurement in a scanning transmission electron microscope, which has provided important information on the stoichiometry of membrane protein complexes. This technique is applicable to particulate, filamentous and sheet-like structures. (ii) 3D-EM of negatively stained samples, which determines the molecular envelope of small membrane protein complexes. Staining and dehydration artifacts may corrupt the quality of the 3D map. Staining conditions thus need to be optimized. 3D maps of plant aquaporin SoPIP2;1 tetramers solubilized in different detergents illustrate that the flattening artifact can be partially prevented and that the detergent itself contributes significantly. Another example discussed is the complex of G protein-coupled receptor rhodopsin with its cognate G protein transducin.

  9. Parametric estimation of 3D tubular structures for diffuse optical tomography

    PubMed Central

    Larusson, Fridrik; Anderson, Pamela G.; Rosenberg, Elizabeth; Kilmer, Misha E.; Sassaroli, Angelo; Fantini, Sergio; Miller, Eric L.

    2013-01-01

    We explore the use of diffuse optical tomography (DOT) for the recovery of 3D tubular shapes representing vascular structures in breast tissue. Using a parametric level set method (PaLS) our method incorporates the connectedness of vascular structures in breast tissue to reconstruct shape and absorption values from severely limited data sets. The approach is based on a decomposition of the unknown structure into a series of two dimensional slices. Using a simplified physical model that ignores 3D effects of the complete structure, we develop a novel inter-slice regularization strategy to obtain global regularity. We report on simulated and experimental reconstructions using realistic optical contrasts where our method provides a more accurate estimate compared to an unregularized approach and a pixel based reconstruction. PMID:23411913

  10. Design and verification of diffractive optical elements for speckle generation of 3-D range sensors

    NASA Astrophysics Data System (ADS)

    Du, Pei-Qin; Shih, Hsi-Fu; Chen, Jenq-Shyong; Wang, Yi-Shiang

    2016-09-01

    The optical projection using speckles is one of the structured light methods that have been applied to three-dimensional (3-D) range sensors. This paper investigates the design and fabrication of diffractive optical elements (DOEs) for generating the light field with uniformly distributed speckles. Based on the principles of computer generated holograms, the iterative Fourier transform algorithm was adopted for the DOE design. It was used to calculate the phase map for diffracting the incident laser beam into a goal pattern with distributed speckles. Four patterns were designed in the study. Their phase maps were first examined by a spatial light modulator and then fabricated on glass substrates by microfabrication processes. Finally, the diffraction characteristics of the fabricated devices were verified. The experimental results show that the proposed methods are applicable to the DOE design of 3-D range sensors. Furthermore, any expected diffraction area and speckle density could be possibly achieved according to the relations presented in the paper.

  11. Visualising the 3D Structure of Fine-Grained Estuarine Sediments; Preliminary Interpretations of a Novel Dataset Obtained via Volume Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Wheatland, Jonathan; Bushby, Andy; Spencer, Kate; Carr, Simon

    2014-05-01

    Accurate measurement of the physical characteristics of sediment are critical to determining sediment transport behaviour and the stability of settled deposits. The properties (e.g. particle size, density, and settling velocity) of coarse-grained sediments (> 63 μm φ) can be easily characterised, hence their behaviour is relatively simple to predict and model. However, due to their small size and tendency to interact with their surrounding medium, the characteristics of fine sediments (< 63 μm φ) and their behaviour during transportation, deposition and consolidation is poorly understood. Recent studies have used correlative microscopy, a multi-method technique combining scanning confocal laser microscopy (SCLM), conventional optical microscopy (COM), and transmission electron microscopy (TEM), to characterise fine sediments at both the gross (> 1 μm) and sub-micron scale (Droppo et al., 1996). Whilst this technique has proven insightful, the measurement of geometric properties (e.g. the shape of primary particles and their spatial arrangement) can only be achieved by three-dimensional (3D) analysis and the scale of observation for e.g. TEM does not overlap with those techniques used to characterise sediments at larger scales (100s to 1000s microns) (e.g. video analysis). Volume electron microscopy [or focused ion beam scanning electron microscopy (FIB-SEM)] provides 3D analysis at scales of 10s to 1000s microns and though widely used in cell biology, has not been used to observe sediment. FIB-SEM requires samples that are vacuum stable and a key challenge will be to capture fragile, hydrated sediment samples whilst preserving their structural integrity. The aims of this work are therefore: 1) to modify preparation techniques currently used in cell biology for the stabilization of sedimentary materials; 2) to acquire 3D datasets for both fragile suspended sediments (flocs) and consolidated bed sediments and 3) to interpret the 3D structure of these samples. In

  12. Web-based volume slicer for 3D electron-microscopy data from EMDB.

    PubMed

    Salavert-Torres, José; Iudin, Andrii; Lagerstedt, Ingvar; Sanz-García, Eduardo; Kleywegt, Gerard J; Patwardhan, Ardan

    2016-05-01

    We describe the functionality and design of the Volume slicer - a web-based slice viewer for EMDB entries. This tool uniquely provides the facility to view slices from 3D EM reconstructions along the three orthogonal axes and to rapidly switch between them and navigate through the volume. We have employed multiple rounds of user-experience testing with members of the EM community to ensure that the interface is easy and intuitive to use and the information provided is relevant. The impetus to develop the Volume slicer has been calls from the EM community to provide web-based interactive visualisation of 2D slice data. This would be useful for quick initial checks of the quality of a reconstruction. Again in response to calls from the community, we plan to further develop the Volume slicer into a fully-fledged Volume browser that provides integrated visualisation of EMDB and PDB entries from the molecular to the cellular scale.

  13. Web-based volume slicer for 3D electron-microscopy data from EMDB.

    PubMed

    Salavert-Torres, José; Iudin, Andrii; Lagerstedt, Ingvar; Sanz-García, Eduardo; Kleywegt, Gerard J; Patwardhan, Ardan

    2016-05-01

    We describe the functionality and design of the Volume slicer - a web-based slice viewer for EMDB entries. This tool uniquely provides the facility to view slices from 3D EM reconstructions along the three orthogonal axes and to rapidly switch between them and navigate through the volume. We have employed multiple rounds of user-experience testing with members of the EM community to ensure that the interface is easy and intuitive to use and the information provided is relevant. The impetus to develop the Volume slicer has been calls from the EM community to provide web-based interactive visualisation of 2D slice data. This would be useful for quick initial checks of the quality of a reconstruction. Again in response to calls from the community, we plan to further develop the Volume slicer into a fully-fledged Volume browser that provides integrated visualisation of EMDB and PDB entries from the molecular to the cellular scale. PMID:26876163

  14. Fast 3D visualization of endogenous brain signals with high-sensitivity laser scanning photothermal microscopy

    PubMed Central

    Miyazaki, Jun; Iida, Tadatsune; Tanaka, Shinji; Hayashi-Takagi, Akiko; Kasai, Haruo; Okabe, Shigeo; Kobayashi, Takayoshi

    2016-01-01

    A fast, high-sensitivity photothermal microscope was developed by implementing a spatially segmented balanced detection scheme into a laser scanning microscope. We confirmed a 4.9 times improvement in signal-to-noise ratio in the spatially segmented balanced detection compared with that of conventional detection. The system demonstrated simultaneous bi-modal photothermal and confocal fluorescence imaging of transgenic mouse brain tissue with a pixel dwell time of 20 μs. The fluorescence image visualized neurons expressing yellow fluorescence proteins, while the photothermal signal detected endogenous chromophores in the mouse brain, allowing 3D visualization of the distribution of various features such as blood cells and fine structures probably due to lipids. This imaging modality was constructed using compact and cost-effective laser diodes, and will thus be widely useful in the life and medical sciences. PMID:27231615

  15. Uncertainty studies of topographical measurements on steel surface corrosion by 3D scanning electron microscopy.

    PubMed

    Kang, K W; Pereda, M D; Canafoglia, M E; Bilmes, P; Llorente, C; Bonetto, R

    2012-02-01

    Pitting corrosion is a damage mechanism quite serious and dangerous in both carbon steel boiler tubes for power plants which are vital to most industries and stainless steels for orthopedic human implants whose demand, due to the increase of life expectation and rate of traffic accidents, has sharply increased. Reliable methods to characterize this kind of damage are becoming increasingly necessary, when trying to evaluate the advance of damage and to establish the best procedures for component inspection in order to determine remaining lives and failure mitigation. A study about the uncertainties on the topographies of corrosion pits from 3D SEM images, obtained at low magnifications (where errors are greater) and different stage tilt angles were carried out using an in-house software previously developed. Additionally, measurements of pit depths on biomaterial surfaces, subjected to two different surface treatments on stainless steels, were carried out. The different depth distributions observed were in agreement with electrochemical measurements.

  16. 3D printed sensing patches with embedded polymer optical fibre Bragg gratings

    NASA Astrophysics Data System (ADS)

    Zubel, Michal G.; Sugden, Kate; Saez-Rodriguez, D.; Nielsen, K.; Bang, O.

    2016-05-01

    The first demonstration of a polymer optical fibre Bragg grating (POFBG) embedded in a 3-D printed structure is reported. Its cyclic strain performance and temperature characteristics are examined and discussed. The sensing patch has a repeatable strain sensitivity of 0.38 pm/μepsilon. Its temperature behaviour is unstable, with temperature sensitivity values varying between 30-40 pm/°C.

  17. Imaging and 3-D dosimetry: top tips for MRI and optical CT

    NASA Astrophysics Data System (ADS)

    Doran, Simon J.

    2010-11-01

    The conference "refresher session" associated with this abstract reviews the main principles of the two most important imaging readout modalities for 3-D dosimetry: MRI and optical CT. Best practices for both these techniques are already described in several different places in the literature, but, for the uninitiated, there are a number of pitfalls. Here, I list some of the important considerations required to obtain good results from these methods and point to relevant prior work.

  18. Rapid fabrication of complex 3D extracellular microenvironments by dynamic optical projection stereolithography.

    PubMed

    Zhang, A Ping; Qu, Xin; Soman, Pranav; Hribar, Kolin C; Lee, Jin W; Chen, Shaochen; He, Sailing

    2012-08-16

    The topographic features of the extracelluar matrix (ECM) lay the foundation for cellular behavior. A novel biofabrication method using a digital-mirror device (DMD), called dynamic optical projection stereolithography (DOPsL) is demonstrated. This robust and versatile platform can generate complex biomimetic scaffolds within seconds. Such 3D scaffolds have promising potentials for studying cell interactions with microenvironments in vitro and in vivo.

  19. Fast 3D optical reconstruction in turbid media using spatially modulated light

    PubMed Central

    D’Andrea, Cosimo; Ducros, Nicolas; Bassi, Andrea; Arridge, Simon; Valentini, Gianluca

    2010-01-01

    A method to perform fast 3-D optical reconstruction, based on structured light, in thick samples is demonstrated and experimentally validated. The experimental and reconstruction procedure, based on Finite Elements Method, used to reconstruct absorbing heterogeneities, with arbitrary arrangement in space, is discussed. In particular we demonstrated that a 2D sampling of the source Fourier plane is required to improve the imaging capability. PMID:21258482

  20. Viral Infection at High Magnification: 3D Electron Microscopy Methods to Analyze the Architecture of Infected Cells.

    PubMed

    Romero-Brey, Inés; Bartenschlager, Ralf

    2015-12-03

    As obligate intracellular parasites, viruses need to hijack their cellular hosts and reprogram their machineries in order to replicate their genomes and produce new virions. For the direct visualization of the different steps of a viral life cycle (attachment, entry, replication, assembly and egress) electron microscopy (EM) methods are extremely helpful. While conventional EM has given important information about virus-host cell interactions, the development of three-dimensional EM (3D-EM) approaches provides unprecedented insights into how viruses remodel the intracellular architecture of the host cell. During the last years several 3D-EM methods have been developed. Here we will provide a description of the main approaches and examples of innovative applications.

  1. Viral Infection at High Magnification: 3D Electron Microscopy Methods to Analyze the Architecture of Infected Cells

    PubMed Central

    Romero-Brey, Inés; Bartenschlager, Ralf

    2015-01-01

    As obligate intracellular parasites, viruses need to hijack their cellular hosts and reprogram their machineries in order to replicate their genomes and produce new virions. For the direct visualization of the different steps of a viral life cycle (attachment, entry, replication, assembly and egress) electron microscopy (EM) methods are extremely helpful. While conventional EM has given important information about virus-host cell interactions, the development of three-dimensional EM (3D-EM) approaches provides unprecedented insights into how viruses remodel the intracellular architecture of the host cell. During the last years several 3D-EM methods have been developed. Here we will provide a description of the main approaches and examples of innovative applications. PMID:26633469

  2. Performance of a commercial optical CT scanner and polymer gel dosimeters for 3-D dose verification

    SciTech Connect

    Xu, Y.; Wuu, C.-S.; Maryanski, Marek J.

    2004-11-01

    Performance analysis of a commercial three-dimensional (3-D) dose mapping system based on optical CT scanning of polymer gels is presented. The system consists of BANG{sup reg}3 polymer gels (MGS Research, Inc., Madison, CT), OCTOPUS{sup TM} laser CT scanner (MGS Research, Inc., Madison, CT), and an in-house developed software for optical CT image reconstruction and 3-D dose distribution comparison between the gel, film measurements and the radiation therapy treatment plans. Various sources of image noise (digitization, electronic, optical, and mechanical) generated by the scanner as well as optical uniformity of the polymer gel are analyzed. The performance of the scanner is further evaluated in terms of the reproducibility of the data acquisition process, the uncertainties at different levels of reconstructed optical density per unit length and the effects of scanning parameters. It is demonstrated that for BANG{sup registered}3 gel phantoms held in cylindrical plastic containers, the relative dose distribution can be reproduced by the scanner with an overall uncertainty of about 3% within approximately 75% of the radius of the container. In regions located closer to the container wall, however, the scanner generates erroneous optical density values that arise from the reflection and refraction of the laser rays at the interface between the gel and the container. The analysis of the accuracy of the polymer gel dosimeter is exemplified by the comparison of the gel/OCT-derived dose distributions with those from film measurements and a commercial treatment planning system (Cadplan, Varian Corporation, Palo Alto, CA) for a 6 cmx6 cm single field of 6 MV x rays and a 3-D conformal radiotherapy (3DCRT) plan. The gel measurements agree with the treatment plans and the film measurements within the '3%-or-2 mm' criterion throughout the usable, artifact-free central region of the gel volume. Discrepancies among the three data sets are analyzed.

  3. Isolation, Electron Microscopy and 3D Reconstruction of Invertebrate Muscle Myofilaments

    PubMed Central

    Craig, Roger

    2011-01-01

    Understanding the molecular mechanism of muscle contraction and its regulation has been greatly influenced and aided by studies of myofilament structure in invertebrate muscles. Invertebrates are easily obtained and cover a broad spectrum of species and functional specializations. The thick (myosin-containing) filaments from some invertebrates are especially stable and simple in structure and thus much more amenable to structural analysis than those of vertebrates. Comparative studies of invertebrate filaments by electron microscopy and image processing have provided important generalizations of muscle molecular structure and function. This article reviews methods for preparing thick and thin filaments from invertebrate muscle, for imaging filaments by electron microscopy, and for determining their three dimensional structure by image processing. It also highlights some of the key insights into filament function that have come from these studies. PMID:22155190

  4. Scanning transmission and computer-aided volumic electron microscopy: 3-D modeling of entire cells by electronic imaging

    NASA Astrophysics Data System (ADS)

    Bron, Christophe; Gremillet, Philip; Launay, D.; Jourlin, Michel; Gautschi, H. P.; Baechi, Thomas; Schuepbach, Joerg

    1990-05-01

    The digital processing of electron microscopic images from serial sections containing laser-induced topographical references allows a 3-D reconstruction at a depth resolution of 30 to 40 nm of entire cells by the use of image analysis methods, as already demonstrated for Transmission Electron Microscopy (TEM) coupled with a video camera. We decided to use a Scanning Transmission Electron Microscope (STEM) to get higher contrast and better resolution at medium magnification. The scanning of our specimens at video frequencies is an attractive and easy way to link a STEM with an image processing system but the hysteresis of the electronic spools responsible for the magnetic deviation of the scanning electron beam induces deformations of images which have to be modelized and corrected before registration. Computer algorithms developed for image analysis and treatment correct the artifacts caused by the use of STEM and by serial sectioning to automatically reconstruct the third dimension of the cells. They permit the normalization of the images through logarithmic processing of the original grey level infonnation. The automatic extraction of cell limits allows to link the image analysis and treatments with image synthesis methods by minimal human intervention. The surface representation and the registered images provide an ultrastructural data base from which quantitative 3-D morphological parameters, as well as otherwise impossible visualizations, can be computed. This 3-D image processing named C.A.V.U.M. for Computer Aided Volumic Ultra-Microscopy offers a new tool for the documentation and analysis of cell ultrastructure and for 3-D morphometric studies at EM magnifications. Further, a virtual observer can be computed in such a way as to simulate a visit of the reconstructed object.

  5. From Voxels to Knowledge: A Practical Guide to the Segmentation of Complex Electron Microscopy 3D-Data

    PubMed Central

    Tsai, Wen-Ting; Hassan, Ahmed; Sarkar, Purbasha; Correa, Joaquin; Metlagel, Zoltan; Jorgens, Danielle M.; Auer, Manfred

    2014-01-01

    Modern 3D electron microscopy approaches have recently allowed unprecedented insight into the 3D ultrastructural organization of cells and tissues, enabling the visualization of large macromolecular machines, such as adhesion complexes, as well as higher-order structures, such as the cytoskeleton and cellular organelles in their respective cell and tissue context. Given the inherent complexity of cellular volumes, it is essential to first extract the features of interest in order to allow visualization, quantification, and therefore comprehension of their 3D organization. Each data set is defined by distinct characteristics, e.g., signal-to-noise ratio, crispness (sharpness) of the data, heterogeneity of its features, crowdedness of features, presence or absence of characteristic shapes that allow for easy identification, and the percentage of the entire volume that a specific region of interest occupies. All these characteristics need to be considered when deciding on which approach to take for segmentation. The six different 3D ultrastructural data sets presented were obtained by three different imaging approaches: resin embedded stained electron tomography, focused ion beam- and serial block face- scanning electron microscopy (FIB-SEM, SBF-SEM) of mildly stained and heavily stained samples, respectively. For these data sets, four different segmentation approaches have been applied: (1) fully manual model building followed solely by visualization of the model, (2) manual tracing segmentation of the data followed by surface rendering, (3) semi-automated approaches followed by surface rendering, or (4) automated custom-designed segmentation algorithms followed by surface rendering and quantitative analysis. Depending on the combination of data set characteristics, it was found that typically one of these four categorical approaches outperforms the others, but depending on the exact sequence of criteria, more than one approach may be successful. Based on these data

  6. Simultaneous electrochemical and 3D optical imaging of silver nanoparticle oxidation

    NASA Astrophysics Data System (ADS)

    Batchelor-McAuley, Christopher; Martinez-Marrades, Ariadna; Tschulik, Kristina; Patel, Anisha N.; Combellas, Catherine; Kanoufi, Frédéric; Tessier, Gilles; Compton, Richard G.

    2014-03-01

    The oxidation of AgNPs at a thin-film gold electrode is simultaneously investigated via digital holography and electrochemistry. The use of holography allows, for the first time, the 3D visualization of the electrochemical interfacial region at a relatively high acquisition rate. It is demonstrated how the coupling of these two techniques provides complementary chemical information. The ensemble response of the oxidation of surface-adsorbed silver nanoparticles to AgCl is monitored electrochemically, whereas this process is difficult to observe optically. Conversely, the subsequent chemical dissolution of individual AgCl nanocrystals can be tracked optically due to the associated decrease in the scattered light intensity.

  7. Characterization of a parallel beam CCD optical-CT apparatus for 3D radiation dosimetry

    NASA Astrophysics Data System (ADS)

    Krstajić, Nikola; Doran, Simon J.

    2006-12-01

    This paper describes the initial steps we have taken in establishing CCD based optical-CT as a viable alternative for 3-D radiation dosimetry. First, we compare the optical density (OD) measurements from a high quality test target and variable neutral density filter (VNDF). A modulation transfer function (MTF) of individual projections is derived for three positions of the sinusoidal test target within the scanning tank. Our CCD is then characterized in terms of its signal-to-noise ratio (SNR). Finally, a sample reconstruction of a scan of a PRESAGETM (registered trademark of Heuris Pharma, NJ, Skillman, USA.) dosimeter is given, demonstrating the capabilities of the apparatus.

  8. Optically clearing tissue as an initial step for 3D imaging of core biopsies to diagnose pancreatic cancer

    NASA Astrophysics Data System (ADS)

    Das, Ronnie; Agrawal, Aishwarya; Upton, Melissa P.; Seibel, Eric J.

    2014-02-01

    The pancreas is a deeply seated organ requiring endoscopically, or radiologically guided biopsies for tissue diagnosis. Current approaches include either fine needle aspiration biopsy (FNA) for cytologic evaluation, or core needle biopsies (CBs), which comprise of tissue cores (L = 1-2 cm, D = 0.4-2.0 mm) for examination by brightfield microscopy. Between procurement and visualization, biospecimens must be processed, sectioned and mounted on glass slides for 2D visualization. Optical information about the native tissue state can be lost with each procedural step and a pathologist cannot appreciate 3D organization from 2D observations of tissue sections 1-8 μm in thickness. Therefore, how might histological disease assessment improve if entire, intact CBs could be imaged in both brightfield and 3D? CBs are mechanically delicate; therefore, a simple device was made to cut intact, simulated CBs (L = 1-2 cm, D = 0.2-0.8 mm) from porcine pancreas. After CBs were laid flat in a chamber, z-stack images at 20x and 40x were acquired through the sample with and without the application of an optical clearing agent (FocusClear®). Intensity of transmitted light increased by 5-15x and islet structures unique to pancreas were clearly visualized 250-300 μm beneath the tissue surface. CBs were then placed in index matching square capillary tubes filled with FocusClear® and a standard optical clearing agent. Brightfield z-stack images were then acquired to present 3D visualization of the CB to the pathologist.

  9. Minimizing camera-eye optical aberrations during the 3D reconstruction of retinal structures

    NASA Astrophysics Data System (ADS)

    Aldana-Iuit, Javier; Martinez-Perez, M. Elena; Espinosa-Romero, Arturo; Diaz-Uribe, Rufino

    2010-05-01

    3D reconstruction of blood vessels is a powerful visualization tool for physicians, since it allows them to refer to qualitative representation of their subject of study. In this paper we propose a 3D reconstruction method of retinal vessels from fundus images. The reconstruction method propose herein uses images of the same retinal structure in epipolar geometry. Images are preprocessed by RISA system for segmenting blood vessels and obtaining feature points for correspondences. The correspondence points process is solved using correlation. The LMedS analysis and Graph Transformation Matching algorithm are used for outliers suppression. Camera projection matrices are computed with the normalized eight point algorithm. Finally, we retrieve 3D position of the retinal tree points by linear triangulation. In order to increase the power of visualization, 3D tree skeletons are represented by surfaces via generalized cylinders whose radius correspond to morphological measurements obtained by RISA. In this paper the complete calibration process including the fundus camera and the optical properties of the eye, the so called camera-eye system is proposed. On one hand, the internal parameters of the fundus camera are obtained by classical algorithms using a reference pattern. On the other hand, we minimize the undesirable efects of the aberrations induced by the eyeball optical system assuming that contact enlarging lens corrects astigmatism, spherical and coma aberrations are reduced changing the aperture size and eye refractive errors are suppressed adjusting camera focus during image acquisition. Evaluation of two self-calibration proposals and results of 3D blood vessel surface reconstruction are presented.

  10. Active optical system for advanced 3D surface structuring by laser remelting

    NASA Astrophysics Data System (ADS)

    Pütsch, O.; Temmler, A.; Stollenwerk, J.; Willenborg, E.; Loosen, P.

    2015-03-01

    Structuring by laser remelting enables completely new possibilities for designing surfaces since material is redistributed but not wasted. In addition to technological advantages, cost and time benefits yield from shortened process times, the avoidance of harmful chemicals and the elimination of subsequent finishing steps such as cleaning and polishing. The functional principle requires a completely new optical machine technology that maintains the spatial and temporal superposition and manipulation of three different laser beams emitted from two laser sources of different wavelength. The optical system has already been developed and demonstrated for the processing of flat samples of hot and cold working steel. However, since particularly the structuring of 3D-injection molds represents an application example of high innovation potential, the optical system has to take into account the elliptical beam geometry that occurs when the laser beams irradiate a curved surface. To take full advantage of structuring by remelting for the processing of 3D surfaces, additional optical functionality, called EPS (elliptical pre-shaping) has to be integrated into the existing set-up. The development of the beam shaping devices not only requires the analysis of the mechanisms of the beam projection but also a suitable optical design. Both aspects are discussed in this paper.

  11. TeraStitcher - A tool for fast automatic 3D-stitching of teravoxel-sized microscopy images

    PubMed Central

    2012-01-01

    Background Further advances in modern microscopy are leading to teravoxel-sized tiled 3D images at high resolution, thus increasing the dimension of the stitching problem of at least two orders of magnitude. The existing software solutions do not seem adequate to address the additional requirements arising from these datasets, such as the minimization of memory usage and the need to process just a small portion of data. Results We propose a free and fully automated 3D Stitching tool designed to match the special requirements coming out of teravoxel-sized tiled microscopy images that is able to stitch them in a reasonable time even on workstations with limited resources. The tool was tested on teravoxel-sized whole mouse brain images with micrometer resolution and it was also compared with the state-of-the-art stitching tools on megavoxel-sized publicy available datasets. This comparison confirmed that the solutions we adopted are suited for stitching very large images and also perform well on datasets with different characteristics. Indeed, some of the algorithms embedded in other stitching tools could be easily integrated in our framework if they turned out to be more effective on other classes of images. To this purpose, we designed a software architecture which separates the strategies that use efficiently memory resources from the algorithms which may depend on the characteristics of the acquired images. Conclusions TeraStitcher is a free tool that enables the stitching of Teravoxel-sized tiled microscopy images even on workstations with relatively limited resources of memory (<8 GB) and processing power. It exploits the knowledge of approximate tile positions and uses ad-hoc strategies and algorithms designed for such very large datasets. The produced images can be saved into a multiresolution representation to be efficiently retrieved and processed. We provide TeraStitcher both as standalone application and as plugin of the free software Vaa3D. PMID:23181553

  12. A joint estimation detection of Glaucoma progression in 3D spectral domain optical coherence tomography optic nerve head images

    NASA Astrophysics Data System (ADS)

    Belghith, Akram; Bowd, Christopher; Weinreb, Robert N.; Zangwill, Linda M.

    2014-03-01

    Glaucoma is an ocular disease characterized by distinctive changes in the optic nerve head (ONH) and visual field. Glaucoma can strike without symptoms and causes blindness if it remains without treatment. Therefore, early disease detection is important so that treatment can be initiated and blindness prevented. In this context, important advances in technology for non-invasive imaging of the eye have been made providing quantitative tools to measure structural changes in ONH topography, an essential element for glaucoma detection and monitoring. 3D spectral domain optical coherence tomography (SD-OCT), an optical imaging technique, has been commonly used to discriminate glaucomatous from healthy subjects. In this paper, we present a new framework for detection of glaucoma progression using 3D SD-OCT images. In contrast to previous works that the retinal nerve fiber layer (RNFL) thickness measurement provided by commercially available spectral-domain optical coherence tomograph, we consider the whole 3D volume for change detection. To integrate a priori knowledge and in particular the spatial voxel dependency in the change detection map, we propose the use of the Markov Random Field to handle a such dependency. To accommodate the presence of false positive detection, the estimated change detection map is then used to classify a 3D SDOCT image into the "non-progressing" and "progressing" glaucoma classes, based on a fuzzy logic classifier. We compared the diagnostic performance of the proposed framework to existing methods of progression detection.

  13. A Quantitative 3D Motility Analysis of Trypanosoma brucei by Use of Digital In-line Holographic Microscopy

    PubMed Central

    Weiße, Sebastian; Heddergott, Niko; Heydt, Matthias; Pflästerer, Daniel; Maier, Timo; Haraszti, Tamás; Grunze, Michael; Engstler, Markus; Rosenhahn, Axel

    2012-01-01

    We present a quantitative 3D analysis of the motility of the blood parasite Trypanosoma brucei. Digital in-line holographic microscopy has been used to track single cells with high temporal and spatial accuracy to obtain quantitative data on their behavior. Comparing bloodstream form and insect form trypanosomes as well as mutant and wildtype cells under varying external conditions we were able to derive a general two-state-run-and-tumble-model for trypanosome motility. Differences in the motility of distinct strains indicate that adaption of the trypanosomes to their natural environments involves a change in their mode of swimming. PMID:22629379

  14. EMRinger: side chain–directed model and map validation for 3D cryo-electron microscopy

    DOE PAGES

    Barad, Benjamin A.; Echols, Nathaniel; Wang, Ray Yu-Ruei; Cheng, Yifan; DiMaio, Frank; Adams, Paul D.; Fraser, James S.

    2015-08-17

    Advances in high-resolution cryo-electron microscopy (cryo-EM) require the development of validation metrics to independently assess map quality and model geometry. We report that EMRinger is a tool that assesses the precise fitting of an atomic model into the map during refinement and shows how radiation damage alters scattering from negatively charged amino acids. EMRinger (https://github.com/fraser-lab/EMRinger) will be useful for monitoring progress in resolving and modeling high-resolution features in cryo-EM.

  15. High-content 3D multicolor super-resolution localization microscopy.

    PubMed

    Pereira, Pedro M; Almada, Pedro; Henriques, Ricardo

    2015-01-01

    Super-resolution (SR) methodologies permit the visualization of cellular structures at near-molecular scale (1-30 nm), enabling novel mechanistic analysis of key events in cell biology not resolvable by conventional fluorescence imaging (∼300-nm resolution). When this level of detail is combined with computing power and fast and reliable analysis software, high-content screenings using SR becomes a practical option to address multiple biological questions. The importance of combining these powerful analytical techniques cannot be ignored, as they can address phenotypic changes on the molecular scale and in a statistically robust manner. In this work, we suggest an easy-to-implement protocol that can be applied to set up a high-content 3D SR experiment with user-friendly and freely available software. The protocol can be divided into two main parts: chamber and sample preparation, where a protocol to set up a direct STORM (dSTORM) sample is presented; and a second part where a protocol for image acquisition and analysis is described. We intend to take the reader step-by-step through the experimental process highlighting possible experimental bottlenecks and possible improvements based on recent developments in the field.

  16. Web-based volume slicer for 3D electron-microscopy data from EMDB

    PubMed Central

    Salavert-Torres, José; Iudin, Andrii; Lagerstedt, Ingvar; Sanz-García, Eduardo; Kleywegt, Gerard J.; Patwardhan, Ardan

    2016-01-01

    We describe the functionality and design of the Volume slicer – a web-based slice viewer for EMDB entries. This tool uniquely provides the facility to view slices from 3D EM reconstructions along the three orthogonal axes and to rapidly switch between them and navigate through the volume. We have employed multiple rounds of user-experience testing with members of the EM community to ensure that the interface is easy and intuitive to use and the information provided is relevant. The impetus to develop the Volume slicer has been calls from the EM community to provide web-based interactive visualisation of 2D slice data. This would be useful for quick initial checks of the quality of a reconstruction. Again in response to calls from the community, we plan to further develop the Volume slicer into a fully-fledged Volume browser that provides integrated visualisation of EMDB and PDB entries from the molecular to the cellular scale. PMID:26876163

  17. High-content 3D multicolor super-resolution localization microscopy.

    PubMed

    Pereira, Pedro M; Almada, Pedro; Henriques, Ricardo

    2015-01-01

    Super-resolution (SR) methodologies permit the visualization of cellular structures at near-molecular scale (1-30 nm), enabling novel mechanistic analysis of key events in cell biology not resolvable by conventional fluorescence imaging (∼300-nm resolution). When this level of detail is combined with computing power and fast and reliable analysis software, high-content screenings using SR becomes a practical option to address multiple biological questions. The importance of combining these powerful analytical techniques cannot be ignored, as they can address phenotypic changes on the molecular scale and in a statistically robust manner. In this work, we suggest an easy-to-implement protocol that can be applied to set up a high-content 3D SR experiment with user-friendly and freely available software. The protocol can be divided into two main parts: chamber and sample preparation, where a protocol to set up a direct STORM (dSTORM) sample is presented; and a second part where a protocol for image acquisition and analysis is described. We intend to take the reader step-by-step through the experimental process highlighting possible experimental bottlenecks and possible improvements based on recent developments in the field. PMID:25640426

  18. Near-wall 3D velocity measurements above biomimetic shark skin denticles using Digital In-line Holographic Microscopy

    NASA Astrophysics Data System (ADS)

    Toloui, Mostafa; Brajkovic, David; Hong, Jiarong

    2014-11-01

    Digital In-line Holography is employed to image 3D flow structures in the vicinity of a transparent rough surface consisting of closely packed biomimetic shark skin denticles as roughness elements. The 3D printed surface replicates the morphological features of real shark skin, and the denticles have a geometrical scale of 2 mm, i.e. 10 times of the real ones. In order to minimize optical aberrations near the fluid-roughness interface and enable flow measurements around denticles, the optical refractive index of the fluid medium is maintained the same as that of the denticle model in an index-matched flow facility using NaI solution as the working fluid. The experiment is conducted in a 1.2 m long test section with 50 mm × 50 mm cross section. The sampling volume is located in the downstream region of a shark skin replica of 12'' stretch where the turbulent flow is fully-developed and the transitional effect from smooth to the rough surface becomes negligible. Several instantaneous realizations of the 3D velocity field are obtained and are used to illustrate turbulent coherent structures induced by shark-skin denticles. This information will provide insights on the hydrodynamic function of shark's unique surface ornamentation.

  19. Stereoscopic 3D display with dynamic optical correction for recovering from asthenopia

    NASA Astrophysics Data System (ADS)

    Shibata, Takashi; Kawai, Takashi; Otsuki, Masaki; Miyake, Nobuyuki; Yoshihara, Yoshihiro; Iwasaki, Tsuneto

    2005-03-01

    The purpose of this study was to consider a practical application of a newly developed stereoscopic 3-D display that solves the problem of discrepancy between accommodation and convergence. The display uses dynamic optical correction to reduce the discrepancy, and can present images as if they are actually remote objects. The authors thought the display may assist in recovery from asthenopia, which is often caused when the eyes focus on a nearby object for a long time, such as in VDT (Visual Display Terminal) work. In general, recovery from asthenopia, and especially accommodative asthenopia, is achieved by focusing on distant objects. In order to verify this hypothesis, the authors performed visual acuity tests using Landolt rings before and after presenting stereoscopic 3-D images, and evaluated the degree of recovery from asthenopia. The experiment led to three main conclusions: (1) Visual acuity rose after viewing stereoscopic 3-D images on the developed display. (2) Recovery from asthenopia was particularly effective for the dominant eye in comparison with the other eye. (3) Interviews with the subjects indicated that the Landolt rings were particularly clear after viewing the stereoscopic 3-D images.

  20. Examination of asthenopia recovery using stereoscopic 3D display with dynamic optical correction

    NASA Astrophysics Data System (ADS)

    Shibata, Takashi; Kawai, Takashi; Ohta, Keiji; Lee, JaeLin; Otsuki, Masaki; Miyake, Nobuyuki; Yoshihara, Yoshihiro; Iwasaki, Tsuneto

    2006-02-01

    A common cause of asthenopia is viewing objects from a short distance, as is the case when working at a VDT (Visual Display Terminal). In general, recovery from asthenopia, especially accommodative asthenopia, is aided by looking into the distance. The authors have developed a stereoscopic 3-D display with dynamic optical correction that may reduce asthenopia. The display does this by reducing the discrepancy between accommodation and convergence, thereby presenting images as if they were actually in the distance. The results of visual acuity tests given before and after presenting stereoscopic 3-D images with this display show a tendency towards less asthenopia. In this study, the authors developed a refraction feedback function that makes the viewer's distance vision more effective when viewing stereoscopic 3-D images on the this display. Using this function, refraction is fed back during viewing and the viewer gradually acquires distance vision. The results of the study suggest that stereoscopic 3-D images are more effective than 2-D images for recovery from asthenopia.

  1. A comprehensive evaluation of the PRESAGE∕optical-CT 3D dosimetry system

    PubMed Central

    Sakhalkar, H. S.; Adamovics, J.; Ibbott, G.; Oldham, M.

    2009-01-01

    This work presents extensive investigations to evaluate the robustness (intradosimeter consistency and temporal stability of response), reproducibility, precision, and accuracy of a relatively new 3D dosimetry system comprising a leuco-dye doped plastic 3D dosimeter (PRESAGE) and a commercial optical-CT scanner (OCTOPUS 5× scanner from MGS Research, Inc). Four identical PRESAGE 3D dosimeters were created such that they were compatible with the Radiologic Physics Center (RPC) head-and-neck (H&N) IMRT credentialing phantom. Each dosimeter was irradiated with a rotationally symmetric arrangement of nine identical small fields (1×3 cm2) impinging on the flat circular face of the dosimeter. A repetitious sequence of three dose levels (4, 2.88, and 1.28 Gy) was delivered. The rotationally symmetric treatment resulted in a dose distribution with high spatial variation in axial planes but only gradual variation with depth along the long axis of the dosimeter. The significance of this treatment was that it facilitated accurate film dosimetry in the axial plane, for independent verification. Also, it enabled rigorous evaluation of robustness, reproducibility and accuracy of response, at the three dose levels. The OCTOPUS 5× commercial scanner was used for dose readout from the dosimeters at daily time intervals. The use of improved optics and acquisition technique yielded substantially improved noise characteristics (reduced to ∼2%) than has been achieved previously. Intradosimeter uniformity of radiochromic response was evaluated by calculating a 3D gamma comparison between each dosimeter and axially rotated copies of the same dosimeter. This convenient technique exploits the rotational symmetry of the distribution. All points in the gamma comparison passed a 2% difference, 1 mm distance-to-agreement criteria indicating excellent intradosimeter uniformity even at low dose levels. Postirradiation, the dosimeters were all found to exhibit a slight increase in opaqueness

  2. A comprehensive evaluation of the PRESAGE/optical-CT 3D dosimetry system.

    PubMed

    Sakhalkar, H S; Adamovics, J; Ibbott, G; Oldham, M

    2009-01-01

    This work presents extensive investigations to evaluate the robustness (intradosimeter consistency and temporal stability of response), reproducibility, precision, and accuracy of a relatively new 3D dosimetry system comprising a leuco-dye doped plastic 3D dosimeter (PRESAGE) and a commercial optical-CT scanner (OCTOPUS 5x scanner from MGS Research, Inc). Four identical PRESAGE 3D dosimeters were created such that they were compatible with the Radiologic Physics Center (RPC) head-and-neck (H&N) IMRT credentialing phantom. Each dosimeter was irradiated with a rotationally symmetric arrangement of nine identical small fields (1 x 3 cm2) impinging on the flat circular face of the dosimeter. A repetitious sequence of three dose levels (4, 2.88, and 1.28 Gy) was delivered. The rotationally symmetric treatment resulted in a dose distribution with high spatial variation in axial planes but only gradual variation with depth along the long axis of the dosimeter. The significance of this treatment was that it facilitated accurate film dosimetry in the axial plane, for independent verification. Also, it enabled rigorous evaluation of robustness, reproducibility and accuracy of response, at the three dose levels. The OCTOPUS 5x commercial scanner was used for dose readout from the dosimeters at daily time intervals. The use of improved optics and acquisition technique yielded substantially improved noise characteristics (reduced to approximately 2%) than has been achieved previously. Intradosimeter uniformity of radiochromic response was evaluated by calculating a 3D gamma comparison between each dosimeter and axially rotated copies of the same dosimeter. This convenient technique exploits the rotational symmetry of the distribution. All points in the gamma comparison passed a 2% difference, 1 mm distance-to-agreement criteria indicating excellent intradosimeter uniformity even at low dose levels. Postirradiation, the dosimeters were all found to exhibit a slight increase in

  3. A comprehensive evaluation of the PRESAGE/optical-CT 3D dosimetry system

    SciTech Connect

    Sakhalkar, H. S.; Adamovics, J.; Ibbott, G.; Oldham, M.

    2009-01-15

    This work presents extensive investigations to evaluate the robustness (intradosimeter consistency and temporal stability of response), reproducibility, precision, and accuracy of a relatively new 3D dosimetry system comprising a leuco-dye doped plastic 3D dosimeter (PRESAGE) and a commercial optical-CT scanner (OCTOPUS 5x scanner from MGS Research, Inc). Four identical PRESAGE 3D dosimeters were created such that they were compatible with the Radiologic Physics Center (RPC) head-and-neck (H and N) IMRT credentialing phantom. Each dosimeter was irradiated with a rotationally symmetric arrangement of nine identical small fields (1x3 cm{sup 2}) impinging on the flat circular face of the dosimeter. A repetitious sequence of three dose levels (4, 2.88, and 1.28 Gy) was delivered. The rotationally symmetric treatment resulted in a dose distribution with high spatial variation in axial planes but only gradual variation with depth along the long axis of the dosimeter. The significance of this treatment was that it facilitated accurate film dosimetry in the axial plane, for independent verification. Also, it enabled rigorous evaluation of robustness, reproducibility and accuracy of response, at the three dose levels. The OCTOPUS 5x commercial scanner was used for dose readout from the dosimeters at daily time intervals. The use of improved optics and acquisition technique yielded substantially improved noise characteristics (reduced to {approx}2%) than has been achieved previously. Intradosimeter uniformity of radiochromic response was evaluated by calculating a 3D gamma comparison between each dosimeter and axially rotated copies of the same dosimeter. This convenient technique exploits the rotational symmetry of the distribution. All points in the gamma comparison passed a 2% difference, 1 mm distance-to-agreement criteria indicating excellent intradosimeter uniformity even at low dose levels. Postirradiation, the dosimeters were all found to exhibit a slight increase in

  4. Master-slave interferometry for parallel spectral domain interferometry sensing and versatile 3D optical coherence tomography.

    PubMed

    Podoleanu, Adrian Gh; Bradu, Adrian

    2013-08-12

    Conventional spectral domain interferometry (SDI) methods suffer from the need of data linearization. When applied to optical coherence tomography (OCT), conventional SDI methods are limited in their 3D capability, as they cannot deliver direct en-face cuts. Here we introduce a novel SDI method, which eliminates these disadvantages. We denote this method as Master - Slave Interferometry (MSI), because a signal is acquired by a slave interferometer for an optical path difference (OPD) value determined by a master interferometer. The MSI method radically changes the main building block of an SDI sensor and of a spectral domain OCT set-up. The serially provided signal in conventional technology is replaced by multiple signals, a signal for each OPD point in the object investigated. This opens novel avenues in parallel sensing and in parallelization of signal processing in 3D-OCT, with applications in high- resolution medical imaging and microscopy investigation of biosamples. Eliminating the need of linearization leads to lower cost OCT systems and opens potential avenues in increasing the speed of production of en-face OCT images in comparison with conventional SDI.

  5. Efficient Semi-Automatic 3D Segmentation for Neuron Tracing in Electron Microscopy Images

    PubMed Central

    Jones, Cory; Liu, Ting; Cohan, Nathaniel Wood; Ellisman, Mark; Tasdizen, Tolga

    2015-01-01

    0.1. Background In the area of connectomics, there is a significant gap between the time required for data acquisition and dense reconstruction of the neural processes contained in the same dataset. Automatic methods are able to eliminate this timing gap, but the state-of-the-art accuracy so far is insufficient for use without user corrections. If completed naively, this process of correction can be tedious and time consuming. 0.2. New Method We present a new semi-automatic method that can be used to perform 3D segmentation of neurites in EM image stacks. It utilizes an automatic method that creates a hierarchical structure for recommended merges of superpixels. The user is then guided through each predicted region to quickly identify errors and establish correct links. 0.3. Results We tested our method on three datasets with both novice and expert users. Accuracy and timing were compared with published automatic, semi-automatic, and manual results. 0.4. Comparison with Existing Methods Post-automatic correction methods have also been used in [1] and [2]. These methods do not provide navigation or suggestions in the manner we present. Other semi-automatic methods require user input prior to the automatic segmentation such as [3] and [4] and are inherently different than our method. 0.5. Conclusion Using this method on the three datasets, novice users achieved accuracy exceeding state-of-the-art automatic results, and expert users achieved accuracy on par with full manual labeling but with a 70% time improvement when compared with other examples in publication. PMID:25769273

  6. STXM goes 3D: digital reconstruction of focal stacks as novel approach towards confocal soft x-ray microscopy.

    PubMed

    Späth, Andreas; Scho Ll, Simon; Riess, Christian; Schmidtel, Daniel; Paradossi, Gaio; Raabe, Jo Rg; Hornegger, Joachim; Fink, Rainer H

    2014-09-01

    Fresnel zone plate based soft x-ray transmission microspectroscopy has developed into a routine technique for high-resolution elemental or chemical 2D imaging of thin film specimens. The availability of high resolution Fresnel lenses with short depth of focus offers the possibility of optical slicing (in the third dimension) by focus series with resolutions in the submicron regime. We introduce a 3D reconstruction algorithm that uses a variance-based metric to assign a focus measure as basis for volume rendering. The algorithm is applied to simulated geometries and opaque soft matter specimens thus enabling 3D visualization. These studies with z-resolution of few 100nm serve as important step towards the vision of a confocal transmission x-ray microscope.

  7. 3D optical printing of piezoelectric nanoparticle-polymer composite materials.

    PubMed

    Kim, Kanguk; Zhu, Wei; Qu, Xin; Aaronson, Chase; McCall, William R; Chen, Shaochen; Sirbuly, Donald J

    2014-10-28

    Here we demonstrate that efficient piezoelectric nanoparticle-polymer composite materials can be optically printed into three-dimensional (3D) microstructures using digital projection printing. Piezoelectric polymers were fabricated by incorporating barium titanate (BaTiO3, BTO) nanoparticles into photoliable polymer solutions such as polyethylene glycol diacrylate and exposing to digital optical masks that could be dynamically altered to generate user-defined 3D microstructures. To enhance the mechanical-to-electrical conversion efficiency of the composites, the BTO nanoparticles were chemically modified with acrylate surface groups, which formed direct covalent linkages with the polymer matrix under light exposure. The composites with a 10% mass loading of the chemically modified BTO nanoparticles showed piezoelectric coefficients (d(33)) of ∼ 40 pC/N, which were over 10 times larger than composites synthesized with unmodified BTO nanoparticles and over 2 times larger than composites containing unmodified BTO nanoparticles and carbon nanotubes to boost mechanical stress transfer efficiencies. These results not only provide a tool for fabricating 3D piezoelectric polymers but lay the groundwork for creating highly efficient piezoelectric polymer materials via nanointerfacial tuning.

  8. Changes in quantitative 3D shape features of the optic nerve head associated with age

    NASA Astrophysics Data System (ADS)

    Christopher, Mark; Tang, Li; Fingert, John H.; Scheetz, Todd E.; Abramoff, Michael D.

    2013-02-01

    Optic nerve head (ONH) structure is an important biological feature of the eye used by clinicians to diagnose and monitor progression of diseases such as glaucoma. ONH structure is commonly examined using stereo fundus imaging or optical coherence tomography. Stereo fundus imaging provides stereo views of the ONH that retain 3D information useful for characterizing structure. In order to quantify 3D ONH structure, we applied a stereo correspondence algorithm to a set of stereo fundus images. Using these quantitative 3D ONH structure measurements, eigen structures were derived using principal component analysis from stereo images of 565 subjects from the Ocular Hypertension Treatment Study (OHTS). To evaluate the usefulness of the eigen structures, we explored associations with the demographic variables age, gender, and race. Using regression analysis, the eigen structures were found to have significant (p < 0.05) associations with both age and race after Bonferroni correction. In addition, classifiers were constructed to predict the demographic variables based solely on the eigen structures. These classifiers achieved an area under receiver operating characteristic curve of 0.62 in predicting a binary age variable, 0.52 in predicting gender, and 0.67 in predicting race. The use of objective, quantitative features or eigen structures can reveal hidden relationships between ONH structure and demographics. The use of these features could similarly allow specific aspects of ONH structure to be isolated and associated with the diagnosis of glaucoma, disease progression and outcomes, and genetic factors.

  9. 3D super-resolved in vitro multiphoton microscopy by saturation of excitation.

    PubMed

    Nguyen, Anh Dung; Duport, François; Bouwens, Arno; Vanholsbeeck, Frédérique; Egrise, Dominique; Van Simaeys, Gaetan; Emplit, Philippe; Goldman, Serge; Gorza, Simon-Pierre

    2015-08-24

    We demonstrate a significant resolution enhancement beyond the conventional limit in multiphoton microscopy (MPM) using saturated excitation of fluorescence. Our technique achieves super-resolved imaging by temporally modulating the excitation laser-intensity and demodulating the higher harmonics from the saturated fluorescence signal. The improvement of the lateral and axial resolutions is measured on a sample of fluorescent microspheres. While the third harmonic already provides an enhanced resolution, we show that a further improvement can be obtained with an appropriate linear combination of the demodulated harmonics. Finally, we present in vitro imaging of fluorescent microspheres incorporated in HeLa cells to show that this technique performs well in biological samples. PMID:26368235

  10. An enteroendocrine cell-enteric glia connection revealed by 3D electron microscopy.

    PubMed

    Bohórquez, Diego V; Samsa, Leigh A; Roholt, Andrew; Medicetty, Satish; Chandra, Rashmi; Liddle, Rodger A

    2014-01-01

    The enteroendocrine cell is the cornerstone of gastrointestinal chemosensation. In the intestine and colon, this cell is stimulated by nutrients, tastants that elicit the perception of flavor, and bacterial by-products; and in response, the cell secretes hormones like cholecystokinin and peptide YY--both potent regulators of appetite. The development of transgenic mice with enteroendocrine cells expressing green fluorescent protein has allowed for the elucidation of the apical nutrient sensing mechanisms of the cell. However, the basal secretory aspects of the enteroendocrine cell remain largely unexplored, particularly because a complete account of the enteroendocrine cell ultrastructure does not exist. Today, the fine ultrastructure of a specific cell can be revealed in the third dimension thanks to the invention of serial block face scanning electron microscopy (SBEM). Here, we bridged confocal microscopy with SBEM to identify the enteroendocrine cell of the mouse and study its ultrastructure in the third dimension. The results demonstrated that 73.5% of the peptide-secreting vesicles in the enteroendocrine cell are contained within an axon-like basal process. We called this process a neuropod. This neuropod contains neurofilaments, which are typical structural proteins of axons. Surprisingly, the SBEM data also demonstrated that the enteroendocrine cell neuropod is escorted by enteric glia--the cells that nurture enteric neurons. We extended these structural findings into an in vitro intestinal organoid system, in which the addition of glial derived neurotrophic factors enhanced the development of neuropods in enteroendocrine cells. These findings open a new avenue of exploration in gastrointestinal chemosensation by unveiling an unforeseen physical relationship between enteric glia and enteroendocrine cells. PMID:24587096

  11. An Enteroendocrine Cell – Enteric Glia Connection Revealed by 3D Electron Microscopy

    PubMed Central

    Bohórquez, Diego V.; Samsa, Leigh A.; Roholt, Andrew; Medicetty, Satish; Chandra, Rashmi; Liddle, Rodger A.

    2014-01-01

    The enteroendocrine cell is the cornerstone of gastrointestinal chemosensation. In the intestine and colon, this cell is stimulated by nutrients, tastants that elicit the perception of flavor, and bacterial by-products; and in response, the cell secretes hormones like cholecystokinin and peptide YY – both potent regulators of appetite. The development of transgenic mice with enteroendocrine cells expressing green fluorescent protein has allowed for the elucidation of the apical nutrient sensing mechanisms of the cell. However, the basal secretory aspects of the enteroendocrine cell remain largely unexplored, particularly because a complete account of the enteroendocrine cell ultrastructure does not exist. Today, the fine ultrastructure of a specific cell can be revealed in the third dimension thanks to the invention of serial block face scanning electron microscopy (SBEM). Here, we bridged confocal microscopy with SBEM to identify the enteroendocrine cell of the mouse and study its ultrastructure in the third dimension. The results demonstrated that 73.5% of the peptide-secreting vesicles in the enteroendocrine cell are contained within an axon-like basal process. We called this process a neuropod. This neuropod contains neurofilaments, which are typical structural proteins of axons. Surprisingly, the SBEM data also demonstrated that the enteroendocrine cell neuropod is escorted by enteric glia – the cells that nurture enteric neurons. We extended these structural findings into an in vitro intestinal organoid system, in which the addition of glial derived neurotrophic factors enhanced the development of neuropods in enteroendocrine cells. These findings open a new avenue of exploration in gastrointestinal chemosensation by unveiling an unforeseen physical relationship between enteric glia and enteroendocrine cells. PMID:24587096

  12. Highlighting the impact of aging on type I collagen: label-free investigation using confocal reflectance microscopy and diffuse reflectance spectroscopy in 3D matrix model.

    PubMed

    Guilbert, Marie; Roig, Blandine; Terryn, Christine; Garnotel, Roselyne; Jeannesson, Pierre; Sockalingum, Ganesh D; Manfait, Michel; Perraut, François; Dinten, Jean-Marc; Koenig, Anne; Piot, Olivier

    2016-02-23

    During aging, alterations of extracellular matrix proteins contribute to various pathological phenotypes. Among these alterations, type I collagen cross-linking and associated glycation products accumulation over time detrimentally affects its physico-chemical properties, leading to alterations of tissue biomechanical stability. Here, different-age collagen 3D matrices using non-destructive and label-free biophotonic techniques were analysed to highlight the impact of collagen I aging on 3D constructs, at macroscopic and microscopic levels. Matrices were prepared with collagens extracted from tail tendons of rats (newborns, young and old adults) to be within the physiological aging process. The data of diffuse reflectance spectroscopy reveal that aging leads to an inhibition of fibril assembly and a resulting decrease of gel density. Investigations by confocal reflectance microscopy highlight poor-fibrillar structures in oldest collagen networks most likely related to the glycation products accumulation. Complementarily, an infrared analysis brings out marked spectral variations in the Amide I profile, specific of the peptidic bond conformation and for carbohydrates vibrations as function of collagen-age. Interestingly, we also highlight an unexpected behavior for newborn collagen, exhibiting poorly-organized networks and microscopic features close to the oldest collagen. These results demonstrate that changes in collagen optical properties are relevant for investigating the incidence of aging in 3D matrix models.

  13. Highlighting the impact of aging on type I collagen: label-free investigation using confocal reflectance microscopy and diffuse reflectance spectroscopy in 3D matrix model

    PubMed Central

    Terryn, Christine; Garnotel, Roselyne; Jeannesson, Pierre; Sockalingum, Ganesh D.; Manfait, Michel; Perraut, François; Dinten, Jean-Marc; Koenig, Anne; Piot, Olivier

    2016-01-01

    During aging, alterations of extracellular matrix proteins contribute to various pathological phenotypes. Among these alterations, type I collagen cross-linking and associated glycation products accumulation over time detrimentally affects its physico-chemical properties, leading to alterations of tissue biomechanical stability. Here, different-age collagen 3D matrices using non-destructive and label-free biophotonic techniques were analysed to highlight the impact of collagen I aging on 3D constructs, at macroscopic and microscopic levels. Matrices were prepared with collagens extracted from tail tendons of rats (newborns, young and old adults) to be within the physiological aging process. The data of diffuse reflectance spectroscopy reveal that aging leads to an inhibition of fibril assembly and a resulting decrease of gel density. Investigations by confocal reflectance microscopy highlight poor-fibrillar structures in oldest collagen networks most likely related to the glycation products accumulation. Complementarily, an infrared analysis brings out marked spectral variations in the Amide I profile, specific of the peptidic bond conformation and for carbohydrates vibrations as function of collagen-age. Interestingly, we also highlight an unexpected behavior for newborn collagen, exhibiting poorly-organized networks and microscopic features close to the oldest collagen. These results demonstrate that changes in collagen optical properties are relevant for investigating the incidence of aging in 3D matrix models. PMID:26885896

  14. Measuring surface topography with scanning electron microscopy. I. EZEImage: a program to obtain 3D surface data.

    PubMed

    Ponz, Ezequiel; Ladaga, Juan Luis; Bonetto, Rita Dominga

    2006-04-01

    Scanning electron microscopy (SEM) is widely used in the science of materials and different parameters were developed to characterize the surface roughness. In a previous work, we studied the surface topography with fractal dimension at low scale and two parameters at high scale by using the variogram, that is, variance vs. step log-log graph, of a SEM image. Those studies were carried out with the FERImage program, previously developed by us. To verify the previously accepted hypothesis by working with only an image, it is indispensable to have reliable three-dimensional (3D) surface data. In this work, a new program (EZEImage) to characterize 3D surface topography in SEM has been developed. It uses fast cross correlation and dynamic programming to obtain reliable dense height maps in a few seconds which can be displayed as an image where each gray level represents a height value. This image can be used for the FERImage program or any other software to obtain surface topography characteristics. EZEImage also generates anaglyph images as well as characterizes 3D surface topography by means of a parameter set to describe amplitude properties and three functional indices for characterizing bearing and fluid properties. PMID:17481354

  15. GPU-based rapid reconstruction of cellular 3D refractive index maps from tomographic phase microscopy (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Dardikman, Gili; Shaked, Natan T.

    2016-03-01

    We present highly parallel and efficient algorithms for real-time reconstruction of the quantitative three-dimensional (3-D) refractive-index maps of biological cells without labeling, as obtained from the interferometric projections acquired by tomographic phase microscopy (TPM). The new algorithms are implemented on the graphic processing unit (GPU) of the computer using CUDA programming environment. The reconstruction process includes two main parts. First, we used parallel complex wave-front reconstruction of the TPM-based interferometric projections acquired at various angles. The complex wave front reconstructions are done on the GPU in parallel, while minimizing the calculation time of the Fourier transforms and phase unwrapping needed. Next, we implemented on the GPU in parallel the 3-D refractive index map retrieval using the TPM filtered-back projection algorithm. The incorporation of algorithms that are inherently parallel with a programming environment such as Nvidia's CUDA makes it possible to obtain real-time processing rate, and enables high-throughput platform for label-free, 3-D cell visualization and diagnosis.

  16. 3D map of the plant photosystem II supercomplex obtained by cryoelectron microscopy and single particle analysis.

    PubMed

    Nield, J; Orlova, E V; Morris, E P; Gowen, B; van Heel, M; Barber, J

    2000-01-01

    Here we describe the first 3D structure of the photosystem II (PSII) supercomplex of higher plants, constructed by single particle analysis of images obtained by cryoelectron microscopy. This large multisubunit membrane protein complex functions to absorb light energy and catalyze the oxidation of water and reduction of plastoquinone. The resolution of the 3D structure is 24 A and emphasizes the dimeric nature of the supercomplex. The extrinsic proteins of the oxygen-evolving complex (OEC) are readily observed as a tetrameric cluster bound to the lumenal surface. By considering higher resolution data, obtained from electron crystallography, it has been possible to relate the binding sites of the OEC proteins with the underlying intrinsic membrane subunits of the photochemical reaction center core. The model suggests that the 33 kDa OEC protein is located towards the CP47/D2 side of the reaction center but is also positioned over the C-terminal helices of the D1 protein including its CD lumenal loop. In contrast, the model predicts that the 23/17 kDa OEC proteins are positioned at the N-terminus of the D1 protein incorporating the AB lumenal loop of this protein and two other unidentified transmembrane helices. Overall the 3D model represents a significant step forward in revealing the structure of the photosynthetic OEC whose activity is required to sustain the aerobic atmosphere on our planet.

  17. Characterization of Polymer Blends: Optical Microscopy (*Polarized, Interference and Phase Contrast Microscopy*) and Confocal Microscopy

    SciTech Connect

    Ramanathan, Nathan Muruganathan; Darling, Seth B.

    2015-01-01

    Chapter 15 surveys the characterization of macro, micro and meso morphologies of polymer blends by optical microscopy. Confocal Microscopy offers the ability to view the three dimensional morphology of polymer blends, popular in characterization of biological systems. Confocal microscopy uses point illumination and a spatial pinhole to eliminate out-of focus light in samples that are thicker than the focal plane.

  18. Development of scanning laser sensor for underwater 3D imaging with the coaxial optics

    NASA Astrophysics Data System (ADS)

    Ochimizu, Hideaki; Imaki, Masaharu; Kameyama, Shumpei; Saito, Takashi; Ishibashi, Shoujirou; Yoshida, Hiroshi

    2014-06-01

    We have developed the scanning laser sensor for underwater 3-D imaging which has the wide scanning angle of 120º (Horizontal) x 30º (Vertical) with the compact size of 25 cm diameter and 60 cm long. Our system has a dome lens and a coaxial optics to realize both the wide scanning angle and the compactness. The system also has the feature in the sensitivity time control (STC) circuit, in which the receiving gain is increased according to the time of flight. The STC circuit contributes to detect a small signal by suppressing the unwanted signals backscattered by marine snows. We demonstrated the system performance in the pool, and confirmed the 3-D imaging with the distance of 20 m. Furthermore, the system was mounted on the autonomous underwater vehicle (AUV), and demonstrated the seafloor mapping at the depth of 100 m in the ocean.

  19. Optical absorption enhancement in 3D nanofibers coated on polymer substrate for photovoltaic devices.

    PubMed

    Kiani, Amirkianoosh; Venkatakrishnan, Krishnan; Tan, Bo

    2015-06-01

    Recent research in the field of photovoltaics has shown that polymer solar cells have great potential to provide low-cost, lightweight and flexible electronic devices to harvest solar energy. In this paper, we propose a new method for the generation of three-dimensional nanofibers coated on polymer substrate induced by femtosecond laser pulses. In this new method, a thin layer of polymer is irradiated by megahertz femtosecond laser pulses under ambient conditions, and a thin fibrous layer is generated on top of the polymer substrate. This method is single step; no additional materials are added, and the layers of the three-dimensional (3D) polymer nanofibrous structures are grown on top of the substrate after laser irradiation. Light spectroscopy results show significant enhancement of light absorption in the generated 3D nanofibrous layers of polymer. Finally, we suggest how to maximize the light trapping and optical absorption of the generated nanofiber cells by optimizing the laser parameters. PMID:26072881

  20. Seeing a Mycobacterium-Infected Cell in Nanoscale 3D: Correlative Imaging by Light Microscopy and FIB/SEM Tomography

    PubMed Central

    Beckwith, Marianne Sandvold; Beckwith, Kai Sandvold; Sikorski, Pawel; Skogaker, Nan Tostrup

    2015-01-01

    Mycobacteria pose a threat to the world health today, with pathogenic and opportunistic bacteria causing tuberculosis and non-tuberculous disease in large parts of the population. Much is still unknown about the interplay between bacteria and host during infection and disease, and more research is needed to meet the challenge of drug resistance and inefficient vaccines. This work establishes a reliable and reproducible method for performing correlative imaging of human macrophages infected with mycobacteria at an ultra-high resolution and in 3D. Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) tomography is applied, together with confocal fluorescence microscopy for localization of appropriately infected cells. The method is based on an Aclar poly(chloro-tri-fluoro)ethylene substrate, micropatterned into an advantageous geometry by a simple thermomoulding process. The platform increases the throughput and quality of FIB/SEM tomography analyses, and was successfully applied to detail the intracellular environment of a whole mycobacterium-infected macrophage in 3D. PMID:26406896

  1. Intrathoracic tumour motion estimation from CT imaging using the 3D optical flow method

    NASA Astrophysics Data System (ADS)

    Guerrero, Thomas; Zhang, Geoffrey; Huang, Tzung-Chi; Lin, Kang-Ping

    2004-09-01

    The purpose of this work was to develop and validate an automated method for intrathoracic tumour motion estimation from breath-hold computed tomography (BH CT) imaging using the three-dimensional optical flow method (3D OFM). A modified 3D OFM algorithm provided 3D displacement vectors for each voxel which were used to map tumour voxels on expiration BH CT onto inspiration BH CT images. A thoracic phantom and simulated expiration/inspiration BH CT pairs were used for validation. The 3D OFM was applied to the measured inspiration and expiration BH CT images from one lung cancer and one oesophageal cancer patient. The resulting displacements were plotted in histogram format and analysed to provide insight regarding the tumour motion. The phantom tumour displacement was measured as 1.20 and 2.40 cm with full-width at tenth maximum (FWTM) for the distribution of displacement estimates of 0.008 and 0.006 cm, respectively. The maximum error of any single voxel's motion estimate was 1.1 mm along the z-dimension or approximately one-third of the z-dimension voxel size. The simulated BH CT pairs revealed an rms error of less than 0.25 mm. The displacement of the oesophageal tumours was nonuniform and up to 1.4 cm, this was a new finding. A lung tumour maximum displacement of 2.4 cm was found in the case evaluated. In conclusion, 3D OFM provided an accurate estimation of intrathoracic tumour motion, with estimated errors less than the voxel dimension in a simulated motion phantom study. Surprisingly, oesophageal tumour motion was large and nonuniform, with greatest motion occurring at the gastro-oesophageal junction. Presented at The IASTED Second International Conference on Biomedical Engineering (BioMED 2004), Innsbruck, Austria, 16-18 February 2004.

  2. Three-Axis Distributed Fiber Optic Strain Measurement in 3D Woven Composite Structures

    NASA Technical Reports Server (NTRS)

    Castellucci, Matt; Klute, Sandra; Lally, Evan M.; Froggatt, Mark E.; Lowry, David

    2013-01-01

    Recent advancements in composite materials technologies have broken further from traditional designs and require advanced instrumentation and analysis capabilities. Success or failure is highly dependent on design analysis and manufacturing processes. By monitoring smart structures throughout manufacturing and service life, residual and operational stresses can be assessed and structural integrity maintained. Composite smart structures can be manufactured by integrating fiber optic sensors into existing composite materials processes such as ply layup, filament winding and three-dimensional weaving. In this work optical fiber was integrated into 3D woven composite parts at a commercial woven products manufacturing facility. The fiber was then used to monitor the structures during a VARTM manufacturing process, and subsequent static and dynamic testing. Low cost telecommunications-grade optical fiber acts as the sensor using a high resolution commercial Optical Frequency Domain Reflectometer (OFDR) system providing distributed strain measurement at spatial resolutions as low as 2mm. Strain measurements using the optical fiber sensors are correlated to resistive strain gage measurements during static structural loading. Keywords: fiber optic, distributed strain sensing, Rayleigh scatter, optical frequency domain reflectometry

  3. Analyzing Remodeling of Cardiac Tissue: A Comprehensive Approach Based on Confocal Microscopy and 3D Reconstructions.

    PubMed

    Seidel, Thomas; Edelmann, J-C; Sachse, Frank B

    2016-05-01

    Microstructural characterization of cardiac tissue and its remodeling in disease is a crucial step in many basic research projects. We present a comprehensive approach for three-dimensional characterization of cardiac tissue at the submicrometer scale. We developed a compression-free mounting method as well as labeling and imaging protocols that facilitate acquisition of three-dimensional image stacks with scanning confocal microscopy. We evaluated the approach with normal and infarcted ventricular tissue. We used the acquired image stacks for segmentation, quantitative analysis and visualization of important tissue components. In contrast to conventional mounting, compression-free mounting preserved cell shapes, capillary lumens and extracellular laminas. Furthermore, the new approach and imaging protocols resulted in high signal-to-noise ratios at depths up to 60 µm. This allowed extensive analyzes revealing major differences in volume fractions and distribution of cardiomyocytes, blood vessels, fibroblasts, myofibroblasts and extracellular space in control vs. infarct border zone. Our results show that the developed approach yields comprehensive data on microstructure of cardiac tissue and its remodeling in disease. In contrast to other approaches, it allows quantitative assessment of all major tissue components. Furthermore, we suggest that the approach will provide important data for physiological models of cardiac tissue at the submicrometer scale. PMID:26399990

  4. Analyzing Remodeling of Cardiac Tissue: A Comprehensive Approach Based on Confocal Microscopy and 3D Reconstructions.

    PubMed

    Seidel, Thomas; Edelmann, J-C; Sachse, Frank B

    2016-05-01

    Microstructural characterization of cardiac tissue and its remodeling in disease is a crucial step in many basic research projects. We present a comprehensive approach for three-dimensional characterization of cardiac tissue at the submicrometer scale. We developed a compression-free mounting method as well as labeling and imaging protocols that facilitate acquisition of three-dimensional image stacks with scanning confocal microscopy. We evaluated the approach with normal and infarcted ventricular tissue. We used the acquired image stacks for segmentation, quantitative analysis and visualization of important tissue components. In contrast to conventional mounting, compression-free mounting preserved cell shapes, capillary lumens and extracellular laminas. Furthermore, the new approach and imaging protocols resulted in high signal-to-noise ratios at depths up to 60 µm. This allowed extensive analyzes revealing major differences in volume fractions and distribution of cardiomyocytes, blood vessels, fibroblasts, myofibroblasts and extracellular space in control vs. infarct border zone. Our results show that the developed approach yields comprehensive data on microstructure of cardiac tissue and its remodeling in disease. In contrast to other approaches, it allows quantitative assessment of all major tissue components. Furthermore, we suggest that the approach will provide important data for physiological models of cardiac tissue at the submicrometer scale.

  5. Quantitative phase-amplitude microscopy I: optical microscopy.

    PubMed

    Barone-Nugent, E D; Barty, A; Nugent, K A

    2002-06-01

    In this paper, the application of a new optical microscopy method (quantitative phase-amplitude microscopy) to biological imaging is explored, and the issue of resolution and image quality is examined. The paper begins by presenting a theoretical analysis of the method using the optical transfer function formalism of Streibl (1985). The effect of coherence on the formation of the phase image is explored, and it is shown that the resolution of the method is not compromised over that of a conventional bright-field image. It is shown that the signal-to-noise ratio of the phase recovery, however, does depend on the degree of coherence in the illumination. Streibl (1985) notes that partially coherent image formation is a non-linear process because of the intermingling of amplitude and phase information. The work presented here shows that the quantitative phase-amplitude microscopy method acts to linearize the image formation process, and that the phase and amplitude information is properly described using a transfer function analysis. The theoretical conclusions are tested experimentally using an optical microscope and the theoretical deductions are confirmed. Samples for microscopy influence both the phase and amplitude of the light wave and it is demonstrated that the new phase recovery method can separate the amplitude and phase information, something not possible using traditional phase microscopy. In the case of a coherent wave, knowledge of the phase and amplitude constitutes complete information that can be used to emulate other forms of microscopy. This capacity is demonstrated by recovering the phase of a sample and using the data to emulate a differential interference contrast image.

  6. Focus Variation - A New Technology for High Resolution Optical 3D Surface Metrology

    NASA Astrophysics Data System (ADS)

    Scherer, S.

    2009-04-01

    Focus Variation - A New Technology for High Resolution Optical 3D Surface Metrology S. Scherer1, E. Cristea1, O. Huber1, A. Krenn1 1 ALICONA GmbH Graz, Austria The need for increasing accuracy is a characteristic of all geo-applications, and hence of the instruments contributing to obtaining relevant data. Small and fine sensors are being developed, measuring different parameters of our geosystem and requiring continuous validation and calibration. These sensors have often very small components (fine sensors able to sense dust, atmospheric water vapour characteristics, pressure change, gravimeters, satellite micro-components), showing complex topographies including steep flanks and having varying reflective properties. In order to get valid and reliable results, quality assurance of these instruments and sensors is required. The optical technology Focus-Variation, developed by Alicona and added in the latest draft of the upcoming ISO standard 25178, provides high resolution 3D surface metrology even at those complex topographies. The technique of Focus-Variation combines the small depth of focus of an optical system with vertical scanning to provide topographical and color information from the variation of focus. It is used for high-resolution optical 3D surface measurements. The traceable and repeatable measurement results are further being used for calibration and validation purposes. Some of the characteristics of the technology are: - Measurement of instruments / samples with steep flanks up to 80° - Measurement of materials with strongly varying reflection properties - Measurement of surfaces presenting fine (from 10nm) or strong roughness Here, we present the operating principle and possible applications of the optical 3D measurement system "InfiniteFocus", which is based on the technology of Focus-Variation and used for quality assurance in the lab and in production. With the vertical resolution of up to 10nm, InfiniteFocus yields meaningful form and

  7. Automated optical microscopy of coated particle fuel

    SciTech Connect

    Kercher, Andrew K; Hunn, John D; Price, Jeffery R; Pappano, Peter J

    2008-01-01

    Fundamental technological advances have occurred during the 20 year hiatus in US research on coated particle nuclear fuel. As part of the recent US Department of Energy s Advanced Gas Reactor Fuel Development and Qualification program, Oak Ridge National Laboratory has utilized advancements in computer automation, digital imaging, and image analysis to modernize US optical microscopy techniques for coated particle nuclear fuel. Automated optical microscopy has enabled detailed and objective analysis of individual particles (hundreds of measurements per particle) and of large sample sizes that far exceed the capabilities of conventional manual microscopy methods (analysis of 1500-5000 particles is common). Demonstrative examples of the capabilities of this automated optical microscopy are given for: (a) shadow imaging of kernels, coated fuel particles, and graphite matrix overcoated particles and (b) cross-sectional analysis of coated fuel particles to determine layer thicknesses.

  8. Extended volume and surface scatterometer for optical characterization of 3D-printed elements

    NASA Astrophysics Data System (ADS)

    Dannenberg, Florian; Uebeler, Denise; Weiß, Jürgen; Pescoller, Lukas; Weyer, Cornelia; Hahlweg, Cornelius

    2015-09-01

    The use of 3d printing technology seems to be a promising way for low cost prototyping, not only of mechanical, but also of optical components or systems. It is especially useful in applications where customized equipment repeatedly is subject to immediate destruction, as in experimental detonics and the like. Due to the nature of the 3D-printing process, there is a certain inner texture and therefore inhomogeneous optical behaviour to be taken into account, which also indicates mechanical anisotropy. Recent investigations are dedicated to quantification of optical properties of such printed bodies and derivation of corresponding optimization strategies for the printing process. Beside mounting, alignment and illumination means, also refractive and reflective elements are subject to investigation. The proposed measurement methods are based on an imaging nearfield scatterometer for combined volume and surface scatter measurements as proposed in previous papers. In continuation of last year's paper on the use of near field imaging, which basically is a reflective shadowgraph method, for characterization of glossy surfaces like printed matter or laminated material, further developments are discussed. The device has been extended for observation of photoelasticity effects and therefore homogeneity of polarization behaviour. A refined experimental set-up is introduced. Variation of plane of focus and incident angle are used for separation of various the images of the layers of the surface under test, cross and parallel polarization techniques are applied. Practical examples from current research studies are included.

  9. Fiber optic vibration sensor for high-power electric machines realized using 3D printing technology

    NASA Astrophysics Data System (ADS)

    Igrec, Bojan; Bosiljevac, Marko; Sipus, Zvonimir; Babic, Dubravko; Rudan, Smiljko

    2016-03-01

    The objective of this work was to demonstrate a lightweight and inexpensive fiber-optic vibration sensor, built using 3D printing technology, for high-power electric machines and similar applications. The working principle is based on modulating the light intensity using a blade attached to a bendable membrane. The sensor prototype was manufactured using PolyJet Matrix technology with DM 8515 Grey 35 Polymer. The sensor shows linear response, expected bandwidth (< 150 Hz), and from our measurements we estimated the damping ratio for used polymer to be ζ ≍ 0.019. The developed prototype is simple to assemble, adjust, calibrate and repair.

  10. Ultra-compact on-chip LED collimation optics by 3D femtosecond direct laser writing.

    PubMed

    Thiele, Simon; Gissibl, Timo; Giessen, Harald; Herkommer, Alois M

    2016-07-01

    By using two-photon lithographic 3D printing, we demonstrate additive manufacturing of a dielectric concentrator directly on a LED chip. With a size of below 200 μm in diameter and length, light output is increased by a factor of 6.2 in collimation direction, while the emission half-angle is reduced by 50%. We measure excellent form fidelity and irradiance patterns close to simulation. Additionally, a more complex shape design is presented, which exhibits a nonconventional triangular illumination pattern. The introduced method features exceptional design freedoms which can be used to tailor high-quality miniature illumination optics for specific lighting tasks, for example, endoscopy. PMID:27367093

  11. 3D imaging of translucent media with a plenoptic sensor based on phase space optics

    NASA Astrophysics Data System (ADS)

    Zhang, Xuanzhe; Shu, Bohong; Du, Shaojun

    2015-05-01

    Traditional stereo imaging technology is not working for dynamical translucent media, because there are no obvious characteristic patterns on it and it's not allowed using multi-cameras in most cases, while phase space optics can solve the problem, extracting depth information directly from "space-spatial frequency" distribution of the target obtained by plenoptic sensor with single lens. This paper discussed the presentation of depth information in phase space data, and calculating algorithms with different transparency. A 3D imaging example of waterfall was given at last.

  12. Influences of edges and steep slopes in 3D interference and confocal microscopy

    NASA Astrophysics Data System (ADS)

    Xie, Weichang; Hagemeier, Sebastian; Woidt, Carsten; Hillmer, Harmut; Lehmann, Peter

    2016-04-01

    Optical measurement techniques are widely applied in high-resolution contour, topography and roughness measurement. In this context vertical scanning white-light interferometers and confocal microscopes have become mature instruments over the last decades. The accuracy of measurement results is highly related not only to the type and physical properties of the measuring instruments, but also to the measurement object itself. This contribution focuses on measurement effects occurring at edges and height steps using white-light interferometers of different numerical apertures. If the edge is perfectly perpendicular, batwing effects appear at height steps. These batwings show maximum height if the height-to-wavelength-ratio (HWR) is about one forth or three forth, and they disappear if the HWR value is about an integer multiple of one half. The wavelength that is relevant in this context is the effective wavelength, i.e. the center wavelength of the illuminating light multiplied by a correction factor known as the numerical aperture correction. However, in practice the edges are usually not perfectly perpendicular. In this case, the measurement results depend also on the derivative of the surface height function and they may differ from theory and the prediction according to the HWR value. Measurements of such steps show systematical effects depending on the lateral resolution of the instrument. In this context, a Linnik interferometer with a magnification of 100x and NA = 0.9 is used to characterize the three dimensional topography of more or less rectangular calibration specimens and quasi-perpendicular structures produced by the nanoimprint technology. The Linnik interferometer is equipped with LED light sources emitting at different wavelengths, so that the HWR value can be changed. This is possible since the high NA objective lenses show a rather limited depth of focus such that the temporal coherence gating may be replaced by focal gating in this

  13. Influences of edges and steep slopes in 3D interference and confocal microscopy

    NASA Astrophysics Data System (ADS)

    Xie, Weichang; Hagemeier, Sebastian; Woidt, Carsten; Hillmer, Harmut; Lehmann, Peter

    2016-04-01

    Optical measurement techniques are widely applied in high-resolution contour, topography and roughness measurement. In this context vertical scanning white-light interferometers and confocal microscopes have become mature instruments over the last decades. The accuracy of measurement results is highly related not only to the type and physical properties of the measuring instruments, but also to the measurement object itself. This contribution focuses on measurement effects occurring at edges and height steps using white-light interferometers of different numerical apertures. If the edge is perfectly perpendicular, batwing effects appear at height steps. These batwings show maximum height if the height-to-wavelength-ratio (HWR) is about one forth or three forth, and they disappear if the HWR value is about an integer multiple of one half. The wavelength that is relevant in this context is the effective wavelength, i.e. the center wavelength of the illuminating light multiplied by a correction factor known as the numerical aperture correction. However, in practice the edges are usually not perfectly perpendicular. In this case, the measurement results depend also on the derivative of the surface height function and they may differ from theory and the prediction according to the HWR value. Measurements of such steps show systematical effects depending on the lateral resolution of the instrument. In this context, a Linnik interferometer with a magnification of 100x and NA = 0.9 is used to characterize the three dimensional topography of more or less rectangular calibration specimens and quasi-perpendicular structures produced by the nanoimprint technology. The Linnik interferometer is equipped with LED light sources emitting at different wavelengths, so that the HWR value can be changed. This is possible since the high NA objective lenses show a rather limited depth of focus such that the temporal coherence gating may be replaced by focal gating in this particular

  14. 3D optical sectioning with a new hyperspectral confocal fluorescence imaging system.

    SciTech Connect

    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.

  15. Polymer optical fibers integrated directly into 3D orthogonal woven composites for sensing

    NASA Astrophysics Data System (ADS)

    Hamouda, Tamer; Seyam, Abdel-Fattah M.; Peters, Kara

    2015-02-01

    This study demonstrates that standard polymer optical fibers (POF) can be directly integrated into composites from 3D orthogonal woven preforms during the weaving process and then serve as in-situ sensors to detect damage due to bending or impact loads. Different composite samples with embedded POF were fabricated of 3D orthogonal woven composites with different parameters namely number of y-/x-layers and x-yarn density. The signal of POF was not affected significantly by the preform structure. During application of resin using VARTM technique, significant drop in backscattering level was observed due to pressure caused by vacuum on the embedded POF. Measurements of POF signal while in the final composites after resin cure indicated that the backscattering level almost returned to the original level of un-embedded POF. The POF responded to application of bending and impact loads to the composite with a reduction in the backscattering level. The backscattering level almost returned back to its original level after removing the bending load until damage was present in the composite. Similar behavior occurred due to impact events. As the POF itself is used as the sensor and can be integrated throughout the composite, large sections of future 3D woven composite structures could be monitored without the need for specialized sensors or complex instrumentation.

  16. Diffractive 3D XUV optics at Helmholtz-Zentrum Berlin, recent developments

    NASA Astrophysics Data System (ADS)

    Brzhezinskaya, Maria; Firsov, Alexander; Erko, Alexei

    2014-09-01

    The 2-Dimensional and 3-Dimensional variable line spacing (VLS) gratings based on total external reflection give the unique possibility for spectroscopy and focusing in application to 4th and 5th generation synchrotron sources. We focus on the elaboration of novel approaches for design and fabrication of 3D VLS working in the entire energy range, from THz to hard X-rays. These optical elements have unique combination of properties and can operate at all XUV sources including Free Electron Lasers (FELs), Energy Recovery Linacs (ERLs) and High Harmonic Generators (HHGs). Such 3D DOEs are able to cover the energy range of up to 20 keV with energy resolution λ/Δλ ≥ 1000 for soft x-ray and λ/Δλ ≥ 10000 for hard x-ray. We fabricate 3D VLS for time-resolved spectroscopy (energy range 100 - 2000 eV, 7500-9500 eV), FELs and ERLs (energy range up to 3 keV), and HHGs (energy range 10 - 200 eV).

  17. Kinetic depth effect and optic flow--I. 3D shape from Fourier motion.

    PubMed

    Dosher, B A; Landy, M S; Sperling, G

    1989-01-01

    Fifty-three different 3D shapes were defined by sequences of 2D views (frames) of dots on a rotating 3D surface. (1) Subjects' accuracy of shape identifications dropped from over 90% to less than 10% when either the polarity of the stimulus dots was alternated from light-on-gray to dark-on-gray on successive frames or when neutral gray interframe intervals were interposed. Both manipulations interfere with motion extraction by spatio-temporal (Fourier) and gradient first-order detectors. Second-order (non-Fourier) detectors that use full-wave rectification are unaffected by alternating-polarity but disrupted by interposed gray frames. (2) To equate the accuracy of two-alternative forced-choice (2AFC) planar direction-of-motion discrimination in standard and polarity-alternated stimuli, standard contrast was reduced. 3D shape discrimination survived contrast reduction in standard stimuli whereas it failed completely with polarity-alternation even at full contrast. (3) When individual dots were permitted to remain in the image sequence for only two frames, performance showed little loss compared to standard displays where individual dots had an expected lifetime of 20 frames, showing that 3D shape identification does not require continuity of stimulus tokens. (4) Performance in all discrimination tasks is predicted (up to a monotone transformation) by considering the quality of first-order information (as given by a simple computation on Fourier power) and the number of locations at which motion information is required. Perceptual first-order analysis of optic flow is the primary substrate for structure-from-motion computations in random dot displays because only it offers sufficient quality of perceptual motion at a sufficient number of locations.

  18. Three-axis distributed fiber optic strain measurement in 3D woven composite structures

    NASA Astrophysics Data System (ADS)

    Castellucci, Matt; Klute, Sandra; Lally, Evan M.; Froggatt, Mark E.; Lowry, David

    2013-03-01

    Recent advancements in composite materials technologies have broken further from traditional designs and require advanced instrumentation and analysis capabilities. Success or failure is highly dependent on design analysis and manufacturing processes. By monitoring smart structures throughout manufacturing and service life, residual and operational stresses can be assessed and structural integrity maintained. Composite smart structures can be manufactured by integrating fiber optic sensors into existing composite materials processes such as ply layup, filament winding and three-dimensional weaving. In this work optical fiber was integrated into 3D woven composite parts at a commercial woven products manufacturing facility. The fiber was then used to monitor the structures during a VARTM manufacturing process, and subsequent static and dynamic testing. Low cost telecommunications-grade optical fiber acts as the sensor using a high resolution commercial Optical Frequency Domain Reflectometer (OFDR) system providing distributed strain measurement at spatial resolutions as low as 2mm. Strain measurements using the optical fiber sensors are correlated to resistive strain gage measurements during static structural loading.

  19. Combining supine MRI and 3D optical scanning for improved surgical planning of breast conserving surgeries

    NASA Astrophysics Data System (ADS)

    Pallone, Matthew J.; Poplack, Steven P.; Barth, Richard J., Jr.; Paulsen, Keith D.

    2012-02-01

    Image-guided wire localization is the current standard of care for the excision of non-palpable carcinomas during breast conserving surgeries (BCS). The efficacy of this technique depends upon the accuracy of wire placement, maintenance of the fixed wire position (despite patient movement), and the surgeon's understanding of the spatial relationship between the wire and tumor. Notably, breast shape can vary significantly between the imaging and surgical positions. Despite this method of localization, re-excision is needed in approximately 30% of patients due to the proximity of cancer to the specimen margins. These limitations make wire localization an inefficient and imprecise procedure. Alternatively, we investigate a method of image registration and finite element (FE) deformation which correlates preoperative supine MRIs with 3D optical scans of the breast surface. MRI of the breast can accurately define the extents of very small cancers. Furthermore, supine breast MR reduces the amount of tissue deformation between the imaging and surgical positions. At the time of surgery, the surface contour of the breast may be imaged using a handheld 3D laser scanner. With the MR images segmented by tissue type, the two scans are approximately registered using fiducial markers present in both acquisitions. The segmented MRI breast volume is then deformed to match the optical surface using a FE mechanical model of breast tissue. The resulting images provide the surgeon with 3D views and measurements of the tumor shape, volume, and position within the breast as it appears during surgery which may improve surgical guidance and obviate the need for wire localization.

  20. Optical-CT 3D Dosimetry Using Fresnel Lenses with Minimal Refractive-Index Matching Fluid

    PubMed Central

    Bache, Steven; Malcolm, Javian; Adamovics, John; Oldham, Mark

    2016-01-01

    Telecentric optical computed tomography (optical-CT) is a state-of-the-art method for visualizing and quantifying 3-dimensional dose distributions in radiochromic dosimeters. In this work a prototype telecentric system (DFOS—Duke Fresnel Optical-CT Scanner) is evaluated which incorporates two substantial design changes: the use of Fresnel lenses (reducing lens costs from $10-30K t0 $1-3K) and the use of a ‘solid tank’ (which reduces noise, and the volume of refractively matched fluid from 1ltr to 10cc). The efficacy of DFOS was evaluated by direct comparison against commissioned scanners in our lab. Measured dose distributions from all systems were compared against the predicted dose distributions from a commissioned treatment planning system (TPS). Three treatment plans were investigated including a simple four-field box treatment, a multiple small field delivery, and a complex IMRT treatment. Dosimeters were imaged within 2h post irradiation, using consistent scanning techniques (360 projections acquired at 1 degree intervals, reconstruction at 2mm). DFOS efficacy was evaluated through inspection of dose line-profiles, and 2D and 3D dose and gamma maps. DFOS/TPS gamma pass rates with 3%/3mm dose difference/distance-to-agreement criteria ranged from 89.3% to 92.2%, compared to from 95.6% to 99.0% obtained with the commissioned system. The 3D gamma pass rate between the commissioned system and DFOS was 98.2%. The typical noise rates in DFOS reconstructions were up to 3%, compared to under 2% for the commissioned system. In conclusion, while the introduction of a solid tank proved advantageous with regards to cost and convenience, further work is required to improve the image quality and dose reconstruction accuracy of the new DFOS optical-CT system. PMID:27019460

  1. Optical-CT 3D Dosimetry Using Fresnel Lenses with Minimal Refractive-Index Matching Fluid.

    PubMed

    Bache, Steven; Malcolm, Javian; Adamovics, John; Oldham, Mark

    2016-01-01

    Telecentric optical computed tomography (optical-CT) is a state-of-the-art method for visualizing and quantifying 3-dimensional dose distributions in radiochromic dosimeters. In this work a prototype telecentric system (DFOS-Duke Fresnel Optical-CT Scanner) is evaluated which incorporates two substantial design changes: the use of Fresnel lenses (reducing lens costs from $10-30K t0 $1-3K) and the use of a 'solid tank' (which reduces noise, and the volume of refractively matched fluid from 1 ltr to 10 cc). The efficacy of DFOS was evaluated by direct comparison against commissioned scanners in our lab. Measured dose distributions from all systems were compared against the predicted dose distributions from a commissioned treatment planning system (TPS). Three treatment plans were investigated including a simple four-field box treatment, a multiple small field delivery, and a complex IMRT treatment. Dosimeters were imaged within 2 h post irradiation, using consistent scanning techniques (360 projections acquired at 1 degree intervals, reconstruction at 2mm). DFOS efficacy was evaluated through inspection of dose line-profiles, and 2D and 3D dose and gamma maps. DFOS/TPS gamma pass rates with 3%/3mm dose difference/distance-to-agreement criteria ranged from 89.3% to 92.2%, compared to from 95.6% to 99.0% obtained with the commissioned system. The 3D gamma pass rate between the commissioned system and DFOS was 98.2%. The typical noise rates in DFOS reconstructions were up to 3%, compared to under 2% for the commissioned system. In conclusion, while the introduction of a solid tank proved advantageous with regards to cost and convenience, further work is required to improve the image quality and dose reconstruction accuracy of the new DFOS optical-CT system.

  2. Optical-CT 3D Dosimetry Using Fresnel Lenses with Minimal Refractive-Index Matching Fluid.

    PubMed

    Bache, Steven; Malcolm, Javian; Adamovics, John; Oldham, Mark

    2016-01-01

    Telecentric optical computed tomography (optical-CT) is a state-of-the-art method for visualizing and quantifying 3-dimensional dose distributions in radiochromic dosimeters. In this work a prototype telecentric system (DFOS-Duke Fresnel Optical-CT Scanner) is evaluated which incorporates two substantial design changes: the use of Fresnel lenses (reducing lens costs from $10-30K t0 $1-3K) and the use of a 'solid tank' (which reduces noise, and the volume of refractively matched fluid from 1 ltr to 10 cc). The efficacy of DFOS was evaluated by direct comparison against commissioned scanners in our lab. Measured dose distributions from all systems were compared against the predicted dose distributions from a commissioned treatment planning system (TPS). Three treatment plans were investigated including a simple four-field box treatment, a multiple small field delivery, and a complex IMRT treatment. Dosimeters were imaged within 2 h post irradiation, using consistent scanning techniques (360 projections acquired at 1 degree intervals, reconstruction at 2mm). DFOS efficacy was evaluated through inspection of dose line-profiles, and 2D and 3D dose and gamma maps. DFOS/TPS gamma pass rates with 3%/3mm dose difference/distance-to-agreement criteria ranged from 89.3% to 92.2%, compared to from 95.6% to 99.0% obtained with the commissioned system. The 3D gamma pass rate between the commissioned system and DFOS was 98.2%. The typical noise rates in DFOS reconstructions were up to 3%, compared to under 2% for the commissioned system. In conclusion, while the introduction of a solid tank proved advantageous with regards to cost and convenience, further work is required to improve the image quality and dose reconstruction accuracy of the new DFOS optical-CT system. PMID:27019460

  3. A correlative approach for combining microCT, light and transmission electron microscopy in a single 3D scenario

    PubMed Central

    2013-01-01

    Background In biomedical research, a huge variety of different techniques is currently available for the structural examination of small specimens, including conventional light microscopy (LM), transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), microscopic X-ray computed tomography (microCT), and many others. Since every imaging method is physically limited by certain parameters, a correlative use of complementary methods often yields a significant broader range of information. Here we demonstrate the advantages of the correlative use of microCT, light microscopy, and transmission electron microscopy for the analysis of small biological samples. Results We used a small juvenile bivalve mollusc (Mytilus galloprovincialis, approximately 0.8 mm length) to demonstrate the workflow of a correlative examination by microCT, LM serial section analysis, and TEM-re-sectioning. Initially these three datasets were analyzed separately, and subsequently they were fused in one 3D scene. This workflow is very straightforward. The specimen was processed as usual for transmission electron microscopy including post-fixation in osmium tetroxide and embedding in epoxy resin. Subsequently it was imaged with microCT. Post-fixation in osmium tetroxide yielded sufficient X-ray contrast for microCT imaging, since the X-ray absorption of epoxy resin is low. Thereafter, the same specimen was serially sectioned for LM investigation. The serial section images were aligned and specific organ systems were reconstructed based on manual segmentation and surface rendering. According to the region of interest (ROI), specific LM sections were detached from the slides, re-mounted on resin blocks and re-sectioned (ultrathin) for TEM. For analysis, image data from the three different modalities was co-registered into a single 3D scene using the software AMIRA®. We were able to register both the LM section series volume and TEM slices neatly to the microCT dataset, with

  4. Reconstruction and Visualization of Coordinated 3D Cell Migration Based on Optical Flow.

    PubMed

    Kappe, Christopher P; Schütz, Lucas; Gunther, Stefan; Hufnagel, Lars; Lemke, Steffen; Leitte, Heike

    2016-01-01

    Animal development is marked by the repeated reorganization of cells and cell populations, which ultimately determine form and shape of the growing organism. One of the central questions in developmental biology is to understand precisely how cells reorganize, as well as how and to what extent this reorganization is coordinated. While modern microscopes can record video data for every cell during animal development in 3D+t, analyzing these videos remains a major challenge: reconstruction of comprehensive cell tracks turned out to be very demanding especially with decreasing data quality and increasing cell densities. In this paper, we present an analysis pipeline for coordinated cellular motions in developing embryos based on the optical flow of a series of 3D images. We use numerical integration to reconstruct cellular long-term motions in the optical flow of the video, we take care of data validation, and we derive a LIC-based, dense flow visualization for the resulting pathlines. This approach allows us to handle low video quality such as noisy data or poorly separated cells, and it allows the biologists to get a comprehensive understanding of their data by capturing dynamic growth processes in stills. We validate our methods using three videos of growing fruit fly embryos.

  5. Coupling 3D Monte Carlo light transport in optically heterogeneous tissues to photoacoustic signal generation

    PubMed Central

    Jacques, Steven L.

    2014-01-01

    The generation of photoacoustic signals for imaging objects embedded within tissues is dependent on how well light can penetrate to and deposit energy within an optically absorbing object, such as a blood vessel. This report couples a 3D Monte Carlo simulation of light transport to stress wave generation to predict the acoustic signals received by a detector at the tissue surface. The Monte Carlo simulation allows modeling of optically heterogeneous tissues, and a simple MATLAB™ acoustic algorithm predicts signals reaching a surface detector. An example simulation considers a skin with a pigmented epidermis, a dermis with a background blood perfusion, and a 500-μm-dia. blood vessel centered at a 1-mm depth in the skin. The simulation yields acoustic signals received by a surface detector, which are generated by a pulsed 532-nm laser exposure before and after inserting the blood vessel. A MATLAB™ version of the acoustic algorithm and a link to the 3D Monte Carlo website are provided. PMID:25426426

  6. Coupling 3D Monte Carlo light transport in optically heterogeneous tissues to photoacoustic signal generation.

    PubMed

    Jacques, Steven L

    2014-12-01

    The generation of photoacoustic signals for imaging objects embedded within tissues is dependent on how well light can penetrate to and deposit energy within an optically absorbing object, such as a blood vessel. This report couples a 3D Monte Carlo simulation of light transport to stress wave generation to predict the acoustic signals received by a detector at the tissue surface. The Monte Carlo simulation allows modeling of optically heterogeneous tissues, and a simple MATLAB™ acoustic algorithm predicts signals reaching a surface detector. An example simulation considers a skin with a pigmented epidermis, a dermis with a background blood perfusion, and a 500-μm-dia. blood vessel centered at a 1-mm depth in the skin. The simulation yields acoustic signals received by a surface detector, which are generated by a pulsed 532-nm laser exposure before and after inserting the blood vessel. A MATLAB™ version of the acoustic algorithm and a link to the 3D Monte Carlo website are provided.

  7. Fast parallel interferometric 3D tracking of numerous optically trapped particles and their hydrodynamic interaction.

    PubMed

    Ruh, Dominic; Tränkle, Benjamin; Rohrbach, Alexander

    2011-10-24

    Multi-dimensional, correlated particle tracking is a key technology to reveal dynamic processes in living and synthetic soft matter systems. In this paper we present a new method for tracking micron-sized beads in parallel and in all three dimensions - faster and more precise than existing techniques. Using an acousto-optic deflector and two quadrant-photo-diodes, we can track numerous optically trapped beads at up to tens of kHz with a precision of a few nanometers by back-focal plane interferometry. By time-multiplexing the laser focus, we can calibrate individually all traps and all tracking signals in a few seconds and in 3D. We show 3D histograms and calibration constants for nine beads in a quadratic arrangement, although trapping and tracking is easily possible for more beads also in arbitrary 2D arrangements. As an application, we investigate the hydrodynamic coupling and diffusion anomalies of spheres trapped in a 3 × 3 arrangement. PMID:22109012

  8. How effective can optical-CT 3D dosimetry be without refractive fluid matching?

    PubMed Central

    Rankine, L; Oldham

    2013-01-01

    Achieving accurate optical CT 3D dosimetry without the use of viscous refractive index (RI) matching fluids would greatly increase convenience. Software has been developed to simulate optical CT 3D dosimetry for a range of scanning configurations including parallel-beam, point and converging light sources. For each configuration the efficacy of 3 refractive media were investigated: air, water, and a fluid closely matched to Presage (RI = 1.00, 1.33 and 1.49 respectively). The results revealed that the useable radius of the dosimeter (i.e. where data was within 2% of truth) reduced to 68% for water-matching, and 31% for dry-scanning in air. Point source incident ray geometry produced slightly more favourable results, although variation between the three geometries was relatively small. The required detector size however, increased by a factor six for dry-scanning, introducing cost penalties. For applications where dose information is not required in the periphery, some dry and low-viscous matching configurations may be feasible. PMID:24454520

  9. Improved Uav-Borne 3d Mapping by Fusing Optical and Laserscanner Data

    NASA Astrophysics Data System (ADS)

    Jutzi, B.; Weinmann, M.; Meidow, J.

    2013-08-01

    In this paper, a new method for fusing optical and laserscanner data is presented for improved UAV-borne 3D mapping. We propose to equip an unmanned aerial vehicle (UAV) with a small platform which includes two sensors: a standard low-cost digital camera and a lightweight Hokuyo UTM-30LX-EW laserscanning device (210 g without cable). Initially, a calibration is carried out for the utilized devices. This involves a geometric camera calibration and the estimation of the position and orientation offset between the two sensors by lever-arm and bore-sight calibration. Subsequently, a feature tracking is performed through the image sequence by considering extracted interest points as well as the projected 3D laser points. These 2D results are fused with the measured laser distances and fed into a bundle adjustment in order to obtain a Simultaneous Localization and Mapping (SLAM). It is demonstrated that an improvement in terms of precision for the pose estimation is derived by fusing optical and laserscanner data.

  10. High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization.

    PubMed

    Holden, Seamus J; Pengo, Thomas; Meibom, Karin L; Fernandez Fernandez, Carmen; Collier, Justine; Manley, Suliana

    2014-03-25

    We created a high-throughput modality of photoactivated localization microscopy (PALM) that enables automated 3D PALM imaging of hundreds of synchronized bacteria during all stages of the cell cycle. We used high-throughput PALM to investigate the nanoscale organization of the bacterial cell division protein FtsZ in live Caulobacter crescentus. We observed that FtsZ predominantly localizes as a patchy midcell band, and only rarely as a continuous ring, supporting a model of "Z-ring" organization whereby FtsZ protofilaments are randomly distributed within the band and interact only weakly. We found evidence for a previously unidentified period of rapid ring contraction in the final stages of the cell cycle. We also found that DNA damage resulted in production of high-density continuous Z-rings, which may obstruct cytokinesis. Our results provide a detailed quantitative picture of in vivo Z-ring organization.

  11. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

    DOE PAGES

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

    2016-05-01

    Here, we have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single moleculemore » super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.« less

  12. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution.

    PubMed

    Meddens, Marjolein B M; Liu, Sheng; Finnegan, Patrick S; Edwards, Thayne L; James, Conrad D; Lidke, Keith A

    2016-06-01

    We have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.

  13. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

    PubMed Central

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

    2016-01-01

    We have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet. PMID:27375939

  14. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

    SciTech Connect

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

    2016-01-01

    Here, we have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.

  15. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution.

    PubMed

    Meddens, Marjolein B M; Liu, Sheng; Finnegan, Patrick S; Edwards, Thayne L; James, Conrad D; Lidke, Keith A

    2016-06-01

    We have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet. PMID:27375939

  16. Direct Determination of 3D Distribution of Elemental Composition in Single Semiconductor Nanoislands by Scanning Auger Microscopy

    NASA Astrophysics Data System (ADS)

    Ponomaryov, Semyon S.; Yukhymchuk, Volodymyr O.; Lytvyn, Peter M.; Valakh, Mykhailo Ya

    2016-02-01

    An application of scanning Auger microscopy with ion etching technique and effective compensation of thermal drift of the surface analyzed area is proposed for direct local study of composition distribution in the bulk of single nanoislands. For GexSi1 - x-nanoislands obtained by MBE of Ge on Si-substrate gigantic interdiffusion mixing takes place both in the open and capped nanostructures. Lateral distributions of the elemental composition as well as concentration-depth profiles were recorded. 3D distribution of the elemental composition in the d-cluster bulk was obtained using the interpolation approach by lateral composition distributions in its several cross sections and concentration-depth profile. It was shown that there is a germanium core in the nanoislands of both nanostructure types, which even penetrates the substrate. In studied nanostructures maximal Ge content in the nanoislands may reach about 40 at.%.

  17. Sub-millimeter resolution 3D optical imaging of living tissue using laminar optical tomography

    PubMed Central

    Hillman, Elizabeth M. C.; Burgess, Sean A.

    2009-01-01

    In-vivo imaging of optical contrast in living tissues can allow measurement of functional parameters such as blood oxygenation and detection of targeted and active fluorescent contrast agents. However, optical imaging must overcome the effects of light scattering, which limit the penetration depth and can affect quantitation and sensitivity. This article focuses on a technique for high-resolution, high-speed depth-resolved optical imaging of superficial living tissues called laminar optical tomography (LOT), which is capable of imaging absorbing and fluorescent contrast in living tissues to depths of 2–3 mm with 100–200 micron resolution. An overview of the advantages and challenges of in-vivo optical imaging is followed by a review of currently available techniques for high-resolution optical imaging of tissues. LOT is then described, including a description of the imaging system design and discussion of data analysis and image reconstruction approaches. Examples of recent applications of LOT are then provided and compared to other existing technologies. By measuring multiply-scattered light, Laminar Optical Tomography can probe beneath the surface of living tissues such as the skin and brain. PMID:19844595

  18. Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI).

    PubMed

    Dertinger, T; Colyer, R; Iyer, G; Weiss, S; Enderlein, J

    2009-12-29

    Super-resolution optical microscopy is a rapidly evolving area of fluorescence microscopy with a tremendous potential for impacting many fields of science. Several super-resolution methods have been developed over the last decade, all capable of overcoming the fundamental diffraction limit of light. We present here an approach for obtaining subdiffraction limit optical resolution in all three dimensions. This method relies on higher-order statistical analysis of temporal fluctuations (caused by fluorescence blinking/intermittency) recorded in a sequence of images (movie). We demonstrate a 5-fold improvement in spatial resolution by using a conventional wide-field microscope. This resolution enhancement is achieved in iterative discrete steps, which in turn allows the evaluation of images at different resolution levels. Even at the lowest level of resolution enhancement, our method features significant background reduction and thus contrast enhancement and is demonstrated on quantum dot-labeled microtubules of fibroblast cells.

  19. Subwavelength optical microscopy in the far field

    SciTech Connect

    Sun Qingqing; Zubairy, M. Suhail; Al-Amri, M.; Scully, Marlan O.

    2011-06-15

    We present a procedure for subwavelength optical microscopy. The identical atoms are distributed on a plane and shined with a standing wave. We rotate the plane to different angles and record the resonant fluorescence spectra in the far field, from which we can obtain their distance and location information. This procedure also works for atomic separation above one wavelength and therefore provides a seamless microscopy.

  20. Optical 3D Nano-fabrication: Drawing or Growing? (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Kawata, Satoshi

    2016-05-01

    Conventional nanotechnology based on the lithography and scanning probe microscopy is limited to 2D fabrication and modification. Here, I will talk about the method for 3D laser fabrication with two-photon polymerization [1], two-photon isomerization [2], and two-photon photo-reduction [3]. Self-growth technology, such as self-grown fiber structures of polymer [4] and self-grown metallic fractal metamaterials structures [5] will be also discussed. [1] S. Kawata, et. al, Nature 412, 697-698, 2001. [2] S. Kawata and Y. Kawata, Chem Rev. 88, 083110, 2006. [3] Y. -Y. Cao, et. al., Small 5, 1144-1148, 2009 [4] S. Shoji and S. Kawata, Appl. Phys. Lett. 75, 737-739, 1999. [5] N. Takeyasu, N. Nishimura, S. Kawata, submitted.

  1. Optical Property Analyses of Plant Cells for Adaptive Optics Microscopy

    NASA Astrophysics Data System (ADS)

    Tamada, Yosuke; Murata, Takashi; Hattori, Masayuki; Oya, Shin; Hayano, Yutaka; Kamei, Yasuhiro; Hasebe, Mitsuyasu

    2014-04-01

    In astronomy, adaptive optics (AO) can be used to cancel aberrations caused by atmospheric turbulence and to perform diffraction-limited observation of astronomical objects from the ground. AO can also be applied to microscopy, to cancel aberrations caused by cellular structures and to perform high-resolution live imaging. As a step toward the application of AO to microscopy, here we analyzed the optical properties of plant cells. We used leaves of the moss Physcomitrella patens, which have a single layer of cells and are thus suitable for optical analysis. Observation of the cells with bright field and phase contrast microscopy, and image degradation analysis using fluorescent beads demonstrated that chloroplasts provide the main source of optical degradations. Unexpectedly, the cell wall, which was thought to be a major obstacle, has only a minor effect. Such information provides the basis for the application of AO to microscopy for the observation of plant cells.

  2. Development of portable 3D optical measuring system using structured light projection method

    NASA Astrophysics Data System (ADS)

    Aoki, Hiroshi

    2014-05-01

    Three-dimensional (3D) scanners are becoming increasingly common in many industries. However most of these scanning technologies have drawbacks for practical use due to size, weight, accessibility, and ease-of-use. Depending on the application, speed, flexibility and portability can often be deemed more important than accuracy. We have developed a solution to address this market requirement and overcome the aforementioned limitations. To counteract shortcomings such as heavy weight and large size, an optical sensor is used that consists of a laser projector, a camera system, and a multi-touch screen. Structured laser light is projected onto the measured object with a newly designed laser projector employing a single Micro Electro Mechanical Systems (MEMS) mirror. The optical system is optimized for the combination of a Laser Diode (LD), the MEMS mirror and the size of measurement area to secure the ideal contrast of structured light. Also, we developed a new calibration algorithm for this sensor with MEMS laser projector that uses an optical camera model for point cloud calculation. These technical advancements make the sensor compact, save power consumption, and reduce heat generation yet still allows for rapid calculation. Due to the principle of the measurement, structured light triangulation utilizing phase-shifting technology, resolution is improved. To meet requirements for practical applications, the optics, electronics, image processing, display and data management capabilities have been integrated into a single compact unit.

  3. Super-resolution optical microscopy: multiple choices.

    PubMed

    Huang, Bo

    2010-02-01

    The recent invention of super-resolution optical microscopy enables the visualization of fine features in biological samples with unprecedented clarity. It creates numerous opportunities in biology because vast amount of previously obscured subcellular processes now can be directly observed. Rapid development in this field in the past two years offers many imaging modalities that address different needs but they also complicates the choice of the 'perfect' method for answering a specific question. Here I will briefly describe the principles of super-resolution optical microscopy techniques and then focus on comparing their characteristics in various aspects of practical applications. PMID:19897404

  4. Direct observation of macrostructure formation of hierarchically structured meso-macroporous aluminosilicates with 3D interconnectivity by optical microscope.

    PubMed

    Lemaire, Arnaud; Rooke, Joanna Claire; Chen, Li-Hua; Su, Bao-Lian

    2011-03-15

    Hierarchically structured spongy meso-macroporous aluminosilicates with high tetrahedral aluminum content were synthesized from a mixture of single molecular alkoxide precursor, (sec-BuO)2-Al-O-Si(OEt)3, already containing Si-O-Al bonds, and a silica coreactant, tetramethoxysilane (TMOS). The spontaneous byproduct templated macroporous structure formation has been directly visualized using in situ high-resolution optical microscopy (OM), allowing the crucial observation of a microbubble dispersion which is directly correlated to the macrostructure observed by electronic microscopies (SEM and TEM). This discovery leads to a comparative study with meso-macroporous pure metal oxide and to a proposal of the formation mechanism of meso-macroporous aluminosilicates with 3D interconnectivity. The aluminosilicate phase/microbubbles emulsion is produced by a phase separation process occurring between the aluminosilicate nanoparticles and the liquid hydrolysis-condensation reaction byproducts (water, methanol, ethanol, and butanol). The use of alkoxysilane improves the heterocondensation rates between the highly reactive aluminum alkoxide part of the single precursor and added silica species but, above all, leads to the spontaneous generation of an unusual meso-macroporosity in alkaline media. The particles obtained at pH = 13.0 featured regular micrometer-sized macrospheres separated by very thin mesoporous walls and connected by submicrometric openings, providing a 3D interconnectivity. The slight increase in pH value to 13.5 induced significant modifications in morphology and textural properties due to the slower gelification process of the aluminosilicate phase, resulting in the formation of an aluminosilicate material constituted of 1-2 µm large independent hollow mesoporous spheres.

  5. Scanning all-fiber-optic endomicroscopy system for 3D nonlinear optical imaging of biological tissues

    PubMed Central

    Wu, Yicong; Leng, Yuxin; Xi, Jiefeng; Li, Xingde

    2009-01-01

    An extremely compact all-fiber-optic scanning endomicroscopy system was developed for two-photon fluorescence (TPF) and second harmonic generation (SHG) imaging of biological samples. A conventional double-clad fiber (DCF) was employed in the endomicroscope for single-mode femtosecond pulse delivery, multimode nonlinear optical signals collection and fast two-dimensional scanning. A single photonic bandgap fiber (PBF) with negative group velocity dispersion at two-photon excitation wavelength (i.e. ~810 nm) was used for pulse prechirping in replacement of a bulky grating/lens-based pulse stretcher. The combined use of DCF and PBF in the endomicroscopy system made the endomicroscope basically a plug-and-play unit. The excellent imaging ability of the extremely compact all-fiber-optic nonlinear optical endomicroscopy system was demonstrated by SHG imaging of rat tail tendon and depth-resolved TPF imaging of epithelial tissues stained with acridine orange. The preliminary results suggested the promising potential of this extremely compact all-fiber-optic endomicroscopy system for real-time assessment of both epithelial and stromal structures in luminal organs. PMID:19434122

  6. Full-color structured illumination optical sectioning microscopy.

    PubMed

    Qian, Jia; Lei, Ming; Dan, Dan; Yao, Baoli; Zhou, Xing; Yang, Yanlong; Yan, Shaohui; Min, Junwei; Yu, Xianghua

    2015-09-29

    In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology.

  7. Full-color structured illumination optical sectioning microscopy.

    PubMed

    Qian, Jia; Lei, Ming; Dan, Dan; Yao, Baoli; Zhou, Xing; Yang, Yanlong; Yan, Shaohui; Min, Junwei; Yu, Xianghua

    2015-01-01

    In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology. PMID:26415516

  8. Full-color structured illumination optical sectioning microscopy

    NASA Astrophysics Data System (ADS)

    Qian, Jia; Lei, Ming; Dan, Dan; Yao, Baoli; Zhou, Xing; Yang, Yanlong; Yan, Shaohui; Min, Junwei; Yu, Xianghua

    2015-09-01

    In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology.

  9. Full-color structured illumination optical sectioning microscopy

    PubMed Central

    Qian, Jia; Lei, Ming; Dan, Dan; Yao, Baoli; Zhou, Xing; Yang, Yanlong; Yan, Shaohui; Min, Junwei; Yu, Xianghua

    2015-01-01

    In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology. PMID:26415516

  10. Axial-Stereo 3-D Optical Metrology for Inner Profile of Pipes Using a Scanning Laser Endoscope

    PubMed Central

    Gong, Yuanzheng; Johnston, Richard S.; Melville, C. David; Seibel, Eric J.

    2015-01-01

    As the rapid progress in the development of optoelectronic components and computational power, 3D optical metrology becomes more and more popular in manufacturing and quality control due to its flexibility and high speed. However, most of the optical metrology methods are limited to external surfaces. This paper proposed a new approach to measure tiny internal 3D surfaces with a scanning fiber endoscope and axial-stereo vision algorithm. A dense, accurate point cloud of internally machined threads was generated to compare with its corresponding X-ray 3D data as ground truth, and the quantification was analyzed by Iterative Closest Points algorithm. PMID:26640425

  11. Axial-Stereo 3-D Optical Metrology for Inner Profile of Pipes Using a Scanning Laser Endoscope

    NASA Astrophysics Data System (ADS)

    Gong, Yuanzheng; Johnston, Richard S.; Melville, C. David; Seibel, Eric J.

    2015-07-01

    As the rapid progress in the development of optoelectronic components and computational power, 3-D optical metrology becomes more and more popular in manufacturing and quality control due to its flexibility and high speed. However, most of the optical metrology methods are limited to external surfaces. This article proposed a new approach to measure tiny internal 3-D surfaces with a scanning fiber endoscope and axial-stereo vision algorithm. A dense, accurate point cloud of internally machined threads was generated to compare with its corresponding X-ray 3-D data as ground truth, and the quantification was analyzed by Iterative Closest Points algorithm.

  12. Optical microscopy beyond the diffraction limit

    PubMed Central

    Smolyaninov, Igor I.

    2008-01-01

    Over the past century the resolution of far-field optical microscopes, which rely on propagating optical modes, was widely believed to be limited because of diffraction to a value on the order of a half-wavelength λ∕2 of the light used. Although immersion microscopes had slightly improved resolution on the order of λ∕2n, the increased resolution was limited by the small range of refractive indices, n, of available transparent materials. We are experiencing quick demolition of the diffraction limit in optical microscopy. Over the past few years numerous nonlinear optical microscopy techniques based on photoswitching and saturation of fluorescence demonstrated far-field resolution of 20 to 30 nm. The latest exciting example of these techniques has been demonstrated by Huang et al. [Science 319, 810–813 (2008)]. Moreover, recent progress in metamaterials indicates that artificial optical media can be created, which do not exhibit the diffraction limit. Resolution of linear “immersion” microscopes based on such metamaterials appears limited only by losses, which can be compensated by gain media. Thus, optical microscopy is quickly moving towards the 10 nm resolution scale, which should bring about numerous revolutionary advances in biomedical imaging. PMID:19404465

  13. 3D optical coherence tomography image registration for guiding cochlear implant insertion

    NASA Astrophysics Data System (ADS)

    Cheon, Gyeong-Woo; Jeong, Hyun-Woo; Chalasani, Preetham; Chien, Wade W.; Iordachita, Iulian; Taylor, Russell; Niparko, John; Kang, Jin U.

    2014-03-01

    In cochlear implant surgery, an electrode array is inserted into the cochlear canal to restore hearing to a person who is profoundly deaf or significantly hearing impaired. One critical part of the procedure is the insertion of the electrode array, which looks like a thin wire, into the cochlear canal. Although X-ray or computed tomography (CT) could be used as a reference to evaluate the pathway of the whole electrode array, there is no way to depict the intra-cochlear canal and basal turn intra-operatively to help guide insertion of the electrode array. Optical coherent tomography (OCT) is a highly effective way of visualizing internal structures of cochlea. Swept source OCT (SSOCT) having center wavelength of 1.3 micron and 2D Galvonometer mirrors was used to achieve 7-mm depth 3-D imaging. Graphics processing unit (GPU), OpenGL, C++ and C# were integrated for real-time volumetric rendering simultaneously. The 3D volume images taken by the OCT system were assembled and registered which could be used to guide a cochlear implant. We performed a feasibility study using both dry and wet temporal bones and the result is presented.

  14. Accuracy assessment of human trunk surface 3D reconstructions from an optical digitising system.

    PubMed

    Pazos, V; Cheriet, F; Song, L; Labelle, H; Dansereau, J

    2005-01-01

    The lack of reliable techniques to follow up scoliotic deformity from the external asymmetry of the trunk leads to a general use of X-rays and indices of spinal deformity. Young adolescents with idiopathic scoliosis need intensive follow-ups for many years and, consequently, they are repeatedly exposed to ionising radiation, which is hazardous to their long-term health. Furthermore, treatments attempt to improve both spinal and surface deformities, but internal indices do not describe the external asymmetry. The purpose of this study was to assess a commercial, optical 3D digitising system for the 3D reconstruction of the entire trunk for clinical assessment of external asymmetry. The resulting surface is a textured, high-density polygonal mesh. The accuracy assessment was based on repeated reconstructions of a manikin with markers fixed on it. The average normal distance between the reconstructed surfaces and the reference data (markers measured with CMM) was 1.1 +/- 0.9 mm. PMID:15742714

  15. Probing the 3D structure of cornea-like collagen liquid crystals with polarization-resolved SHG microscopy.

    PubMed

    Teulon, Claire; Tidu, Aurélien; Portier, François; Mosser, Gervaise; Schanne-Klein, Marie-Claire

    2016-07-11

    This work aims at characterizing the three-dimensional organization of liquid crystals composed of collagen, in order to determine the physico-chemical conditions leading to highly organized structures found in biological tissues such as cornea. To that end, we use second-harmonic generation (SHG) microscopy, since aligned collagen structures have been shown to exhibit intrinsic SHG signals. We combine polarization-resolved SHG experiments (P-SHG) with the theoretical derivation of the SHG signal of collagen molecules tilted with respect to the focal plane. Our P-SHG images exhibit striated patterns with variable contrast, as expected from our analytical and numerical calculations for plywood-like nematic structures similar to the ones found in the cornea. This study demonstrates the benefits of P-SHG microscopy for in situ characterization of highly organized biopolymers at micrometer scale, and the unique sensitivity of this nonlinear optical technique to the orientation of collagen molecules. PMID:27410876

  16. Particle-based optical pressure sensors for 3D pressure mapping.

    PubMed

    Banerjee, Niladri; Xie, Yan; Chalaseni, Sandeep; Mastrangelo, Carlos H

    2015-10-01

    This paper presents particle-based optical pressure sensors for in-flow pressure sensing, especially for microfluidic environments. Three generations of pressure sensitive particles have been developed- flat planar particles, particles with integrated retroreflectors and spherical microballoon particles. The first two versions suffer from pressure measurement dependence on particles orientation in 3D space and angle of interrogation. The third generation of microspherical particles with spherical symmetry solves these problems making particle-based manometry in microfluidic environment a viable and efficient methodology. Static and dynamic pressure measurements have been performed in liquid medium for long periods of time in a pressure range of atmospheric to 40 psi. Spherical particles with radius of 12 μm and balloon-wall thickness of 0.5 μm are effective for more than 5 h in this pressure range with an error of less than 5%.

  17. Full optical characterization of autostereoscopic 3D displays using local viewing angle and imaging measurements

    NASA Astrophysics Data System (ADS)

    Boher, Pierre; Leroux, Thierry; Bignon, Thibault; Collomb-Patton, Véronique

    2012-03-01

    Two commercial auto-stereoscopic 3D displays are characterized a using Fourier optics viewing angle system and an imaging video-luminance-meter. One display has a fixed emissive configuration and the other adapts its emission to the observer position using head tracking. For a fixed emissive condition, three viewing angle measurements are performed at three positions (center, right and left). Qualified monocular and binocular viewing spaces in front of the display are deduced as well as the best working distance. The imaging system is then positioned at this working distance and crosstalk homogeneity on the entire surface of the display is measured. We show that the crosstalk is generally not optimized on all the surface of the display. Display aspect simulation using viewing angle measurements allows understanding better the origin of those crosstalk variations. Local imperfections like scratches and marks generally increase drastically the crosstalk, demonstrating that cleanliness requirements for this type of display are quite critical.

  18. 3D optical phase reconstruction within PMMA samples using a spectral OCT system

    NASA Astrophysics Data System (ADS)

    Briones-R., Manuel d. J.; De La Torre-Ibarra, Manuel H.; Mendoza Santoyo, Fernando

    2015-08-01

    The optical coherence tomography (OCT) technique has proved to be a useful method in biomedical areas such as ophthalmology, dentistry, dermatology, among many others. In all these applications the main target is to reconstruct the internal structure of the samples from which the physician's expertise may recognize and diagnose the existence of a disease. Nowadays OCT has been applied one step further and is used to study the mechanics of some particular type of materials, where the resulting information involves more than just their internal structure and the measurement of parameters such as displacements, stress and strain. Here we report on a spectral OCT system used to image the internal 3D microstructure and displacement maps from a PMMA (Poly-methyl-methacrylate) sample, subjected to a deformation by a controlled three point bending and tilting. The internal mechanical response of the polymer is shown as consecutive 2D images.

  19. Coherent Nonlinear Optical Imaging: Beyond Fluorescence Microscopy

    PubMed Central

    Min, Wei; Freudiger, Christian W.; Lu, Sijia; Xie, X. Sunney

    2012-01-01

    The quest for ultrahigh detection sensitivity with spectroscopic contrasts other than fluorescence has led to various novel approaches to optical microscopy of biological systems. Coherent nonlinear optical imaging, especially the recently developed nonlinear dissipation microscopy, including stimulated Raman scattering and two photon absorption, and pump-probe microscopy, including stimulated emission, excited state absorption and ground state depletion, provide distinct and powerful image contrasts for non-fluorescent species. Thanks to high-frequency modulation transfer scheme, they exhibit superb detection sensitivity. By directly interrogating vibrational and/or electronic energy levels of molecules, they offer high molecular specificity. Here we review the underlying principles, excitation and detection schemes, as well as exemplary biomedical applications of this emerging class of molecular imaging techniques. PMID:21453061

  20. Intra-retinal layer segmentation of 3D optical coherence tomography using coarse grained diffusion map

    PubMed Central

    Kafieh, Raheleh; Rabbani, Hossein; Abramoff, Michael D.; Sonka, Milan

    2013-01-01

    Optical coherence tomography (OCT) is a powerful and noninvasive method for retinal imaging. In this paper, we introduce a fast segmentation method based on a new variant of spectral graph theory named diffusion maps. The research is performed on spectral domain (SD) OCT images depicting macular and optic nerve head appearance. The presented approach does not require edge-based image information in localizing most of boundaries and relies on regional image texture. Consequently, the proposed method demonstrates robustness in situations of low image contrast or poor layer-to-layer image gradients. Diffusion mapping applied to 2D and 3D OCT datasets is composed of two steps, one for partitioning the data into important and less important sections, and another one for localization of internal layers. In the first step, the pixels/voxels are grouped in rectangular/cubic sets to form a graph node. The weights of the graph are calculated based on geometric distances between pixels/voxels and differences of their mean intensity. The first diffusion map clusters the data into three parts, the second of which is the area of interest. The other two sections are eliminated from the remaining calculations. In the second step, the remaining area is subjected to another diffusion map assessment and the internal layers are localized based on their textural similarities. The proposed method was tested on 23 datasets from two patient groups (glaucoma and normals). The mean unsigned border positioning errors (mean ± SD) was 8.52 ± 3.13 and 7.56 ± 2.95 μm for the 2D and 3D methods, respectively. PMID:23837966

  1. Structure-function studies of blood and air capillaries in chicken lung using 3D electron microscopy.

    PubMed

    West, John B; Fu, Zhenxing; Deerinck, Thomas J; Mackey, Mason R; Obayashi, James T; Ellisman, Mark H

    2010-02-28

    Avian pulmonary capillaries differ from those of mammals in three important ways. The blood-gas barrier is much thinner, it is more uniform in thickness, and the capillaries are far more rigid when their transmural pressure is altered. The thinness of the barrier is surprising because it predisposes the capillaries to stress failure. A possible mechanism for these differences is that avian pulmonary capillaries, unlike mammalian, are supported from the outside by air capillaries, but the details of the support are poorly understood. To clarify this we studied the blood and air capillaries in chicken lung using transmission electron microscopy (EM) and two relatively new techniques that allow 3D visualization: electron tomography and serial block-face scanning EM. These studies show that the pulmonary capillaries are flanked by epithelial bridges composed of two extremely thin epithelial cells with large surface areas. The junctions of the bridges with the capillary walls show thickening of the epithelial cells and an accumulation of extracellular matrix. Collapse of the pulmonary capillaries when the pressure outside them is increased is apparently prevented by the guy wire-like action of the epithelial bridges. The enlarged junctions between the bridges and the walls could provide a mechanism that limits the hoop stress in the capillary walls when the pressure inside them is increased. The support of the pulmonary capillaries may also be explained by an interdependence mechanism whereby the capillaries are linked to a rigid assemblage of air capillaries. These EM studies show the supporting structures in greater detail than has previously been possible, particularly in 3D, and they allow a more complete analysis of the mechanical forces affecting avian pulmonary capillaries. PMID:20038456

  2. Rat Optic Nerve Head Anatomy within 3D Histomorphometric Reconstructions of Normal Control Eyes

    PubMed Central

    Pazos, Marta; Yang, Hongli; Gardiner, Stuart K.; Cepurna, William O.; Johnson, Elaine C.; Morrison, John C.; Burgoyne, Claude F.

    2015-01-01

    The purpose of this study is to three-dimensionally (3D) characterize the principal macroscopic and microscopic relationships within the rat optic nerve head (ONH) and quantify them in normal control eyes. Perfusion-fixed, trephinated ONH from 8 normal control eyes of 8 Brown Norway Rats were 3D histomorphometrically reconstructed, visualized, delineated and parameterized. The rat ONH consists of 2 scleral openings, (a superior neurovascular and inferior arterial) separated by a thin connective tissue strip we have termed the “scleral sling”. Within the superior opening, the nerve abuts a prominent extension of Bruch's Membrane (BM) superiorly and is surrounded by a vascular plexus, as it passes through the sclera, that is a continuous from the choroid into and through the dural sheath and contains the central retinal vein (CRV), (inferiorly). The inferior scleral opening contains the central retinal artery and three long posterior ciliary arteries which obliquely pass through the sclera to obtain the choroid. Bruch's Membrane Opening (BMO) is irregular and vertically elongated, enclosing the nerve (superiorly) and CRV and CRA (inferiorly). Overall mean BMO Depth, BMO Area, Choroidal Thickness and peripapillary Scleral Thickness were 29 μm, 56.5 × 103 μm2, 57 μm and 104 μm respectively. Mean anterior scleral canal opening (ASCO) and posterior scleral canal opening (PSCO) radii were 201 ± 15 μm and 204 ± 16 μm, respectively. Mean optic nerve area at the ASCO and PSCO were 46.3 × 103 ± 4.4 × 103 μm2 and 44.1 × 103 ± 4.5 × 103 μm2 respectively. In conclusion, the 3D complexity of the rat ONH and the extent to which it differs from the primate have been under-appreciated within previous 2D studies. Properly understood, these anatomic differences may provide new insights into the relative susceptibilities of the rat and primate ONH to elevated intraocular pressure. PMID:26021973

  3. Near-Field Scanning Optical Microscopy and Raman Microscopy.

    NASA Astrophysics Data System (ADS)

    Harootunian, Alec Tate

    1987-09-01

    Both a one dimensional near-field scanning optical microscope and Raman microprobe were constructed. In near -field scanning optical microscopy (NSOM) a subwavelength aperture is scanned in the near-field of the object. Radiation transmitted through the aperture is collected to form an image as the aperture scans over the object. The resolution of an NSOM system is essentially wavelength independent and is limited by the diameter of the aperture used to scan the object. NSOM was developed in an effort to provide a nondestructive in situ high spatial resolution probe while still utilizing photons at optical wavelengths. The Raman microprobe constructed provided vibrational Raman information with spatial resolution equivalent that of a conventional diffraction limited microscope. Both transmission studies and near-field diffration studies of subwavelength apertures were performed. Diffraction theories for a small aperture in an infinitely thin conducting screen, a slit in a thick conducting screen, and an aperture in a black screen were examined. All three theories indicate collimation of radiation to the size to the size of the subwavelength aperture or slit in the near-field. Theoretical calculations and experimental results indicate that light transmitted through subwavelength apertures is readily detectable. Light of wavelength 4579 (ANGSTROM) was transmitted through apertures with diameters as small as 300 (ANGSTROM). These studies indicate the feasibility of constructing an NSOM system. One dimensional transmission and fluorescence NSOM systems were constructed. Apertures in the tips of metallized glass pipettes width inner diameters of less than 1000 (ANGSTROM) were used as a light source in the NSOM system. A tunneling current was used to maintain the aperture position in the near-field. Fluorescence NSOM was demonstrated for the first time. Microspectroscopic and Raman microscopic studies of turtle cone oil droplets were performed. Both the Raman vibrational

  4. Beyond optical molasses: 3D raman sideband cooling of atomic cesium to high phase-space density

    PubMed

    Kerman; Vuletic; Chin; Chu

    2000-01-17

    We demonstrate a simple, general purpose method to cool neutral atoms. A sample containing 3x10(8) cesium atoms prepared in a magneto-optical trap is cooled and simultaneously spin polarized in 10 ms at a density of 1.1x10(11) cm (-3) to a phase space density nlambda(3)(dB) = 1/500, which is almost 3 orders of magnitude higher than attainable in free space with optical molasses. The technique is based on 3D degenerate Raman sideband cooling in optical lattices and remains efficient even at densities where the mean lattice site occupation is close to unity.

  5. 3-D laser confocal microscopy study of the oxidation of NdFeB magnets in atmospheric conditions

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

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

  6. Automatic registration of optical imagery with 3d lidar data using local combined mutual information

    NASA Astrophysics Data System (ADS)

    Parmehr, E. G.; Fraser, C. S.; Zhang, C.; Leach, J.

    2013-10-01

    Automatic registration of multi-sensor data is a basic step in data fusion for photogrammetric and remote sensing applications. The effectiveness of intensity-based methods such as Mutual Information (MI) for automated registration of multi-sensor image has been previously reported for medical and remote sensing applications. In this paper, a new multivariable MI approach that exploits complementary information of inherently registered LiDAR DSM and intensity data to improve the robustness of registering optical imagery and LiDAR point cloud, is presented. LiDAR DSM and intensity information has been utilised in measuring the similarity of LiDAR and optical imagery via the Combined MI. An effective histogramming technique is adopted to facilitate estimation of a 3D probability density function (pdf). In addition, a local similarity measure is introduced to decrease the complexity of optimisation at higher dimensions and computation cost. Therefore, the reliability of registration is improved due to the use of redundant observations of similarity. The performance of the proposed method for registration of satellite and aerial images with LiDAR data in urban and rural areas is experimentally evaluated and the results obtained are discussed.

  7. Label-free optical detection of cells grown in 3D silicon microstructures.

    PubMed

    Merlo, Sabina; Carpignano, Francesca; Silva, Gloria; Aredia, Francesca; Scovassi, A Ivana; Mazzini, Giuliano; Surdo, Salvatore; Barillaro, Giuseppe

    2013-08-21

    We demonstrate high aspect-ratio photonic crystals that could serve as three-dimensional (3D) microincubators for cell culture and also provide label-free optical detection of the cells. The investigated microstructures, fabricated by electrochemical micromachining of standard silicon wafers, consist of periodic arrays of silicon walls separated by narrow deeply etched air-gaps (50 μm high and 5 μm wide) and feature the typical spectral properties of photonic crystals in the wavelength range 1.0-1.7 μm: their spectral reflectivity is characterized by wavelength regions where reflectivity is high (photonic bandgaps), separated by narrow wavelength regions where reflectivity is very low. In this work, we show that the presence of cells, grown inside the gaps, strongly affects light propagation across the photonic crystal and, therefore, its spectral reflectivity. Exploiting a label-free optical detection method, based on a fiberoptic setup, we are able to probe the extension of cells adherent to the vertical silicon walls with a non-invasive direct testing. In particular, the intensity ratio at two wavelengths is the experimental parameter that can be well correlated to the cell spreading on the silicon wall inside the gaps.

  8. Lateral error reduction in the 3D characterization of deep MOEMS devices using white light interference microscopy

    NASA Astrophysics Data System (ADS)

    Montgomery, Paul C.; Montaner, Denis; Manzardo, Omar; Herzig, Hans-Peter

    2004-08-01

    White light scanning interference microscopy, with its high axial resolution, is particularly useful for the rapid 3D characterization of MOEMS micro-systems. Although this technique can be used for submicron critical dimension measurement on micron high microelectronic structures, recent tests using a standard system have revealed errors of up to 3 μm in the measurement of lateral position of deep square steps. Thus the 2 μm wide, 75 μm deep teeth of an electrostatic comb structure in a FT MOEMS spectrometer were measured to be nearly 7 μm wide using a Mirau interference objective with the aperture diaphragm of the illumination system fully open in white light. Tests under different conditions show that the error is greatest for the Mirau objective, with the aperture diaphragm fully open at longer wavelengths. In addition, the location of the centre of such structures can vary by up to 1 μm depending on the degree of reference mirror tilt. Investigations of the XZ images of square steps have revealed the presence of "ghost" fringes resulting from diffraction and the conical illumination used. The errors in edge position can be reduced using a Linnik type objective with the aperture diaphragm closed down using shorter wavelength light.

  9. A workflow to process 3D+time microscopy images of developing organisms and reconstruct their cell lineage.

    PubMed

    Faure, Emmanuel; Savy, Thierry; Rizzi, Barbara; Melani, Camilo; Stašová, Olga; Fabrèges, Dimitri; Špir, Róbert; Hammons, Mark; Čúnderlík, Róbert; Recher, Gaëlle; Lombardot, Benoît; Duloquin, Louise; Colin, Ingrid; Kollár, Jozef; Desnoulez, Sophie; Affaticati, Pierre; Maury, Benoît; Boyreau, Adeline; Nief, Jean-Yves; Calvat, Pascal; Vernier, Philippe; Frain, Monique; Lutfalla, Georges; Kergosien, Yannick; Suret, Pierre; Remešíková, Mariana; Doursat, René; Sarti, Alessandro; Mikula, Karol; Peyriéras, Nadine; Bourgine, Paul

    2016-01-01

    The quantitative and systematic analysis of embryonic cell dynamics from in vivo 3D+time image data sets is a major challenge at the forefront of developmental biology. Despite recent breakthroughs in the microscopy imaging of living systems, producing an accurate cell lineage tree for any developing organism remains a difficult task. We present here the BioEmergences workflow integrating all reconstruction steps from image acquisition and processing to the interactive visualization of reconstructed data. Original mathematical methods and algorithms underlie image filtering, nucleus centre detection, nucleus and membrane segmentation, and cell tracking. They are demonstrated on zebrafish, ascidian and sea urchin embryos with stained nuclei and membranes. Subsequent validation and annotations are carried out using Mov-IT, a custom-made graphical interface. Compared with eight other software tools, our workflow achieved the best lineage score. Delivered in standalone or web service mode, BioEmergences and Mov-IT offer a unique set of tools for in silico experimental embryology. PMID:26912388

  10. A workflow to process 3D+time microscopy images of developing organisms and reconstruct their cell lineage

    PubMed Central

    Faure, Emmanuel; Savy, Thierry; Rizzi, Barbara; Melani, Camilo; Stašová, Olga; Fabrèges, Dimitri; Špir, Róbert; Hammons, Mark; Čúnderlík, Róbert; Recher, Gaëlle; Lombardot, Benoît; Duloquin, Louise; Colin, Ingrid; Kollár, Jozef; Desnoulez, Sophie; Affaticati, Pierre; Maury, Benoît; Boyreau, Adeline; Nief, Jean-Yves; Calvat, Pascal; Vernier, Philippe; Frain, Monique; Lutfalla, Georges; Kergosien, Yannick; Suret, Pierre; Remešíková, Mariana; Doursat, René; Sarti, Alessandro; Mikula, Karol; Peyriéras, Nadine; Bourgine, Paul

    2016-01-01

    The quantitative and systematic analysis of embryonic cell dynamics from in vivo 3D+time image data sets is a major challenge at the forefront of developmental biology. Despite recent breakthroughs in the microscopy imaging of living systems, producing an accurate cell lineage tree for any developing organism remains a difficult task. We present here the BioEmergences workflow integrating all reconstruction steps from image acquisition and processing to the interactive visualization of reconstructed data. Original mathematical methods and algorithms underlie image filtering, nucleus centre detection, nucleus and membrane segmentation, and cell tracking. They are demonstrated on zebrafish, ascidian and sea urchin embryos with stained nuclei and membranes. Subsequent validation and annotations are carried out using Mov-IT, a custom-made graphical interface. Compared with eight other software tools, our workflow achieved the best lineage score. Delivered in standalone or web service mode, BioEmergences and Mov-IT offer a unique set of tools for in silico experimental embryology. PMID:26912388

  11. Correction of depth-dependent aberrations in 3D single-molecule localization and super-resolution microscopy.

    PubMed

    McGorty, Ryan; Schnitzbauer, Joerg; Zhang, Wei; Huang, Bo

    2014-01-15

    Single-molecule switching based super-resolution microscopy techniques have been extended into three dimensions through various 3D single-molecule localization methods. However, the localization accuracy in z can be severely degraded by the presence of aberrations, particularly the spherical aberration introduced by the refractive index mismatch when imaging into an aqueous sample with an oil immersion objective. This aberration confines the imaging depth in most experiments to regions close to the coverslip. Here we show a method to obtain accurate, depth-dependent z calibrations by measuring the point spread function (PSF) at the coverslip surface, calculating the microscope pupil function through phase retrieval, and then computing the depth-dependent PSF with the addition of spherical aberrations. We demonstrate experimentally that this method can maintain z localization accuracy over a large range of imaging depths. Our super-resolution images of a mammalian cell nucleus acquired between 0 and 2.5 μm past the coverslip show that this method produces accurate z localizations even in the deepest focal plane.

  12. A new high-aperture glycerol immersion objective lens and its application to 3D-fluorescence microscopy.

    PubMed

    Martini, N; Bewersdorf, J; Hell, S W

    2002-05-01

    High-resolution light microscopy of glycerol-mounted biological specimens is performed almost exclusively with oil immersion lenses. The reason is that the index of refraction of the oil and the cover slip of approximately 1.51 is close to that of approximately 1.45 of the glycerol mountant, so that refractive index mismatch-induced spherical aberrations are tolerable to some extent. Here we report the application of novel cover glass-corrected glycerol immersion lenses of high numerical aperture (NA) and the avoidance of these aberrations. The new lenses feature a semi-aperture angle of 68.5 degrees, which is slightly larger than that of the diffraction-limited 1.4 NA oil immersion lenses. The glycerol lenses are corrected for a quartz cover glass of 220 microm thickness and for a 80% glycerol-water immersion solution. Featuring an aberration correction collar, the lens can adapt to glycerol concentrations ranging between 72% and 88%, to slight variations of the temperature, and to the cover glass thickness. As the refractive index mismatch-induced aberrations are particularly important to quantitative confocal fluorescence microscopy, we investigated the axial sectioning ability and the axial chromatic aberrations in such a microscope as well as the image brightness as a function of the penetration depth. Whereas there is a significant decrease in image brightness associated with oil immersion, this decrease is absent with the glycerol immersion system. In addition, we show directly the compression of the optic axis in the case of oil immersion and its absence in the glycerol system. The unique advantages of these new lenses in high-resolution microscopy with two coherently used opposing lenses, such as 4 Pi-microscopy, are discussed. PMID:12000554

  13. Real-time 3D Fourier-domain optical coherence tomography guided microvascular anastomosis

    NASA Astrophysics Data System (ADS)

    Huang, Yong; Ibrahim, Zuhaib; Lee, W. P. A.; Brandacher, Gerald; Kang, Jin U.

    2013-03-01

    Vascular and microvascular anastomosis is considered to be the foundation of plastic and reconstructive surgery, hand surgery, transplant surgery, vascular surgery and cardiac surgery. In the last two decades innovative techniques, such as vascular coupling devices, thermo-reversible poloxamers and suture-less cuff have been introduced. Intra-operative surgical guidance using a surgical imaging modality that provides in-depth view and 3D imaging can improve outcome following both conventional and innovative anastomosis techniques. Optical coherence tomography (OCT) is a noninvasive high-resolution (micron level), high-speed, 3D imaging modality that has been adopted widely in biomedical and clinical applications. In this work we performed a proof-of-concept evaluation study of OCT as an assisted intraoperative and post-operative imaging modality for microvascular anastomosis of rodent femoral vessels. The OCT imaging modality provided lateral resolution of 12 μm and 3.0 μm axial resolution in air and 0.27 volume/s imaging speed, which could provide the surgeon with clearly visualized vessel lumen wall and suture needle position relative to the vessel during intraoperative imaging. Graphics processing unit (GPU) accelerated phase-resolved Doppler OCT (PRDOCT) imaging of the surgical site was performed as a post-operative evaluation of the anastomosed vessels and to visualize the blood flow and thrombus formation. This information could help surgeons improve surgical precision in this highly challenging anastomosis of rodent vessels with diameter less than 0.5 mm. Our imaging modality could not only detect accidental suture through the back wall of lumen but also promptly diagnose and predict thrombosis immediately after reperfusion. Hence, real-time OCT can assist in decision-making process intra-operatively and avoid post-operative complications.

  14. Nonlinear Optical Macroscopic Assessment of 3-D Corneal Collagen Organization and Axial Biomechanics

    PubMed Central

    Winkler, Moritz; Chai, Dongyul; Kriling, Shelsea; Nien, Chyong Jy; Brown, Donald J.; Jester, Bryan; Juhasz, Tibor

    2011-01-01

    Purpose. To characterize and quantify the collagen fiber (lamellar) organization of human corneas in three dimensions by using nonlinear optical high-resolution macroscopy (NLO-HRMac) and to correlate these findings with mechanical data obtained by indentation testing of corneal flaps. Methods. Twelve corneas from 10 donors were studied. Vibratome sections, 200 μm thick, from five donor eyes were cut along the vertical meridian from limbus to limbus (arc length, 12 mm). Backscattered second harmonic–generated (SHG) NLO signals from these sections were collected as a series of overlapping 3-D images, which were concatenated to form a single 3-D mosaic (pixel resolution: 0.44 μm lateral, 2 μm axial). Collagen fiber intertwining was quantified by determining branching point density as a function of stromal depth. Mechanical testing was performed on corneal flaps from seven additional eyes. Corneas were cut into three layers (anterior, middle, and posterior) using a femtosecond surgical laser system and underwent indentation testing to determine the elastic modulus for each layer. Results. The 3-D reconstructions revealed complex collagen fiber branching patterns in the anterior cornea, with fibers extending from the anterior limiting lamina (ALL, Bowman's layer), intertwining with deeper fibers and reinserting back to the ALL, forming bow spring–like structures. Measured branching-point density was four times higher in the anterior third of the cornea than in the posterior third and decreased logarithmically with increasing distance from the ALL. Indentation testing showed an eightfold increase in elastic modulus in the anterior stroma. Conclusions. The axial gradient in lamellar intertwining appears to be associated with an axial gradient in the effective elastic modulus of the cornea, suggesting that collagen fiber intertwining and formation of bow spring–like structures provide structural support similar to cross-beams in bridges and large-scale structures

  15. Monitoring synaptic and neuronal activity in 3D with synthetic and genetic indicators using a compact acousto-optic lens two-photon microscope☆

    PubMed Central

    Fernández-Alfonso, Tomás; Nadella, K.M. Naga Srinivas; Iacaruso, M. Florencia; Pichler, Bruno; Roš, Hana; Kirkby, Paul A.; Silver, R. Angus

    2014-01-01

    Background Two-photon microscopy is widely used to study brain function, but conventional microscopes are too slow to capture the timing of neuronal signalling and imaging is restricted to one plane. Recent development of acousto-optic-deflector-based random access functional imaging has improved the temporal resolution, but the utility of these technologies for mapping 3D synaptic activity patterns and their performance at the excitation wavelengths required to image genetically encoded indicators have not been investigated. New method Here, we have used a compact acousto-optic lens (AOL) two-photon microscope to make high speed [Ca2+] measurements from spines and dendrites distributed in 3D with different excitation wavelengths (800–920 nm). Results We show simultaneous monitoring of activity from many synaptic inputs distributed over the 3D arborisation of a neuronal dendrite using both synthetic as well as genetically encoded indicators. We confirm the utility of AOL-based imaging for fast in vivo recordings by measuring, simultaneously, visually evoked responses in 100 neurons distributed over a 150 μm focal depth range. Moreover, we explore ways to improve the measurement of timing of neuronal activation by choosing specific regions within the cell soma. Comparison with existing methods These results establish that AOL-based 3D random access two-photon microscopy has a wider range of neuroscience applications than previously shown. Conclusions Our findings show that the compact AOL microscope design has the speed, spatial resolution, sensitivity and wavelength flexibility to measure 3D patterns of synaptic and neuronal activity on individual trials. PMID:24200507

  16. Virtual k -Space Modulation Optical Microscopy

    NASA Astrophysics Data System (ADS)

    Kuang, Cuifang; Ma, Ye; Zhou, Renjie; Zheng, Guoan; Fang, Yue; Xu, Yingke; Liu, Xu; So, Peter T. C.

    2016-07-01

    We report a novel superresolution microscopy approach for imaging fluorescence samples. The reported approach, termed virtual k -space modulation optical microscopy (VIKMOM), is able to improve the lateral resolution by a factor of 2, reduce the background level, improve the optical sectioning effect and correct for unknown optical aberrations. In the acquisition process of VIKMOM, we used a scanning confocal microscope setup with a 2D detector array to capture sample information at each scanned x -y position. In the recovery process of VIKMOM, we first modulated the captured data by virtual k -space coding and then employed a ptychography-inspired procedure to recover the sample information and correct for unknown optical aberrations. We demonstrated the performance of the reported approach by imaging fluorescent beads, fixed bovine pulmonary artery endothelial (BPAE) cells, and living human astrocytes (HA). As the VIKMOM approach is fully compatible with conventional confocal microscope setups, it may provide a turn-key solution for imaging biological samples with ˜100 nm lateral resolution, in two or three dimensions, with improved optical sectioning capabilities and aberration correcting.

  17. High-Throughput Nonlinear Optical Microscopy

    PubMed Central

    So, Peter T.C.; Yew, Elijah Y.S.; Rowlands, Christopher

    2013-01-01

    High-resolution microscopy methods based on different nonlinear optical (NLO) contrast mechanisms are finding numerous applications in biology and medicine. While the basic implementations of these microscopy methods are relatively mature, an important direction of continuing technological innovation lies in improving the throughput of these systems. Throughput improvement is expected to be important for studying fast kinetic processes, for enabling clinical diagnosis and treatment, and for extending the field of image informatics. This review will provide an overview of the fundamental limitations on NLO microscopy throughput. We will further cover several important classes of high-throughput NLO microscope designs with discussions on their strengths and weaknesses and their key biomedical applications. Finally, this review will close with a perspective of potential future technological improvements in this field. PMID:24359736

  18. A near-field optical microscopy nanoarray

    SciTech Connect

    Semin, D.J.; Ambrose, W.P.; Goodwin, P.M.; Kwller, A.; Wendt, J.R.

    1996-12-31

    Multiplexing near-field scanning optical microscopy (NSOM) by the use of a nanoarray with parallel imaging is studied. The fabrication, characterization, and utilization of nanoarrays with {approximately} 100 nm diameter apertures spaced 500 nm center-to- center is presented. Extremely uniform nanoarrays with {approximately} 10{sup 8} apertures were fabricated by electron beam lithography and reactive ion etching. The nanoarrays were characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). In this paper we utilize these nanoarrays in a laser-illuminated microscope with parallel detection on a charge- coupled device (CCD). Detection of B-phycoerythrin (B-PE) molecules using near-field illumination is presented. In principle, our system can be used to obtain high lateral resolution NSOM images over a wide-field of view (e.g. 50-100 {mu}m) within seconds.

  19. A prototype fan-beam optical CT scanner for 3D dosimetry

    SciTech Connect

    Campbell, Warren G.; Rudko, D. A.; Braam, Nicolas A.; Jirasek, Andrew; Wells, Derek M.

    2013-06-15

    Purpose: The objective of this work is to introduce a prototype fan-beam optical computed tomography scanner for three-dimensional (3D) radiation dosimetry. Methods: Two techniques of fan-beam creation were evaluated: a helium-neon laser (HeNe, {lambda} = 543 nm) with line-generating lens, and a laser diode module (LDM, {lambda} = 635 nm) with line-creating head module. Two physical collimator designs were assessed: a single-slot collimator and a multihole collimator. Optimal collimator depth was determined by observing the signal of a single photodiode with varying collimator depths. A method of extending the dynamic range of the system is presented. Two sample types were used for evaluations: nondosimetric absorbent solutions and irradiated polymer gel dosimeters, each housed in 1 liter cylindrical plastic flasks. Imaging protocol investigations were performed to address ring artefacts and image noise. Two image artefact removal techniques were performed in sinogram space. Collimator efficacy was evaluated by imaging highly opaque samples of scatter-based and absorption-based solutions. A noise-based flask registration technique was developed. Two protocols for gel manufacture were examined. Results: The LDM proved advantageous over the HeNe laser due to its reduced noise. Also, the LDM uses a wavelength more suitable for the PRESAGE{sup TM} dosimeter. Collimator depth of 1.5 cm was found to be an optimal balance between scatter rejection, signal strength, and manufacture ease. The multihole collimator is capable of maintaining accurate scatter-rejection to high levels of opacity with scatter-based solutions (T < 0.015%). Imaging protocol investigations support the need for preirradiation and postirradiation scanning to reduce reflection-based ring artefacts and to accommodate flask imperfections and gel inhomogeneities. Artefact removal techniques in sinogram space eliminate streaking artefacts and reduce ring artefacts of up to {approx}40% in magnitude. The

  20. 3D visualization of TiO2 nanocrystals in mesoporous nanocomposite using energy filtered transmission electron microscopy tomography.

    PubMed

    Gondo, Takashi; Kasama, Takeshi; Kaneko, Kenji

    2014-11-01

    IntroductionMesoporous silica, SBA-15, is one of the best candidate for the supporting material of catalytic nanoparticles because of its relative large and controllable pore size and large specific surface area [1]. So far, various nanoparticles, such as Au, Pt and Pd, have been introduced into the pore for catalytic application [2]. The size of nanoparticles supported inside SBA-15 is restricted by that of the pore, and they are usually ranging from 2 nm and 50 nm in space.It is necessary to anchor the nanoparticles within pores to avoid segregation / sintering of them. However, it is difficult to anchor them within pores in the case of use of deposition-precipitation method due to extreme low iso-electric point (IEP) of silica (∼2). Therefore, TiO2 nanocrystals (IEP 6-8) were then introduced to anchor AuNPs [3].In this study, EFTEM tomography was applied to examine the effectiveness of TiO2 for AuNPs. Materials and methodAu/TiO2-SBA-15 was embedded into epoxy resin for electron microscopy and microtomed to about 30 nm thickness. EFTEM-tomography was operated at 120 kV and using Ti-L ionization edge via three-window method. Prior to EFTEM, STEM-HAADF tomography was also carried out for visualizing AuNPs and for comparison. Result and discussionFigure 1 shows 3D-volume of AuNPs and TiO2 nanocrystals from EFTEM-tomography. TiO2 nanocrystals in the porous material were successfully visualized using EFTEM -tomography, and local relationship between AuNPs and TiO2 nanocrystals were revealed. A large number of TiO2 nanocrystals were randomly distributed in the SBA-15. It was found that most AuNPs were directly on the exposed TiO2 nanocrystals. It implies that TiO2 nanocrystals were exposed on the surface of the pore and anchored AuNPs inside the pores.jmicro;63/suppl_1/i27/DFU081F1F1DFU081F1Fig. 1.3D volume of AuNPs and TiO2 nanocrystals.

  1. 2D and 3D micro-XRF based on polycapillary optics at XLab Frascati

    NASA Astrophysics Data System (ADS)

    Polese, C.; Cappuccio, G.; Dabagov, S. B.; Hampai, D.; Liedl, A.; Pace, E.

    2015-08-01

    XRF imaging spectrometry is a powerful tool for materials characterization. A high spatial resolution is often required, in order to appreciate very tiny details of the studied object. With respect to simple pinholes, polycapillary optics allows much more intense fluxes to be achieved. This is fundamental to detect elements in trace and to strongly reduce the global acquisition time that is actually among the main reasons, in addition to radioprotection issues, affecting the competitiveness of XRF imaging with respect to other faster imaging techniques such as multispectral imaging. Unlike other well-known X-ray optics, principally employed for high brilliant radiation source such as synchrotron facilities, polyCO can be efficiently coupled also with conventional X-ray tubes. All these aspects make them the most suitable choice to realize portable, safe and high performing μXRF spectrometers. In this work preliminary results achieved with a novel 2D and 3D XRF facility, called Rainbow X-Ray (RXR), are reported, with particular attention to the spatial resolution achieved. RXR is based on the confocal arrangement of three polycapillary lenses, one focusing the primary beam and the other two capturing the fluorescence signal. The detection system is split in two couples of lens-detector in order to cover a wider energy range. The entire device is a laboratory user-friendly facility and, though it allows measurements on medium-size objects, its dimensions do not preclude it to be transported for in situ analysis on request, thanks also to a properly shielded cabinet.

  2. Best fit refractive index of matching liquid for 3D NIPAM gel dosimeters using optical CT

    NASA Astrophysics Data System (ADS)

    Chen, Chin-Hsing; Wu, Jay; Hsieh, Bor-Tsung; Chen, De-Shiou; Wang, Tzu-Hwei; Chien, Sou-Hsin; Chang, Yuan-Jen

    2014-11-01

    The accuracy of an optical computed tomography (CT)-based dosimeter is significantly affected by the refractive index (RI) of the matching liquid. Mismatched RI induces reflection and refraction as the laser beam passes through the gel phantom. Moreover, the unwanted light rays collected by the photodetector produce image artifacts after image reconstruction from the collected data. To obtain the best image quality, this study investigates the best-fit RI of the matching liquid for a 3D NIPAM gel dosimeter. The three recipes of NIPAM polymer gel used in this study consisted of 5% gelatin, 5% NIPAM and 3% N,N'-methylene bisacrylamide, which were combined with three compositions (5, 10, and 20 mM) of Tetrakis (hydroxymethyl) phosphonium chloride. Results were evaluated using a quantitative evaluation method of the gamma evaluation technique. Results showed that the best-fit RI for the non-irradiated NIPAM gel ranges from 1.340 to 1.346 for various NIPAM recipes with sensitivities ranging from 0.0113 to 0.0227. The greatest pass rate of 88.00% is achieved using best-fit RI=1.346 of the matching liquid. The adoption of mismatching RI decreases the gamma pass rate by 2.63% to 16.75% for all three recipes of NIPAM gel dosimeters. In addition, the maximum average deviation is less than 0.1% for the red and transparent matching liquids. Thus, the color of the matching liquid does not affect the measurement accuracy of the NIPAM gel dosimeter, as measured by optical CT.

  3. A new technique of recognition for coded targets in optical 3D measurement

    NASA Astrophysics Data System (ADS)

    Guo, Changye; Cheng, Xiaosheng; Cui, Haihua; Dai, Ning; Weng, Jinping

    2014-11-01

    A new technique for coded targets recognition in optical 3D-measurement application is proposed in this paper. Traditionally, point cloud registration is based on homologous features, such as the curvature, which is time-consuming and not reliable. For this, we paste some coded targets onto the surface of the object to be measured to improve the optimum target location and accurate correspondence among multi-source images. Circular coded targets are used, and an algorithm to automatically detecting them is proposed. This algorithm extracts targets with intensive bimodal histogram features from complex background, and filters noise according to their size, shape and intensity. In addition, the coded targets' identification is conducted out by their ring codes. We affine them around the circle inversely, set foreground and background respectively as 1 and 0 to constitute a binary number, and finally shift one bit every time to calculate a decimal one of the binary number to determine a minimum decimal number as its code. In this 3Dmeasurement application, we build a mutual relationship between different viewpoints containing three or more coded targets with different codes. Experiments show that it is of efficiency to obtain global surface data of an object to be measured and is robust to the projection angles and noise.

  4. Combining 3D optical imaging and dual energy absorptiometry to measure three compositional components.

    PubMed

    Malkov, Serghei; Shepherd, John

    2014-02-17

    We report on the design of the technique combining 3D optical imaging and dual-energy absorptiometry body scanning to estimate local body area compositions of three compartments. Dual-energy attenuation and body shape measures are used together to solve for the three compositional tissue thicknesses: water, lipid, and protein. We designed phantoms with tissue-like properties as our reference standards for calibration purposes. The calibration was created by fitting phantom values using non-linear regression of quadratic and truncated polynomials. Dual-energy measurements were performed on tissue-mimicking phantoms using a bone densitometer unit. The phantoms were made of materials shown to have similar x-ray attenuation properties of the biological compositional compartments. The components for the solid phantom were tested and their high energy/low energy attenuation ratios are in good correspondent to water, lipid, and protein for the densitometer x-ray region. The three-dimensional body shape was reconstructed from the depth maps generated by Microsoft Kinect for Windows. We used open-source Point Cloud Library and freeware software to produce dense point clouds. Accuracy and precision of compositional and thickness measures were calculated. The error contributions due to two modalities were estimated. The preliminary phantom composition and shape measurements are found to demonstrate the feasibility of the method proposed.

  5. Automated multilayer segmentation and characterization in 3D spectral-domain optical coherence tomography images

    NASA Astrophysics Data System (ADS)

    Hu, Zhihong; Wu, Xiaodong; Hariri, Amirhossein; Sadda, SriniVas R.

    2013-03-01

    Spectral-domain optical coherence tomography (SD-OCT) is a 3-D imaging technique, allowing direct visualization of retinal morphology and architecture. The various layers of the retina may be affected differentially by various diseases. In this study, an automated graph-based multilayer approach was developed to sequentially segment eleven retinal surfaces including the inner retinal bands to the outer retinal bands in normal SD-OCT volume scans at three different stages. For stage 1, the four most detectable and/or distinct surfaces were identified in the four-times-downsampled images and were used as a priori positional information to limit the graph search for other surfaces at stage 2. Eleven surfaces were then detected in the two-times-downsampled images at stage 2, and refined in the original image space at stage 3 using the graph search integrating the estimated morphological shape models. Twenty macular SD-OCT (Heidelberg Spectralis) volume scans from 20 normal subjects (one eye per subject) were used in this study. The overall mean and absolute mean differences in border positions between the automated and manual segmentation for all 11 segmented surfaces were -0.20 +/- 0.53 voxels (-0.76 +/- 2.06 μm) and 0.82 +/- 0.64 voxels (3.19 +/- 2.46 μm). Intensity and thickness properties in the resultant retinal layers were investigated. This investigation in normal subjects may provide a comparative reference for subsequent investigations in eyes with disease.

  6. Combining 3D optical imaging and dual energy absorptiometry to measure three compositional components

    PubMed Central

    Malkov, Serghei; Shepherd, John

    2014-01-01

    We report on the design of the technique combining 3D optical imaging and dual-energy absorptiometry body scanning to estimate local body area compositions of three compartments. Dual-energy attenuation and body shape measures are used together to solve for the three compositional tissue thicknesses: water, lipid, and protein. We designed phantoms with tissue-like properties as our reference standards for calibration purposes. The calibration was created by fitting phantom values using non-linear regression of quadratic and truncated polynomials. Dual-energy measurements were performed on tissue-mimicking phantoms using a bone densitometer unit. The phantoms were made of materials shown to have similar x-ray attenuation properties of the biological compositional compartments. The components for the solid phantom were tested and their high energy/low energy attenuation ratios are in good correspondent to water, lipid, and protein for the densitometer x-ray region. The three-dimensional body shape was reconstructed from the depth maps generated by Microsoft Kinect for Windows. We used open-source Point Cloud Library and freeware software to produce dense point clouds. Accuracy and precision of compositional and thickness measures were calculated. The error contributions due to two modalities were estimated. The preliminary phantom composition and shape measurements are found to demonstrate the feasibility of the method proposed. PMID:25083118

  7. Combining 3D optical imaging and dual energy absorptiometry to measure three compositional components

    NASA Astrophysics Data System (ADS)

    Malkov, Serghei; Shepherd, John

    2014-02-01

    We report on the design of the technique combining 3D optical imaging and dual-energy absorptiometry body scanning to estimate local body area compositions of three compartments. Dual-energy attenuation and body shape measures are used together to solve for the three compositional tissue thicknesses: water, lipid, and protein. We designed phantoms with tissue-like properties as our reference standards for calibration purposes. The calibration was created by fitting phantom values using non-linear regression of quadratic and truncated polynomials. Dual-energy measurements were performed on tissue-mimicking phantoms using a bone densitometer unit. The phantoms were made of materials shown to have similar x-ray attenuation properties of the biological compositional compartments. The components for the solid phantom were tested and their high energy/low energy attenuation ratios are in good correspondent to water, lipid, and protein for the densitometer x-ray region. The three-dimensional body shape was reconstructed from the depth maps generated by Microsoft Kinect for Windows. We used open-source Point Cloud Library and freeware software to produce dense point clouds. Accuracy and precision of compositional and thickness measures were calculated. The error contributions due to two modalities were estimated. The preliminary phantom composition and shape measurements are found to demonstrate the feasibility of the method proposed.

  8. Large area 3-D optical coherence tomography imaging of lumpectomy specimens for radiation treatment planning

    NASA Astrophysics Data System (ADS)

    Wang, Cuihuan; Kim, Leonard; Barnard, Nicola; Khan, Atif; Pierce, Mark C.

    2016-02-01

    Our long term goal is to develop a high-resolution imaging method for comprehensive assessment of tissue removed during lumpectomy procedures. By identifying regions of high-grade disease within the excised specimen, we aim to develop patient-specific post-operative radiation treatment regimens. We have assembled a benchtop spectral-domain optical coherence tomography (SD-OCT) system with 1320 nm center wavelength. Automated beam scanning enables "sub-volumes" spanning 5 mm x 5 mm x 2 mm (500 A-lines x 500 B-scans x 2 mm in depth) to be collected in under 15 seconds. A motorized sample positioning stage enables multiple sub-volumes to be acquired across an entire tissue specimen. Sub-volumes are rendered from individual B-scans in 3D Slicer software and en face (XY) images are extracted at specific depths. These images are then tiled together using MosaicJ software to produce a large area en face view (up to 40 mm x 25 mm). After OCT imaging, specimens were sectioned and stained with HE, allowing comparison between OCT image features and disease markers on histopathology. This manuscript describes the technical aspects of image acquisition and reconstruction, and reports initial qualitative comparison between large area en face OCT images and HE stained tissue sections. Future goals include developing image reconstruction algorithms for mapping an entire sample, and registering OCT image volumes with clinical CT and MRI images for post-operative treatment planning.

  9. Optical absorption enhancement in 3D silicon oxide nano-sandwich type solar cell.

    PubMed

    Kiani, Amirkianoosh; Venkatakrishnan, Krishnan; Tan, Bo

    2014-01-13

    Recent research in the field of photovoltaic and solar cell fabrication has shown the potential to significantly enhance light absorption in thin-film solar cells by using surface texturing and nanostructure coating techniques. In this paper, for the first time, we propose a new method for nano sandwich type thin-film solar cell fabrication by combining the laser amorphization (2nd solar cell generation) and laser nanofibers generation (3rd solar cell generation) techniques. In this novel technique, the crystalline silicon is irradiated by megahertz frequency femtosecond laser pulses under ambient conditions and the multi-layer of amorphorized silicon and nano fibrous layer are generated in the single-step on top of the silicon substrate. Light spectroscopy results show significant enhancement of light absorption in the generated multi layers solar cells (Silicon Oxide nanofibers / thin-film amorphorized silicon). This method is single step and no additional materials are added and both layers of the amorphorized thin-film silicon and three-dimensional (3D) silicon oxide nanofibrous structures are grown on top of the silicon substrate after laser irradiation. Finally, we suggest how to maximize the light trapping and optical absorption of the generated nanofibers/thin-film cells by optimizing the laser pulse duration. PMID:24921988

  10. Detection of latent fingerprints using high-resolution 3D confocal microscopy in non-planar acquisition scenarios

    NASA Astrophysics Data System (ADS)

    Kirst, Stefan; Vielhauer, Claus

    2015-03-01

    In digitized forensics the support of investigators in any manner is one of the main goals. Using conservative lifting methods, the detection of traces is done manually. For non-destructive contactless methods, the necessity for detecting traces is obvious for further biometric analysis. High resolutional 3D confocal laser scanning microscopy (CLSM) grants the possibility for a detection by segmentation approach with improved detection results. Optimal scan results with CLSM are achieved on surfaces orthogonal to the sensor, which is not always possible due to environmental circumstances or the surface's shape. This introduces additional noise, outliers and a lack of contrast, making a detection of traces even harder. Prior work showed the possibility of determining angle-independent classification models for the detection of latent fingerprints (LFP). Enhancing this approach, we introduce a larger feature space containing a variety of statistical-, roughness-, color-, edge-directivity-, histogram-, Gabor-, gradient- and Tamura features based on raw data and gray-level co-occurrence matrices (GLCM) using high resolutional data. Our test set consists of eight different surfaces for the detection of LFP in four different acquisition angles with a total of 1920 single scans. For each surface and angles in steps of 10, we capture samples from five donors to introduce variance by a variety of sweat compositions and application influences such as pressure or differences in ridge thickness. By analyzing the present test set with our approach, we intend to determine angle- and substrate-dependent classification models to determine optimal surface specific acquisition setups and also classification models for a general detection purpose for both, angles and substrates. The results on overall models with classification rates up to 75.15% (kappa 0.50) already show a positive tendency regarding the usability of the proposed methods for LFP detection on varying surfaces in non

  11. Porosity and permeability determination of organic-rich Posidonia shales based on 3-D analyses by FIB-SEM microscopy

    NASA Astrophysics Data System (ADS)

    Grathoff, Georg H.; Peltz, Markus; Enzmann, Frieder; Kaufhold, Stephan

    2016-07-01

    The goal of this study is to better understand the porosity and permeability in shales to improve modelling fluid and gas flow related to shale diagenesis. Two samples (WIC and HAD) were investigated, both mid-Jurassic organic-rich Posidonia shales from Hils area, central Germany of different maturity (WIC R0 0.53 % and HAD R0 1.45 %). The method for image collection was focused ion beam (FIB) microscopy coupled with scanning electron microscopy (SEM). For image and data analysis Avizo and GeoDict was used. Porosity was calculated from segmented 3-D FIB based images and permeability was simulated by a Navier Stokes-Brinkman solver in the segmented images. Results show that the quantity and distribution of pore clusters and pores (≥ 40 nm) are similar. The largest pores are located within carbonates and clay minerals, whereas the smallest pores are within the matured organic matter. Orientation of the pores calculated as pore paths showed minor directional differences between the samples. Both samples have no continuous connectivity of pore clusters along the axes in the x, y, and z direction on the scale of 10 to 20 of micrometer, but do show connectivity on the micrometer scale. The volume of organic matter in the studied volume is representative of the total organic carbon (TOC) in the samples. Organic matter does show axis connectivity in the x, y, and z directions. With increasing maturity the porosity in organic matter increases from close to 0 to more than 5 %. These pores are small and in the large organic particles have little connection to the mineral matrix. Continuous pore size distributions are compared with mercury intrusion porosimetry (MIP) data. Differences between both methods are caused by resolution limits of the FIB-SEM and by the development of small pores during the maturation of the organic matter. Calculations show no permeability when only considering visible pores due to the lack of axis connectivity. Adding the organic matter with a

  12. Visualizing the 3D Architecture of Multiple Erythrocytes Infected with Plasmodium at Nanoscale by Focused Ion Beam-Scanning Electron Microscopy

    PubMed Central

    Soares Medeiros, Lia Carolina; De Souza, Wanderley; Jiao, Chengge; Barrabin, Hector; Miranda, Kildare

    2012-01-01

    Different methods for three-dimensional visualization of biological structures have been developed and extensively applied by different research groups. In the field of electron microscopy, a new technique that has emerged is the use of a focused ion beam and scanning electron microscopy for 3D reconstruction at nanoscale resolution. The higher extent of volume that can be reconstructed with this instrument represent one of the main benefits of this technique, which can provide statistically relevant 3D morphometrical data. As the life cycle of Plasmodium species is a process that involves several structurally complex developmental stages that are responsible for a series of modifications in the erythrocyte surface and cytoplasm, a high number of features within the parasites and the host cells has to be sampled for the correct interpretation of their 3D organization. Here, we used FIB-SEM to visualize the 3D architecture of multiple erythrocytes infected with Plasmodium chabaudi and analyzed their morphometrical parameters in a 3D space. We analyzed and quantified alterations on the host cells, such as the variety of shapes and sizes of their membrane profiles and parasite internal structures such as a polymorphic organization of hemoglobin-filled tubules. The results show the complex 3D organization of Plasmodium and infected erythrocyte, and demonstrate the contribution of FIB-SEM for the obtainment of statistical data for an accurate interpretation of complex biological structures. PMID:22432024

  13. Dark-field optical coherence microscopy

    NASA Astrophysics Data System (ADS)

    Pache, C.; Villiger, M. L.; Lasser, T.

    2010-02-01

    Many solutions have been proposed to produce phase quantitative images of biological cell samples. Among these, Spectral Domain Phase Microscopy combines the fast imaging speed and high sensitivity of Optical Coherence Microscopy (OCM) in the Fourier domain with the high phase stability of common-path interferometry. We report on a new illumination scheme for OCM that enhances the sensitivity for backscattered light and detects the weak sample signal, otherwise buried by the signal from specular reflection. With the use of a Bessel-like beam, a dark-field configuration was realized. Sensitivity measurements for three different illumination configurations were performed to compare our method to standard OCM and extended focus OCM. Using a well-defined scattering and reflecting object, we demonstrated an attenuation of -40 dB of the DC-component and a relative gain of 30 dB for scattered light, compared to standard OCM. In a second step, we applied this technique, referred to as dark-field Optical Coherence Microscopy (dfOCM), to living cells. Chinese hamster ovarian cells were applied in a drop of medium on a coverslide. The cells of ~15 μm in diameter and even internal cell structures were visualized in the acquired tomograms.

  14. Single-spin stochastic optical reconstruction microscopy

    PubMed Central

    Pfender, Matthias; Aslam, Nabeel; Waldherr, Gerald; Neumann, Philipp; Wrachtrup, Jörg

    2014-01-01

    We experimentally demonstrate precision addressing of single-quantum emitters by combined optical microscopy and spin resonance techniques. To this end, we use nitrogen vacancy (NV) color centers in diamond confined within a few ten nanometers as individually resolvable quantum systems. By developing a stochastic optical reconstruction microscopy (STORM) technique for NV centers, we are able to simultaneously perform sub–diffraction-limit imaging and optically detected spin resonance (ODMR) measurements on NV spins. This allows the assignment of spin resonance spectra to individual NV center locations with nanometer-scale resolution and thus further improves spatial discrimination. For example, we resolved formerly indistinguishable emitters by their spectra. Furthermore, ODMR spectra contain metrology information allowing for sub–diffraction-limit sensing of, for instance, magnetic or electric fields with inherently parallel data acquisition. As an example, we have detected nuclear spins with nanometer-scale precision. Finally, we give prospects of how this technique can evolve into a fully parallel quantum sensor for nanometer resolution imaging of delocalized quantum correlations. PMID:25267655

  15. Multiparallel Three-Dimensional Optical Microscopy

    NASA Technical Reports Server (NTRS)

    Nguyen, Lam K.; Price, Jeffrey H.; Kellner, Albert L.; Bravo-Zanoquera, Miguel

    2010-01-01

    Multiparallel three-dimensional optical microscopy is a method of forming an approximate three-dimensional image of a microscope sample as a collection of images from different depths through the sample. The imaging apparatus includes a single microscope plus an assembly of beam splitters and mirrors that divide the output of the microscope into multiple channels. An imaging array of photodetectors in each channel is located at a different distance along the optical path from the microscope, corresponding to a focal plane at a different depth within the sample. The optical path leading to each photodetector array also includes lenses to compensate for the variation of magnification with distance so that the images ultimately formed on all the photodetector arrays are of the same magnification. The use of optical components common to multiple channels in a simple geometry makes it possible to obtain high light-transmission efficiency with an optically and mechanically simple assembly. In addition, because images can be read out simultaneously from all the photodetector arrays, the apparatus can support three-dimensional imaging at a high scanning rate.

  16. Three-dimensional imaging of normal skin and nonmelanoma skin cancer with cellular resolution using Gabor domain optical coherence microscopy.

    PubMed

    Lee, Kye-Sung; Zhao, Huimin; Ibrahim, Sherrif F; Meemon, Natthani; Khoudeir, Laura; Rolland, Jannick P

    2012-12-01

    We investigate morphological differences in three-dimensional (3-D) images with cellular resolution between nonmelanoma skin cancer and normal skin using Gabor domain optical coherence microscopy. As a result, we show for the first time cellular optical coherence images of 3-D features differentiating cancerous skin from normal skin. In addition, in vivo volumetric images of normal skin from different anatomic locations are shown and compared.

  17. Creation of quantum-degenerate gases of ytterbium in a compact 2D-/3D-magneto-optical trap setup

    SciTech Connect

    Doerscher, Soeren; Thobe, Alexander; Hundt, Bastian; Kochanke, Andre; Le Targat, Rodolphe; Windpassinger, Patrick; Becker, Christoph; Sengstock, Klaus

    2013-04-15

    We report on the first experimental setup based on a 2D-/3D-magneto-optical trap (MOT) scheme to create both Bose-Einstein condensates and degenerate Fermi gases of several ytterbium isotopes. Our setup does not require a Zeeman slower and offers the flexibility to simultaneously produce ultracold samples of other atomic species. Furthermore, the extraordinary optical access favors future experiments in optical lattices. A 2D-MOT on the strong {sup 1}S{sub 0}{yields}{sup 1}P{sub 1} transition captures ytterbium directly from a dispenser of atoms and loads a 3D-MOT on the narrow {sup 1}S{sub 0}{yields}{sup 3}P{sub 1} intercombination transition. Subsequently, atoms are transferred to a crossed optical dipole trap and cooled evaporatively to quantum degeneracy.

  18. Implementation of PSF engineering in high-resolution 3D microscopy imaging with a LCoS (reflective) SLM

    NASA Astrophysics Data System (ADS)

    King, Sharon V.; Doblas, Ana; Patwary, Nurmohammed; Saavedra, Genaro; Martínez-Corral, Manuel; Preza, Chrysanthe

    2014-03-01

    Wavefront coding techniques are currently used to engineer unique point spread functions (PSFs) that enhance existing microscope modalities or create new ones. Previous work in this field demonstrated that simulated intensity PSFs encoded with a generalized cubic phase mask (GCPM) are invariant to spherical aberration or misfocus; dependent on parameter selection. Additional work demonstrated that simulated PSFs encoded with a squared cubic phase mask (SQUBIC) produce a depth invariant focal spot for application in confocal scanning microscopy. Implementation of PSF engineering theory with a liquid crystal on silicon (LCoS) spatial light modulator (SLM) enables validation of WFC phase mask designs and parameters by manipulating optical wavefront properties with a programmable diffractive element. To validate and investigate parameters of the GCPM and SQUBIC WFC masks, we implemented PSF engineering in an upright microscope modified with a dual camera port and a LCoS SLM. We present measured WFC PSFs and compare them to simulated PSFs through analysis of their effect on the microscope imaging system properties. Experimentally acquired PSFs show the same intensity distribution as simulation for the GCPM phase mask, the SQUBIC-mask and the well-known and characterized cubic-phase mask (CPM), first applied to high NA microscopy by Arnison et al.10, for extending depth of field. These measurements provide experimental validation of new WFC masks and demonstrate the use of the LCoS SLM as a WFC design tool. Although efficiency improvements are needed, this application of LCoS technology renders the microscope capable of switching among multiple WFC modes.

  19. Development of a strain analyzer system for sheet metal stamped parts based on an optical 3D digitizer

    NASA Astrophysics Data System (ADS)

    Carasusan, Eusebio; Canal, Fernando

    2003-05-01

    In this paper a new automatic strain analyzer system for sheet metal stamped parts is presented. This device is based on an optic 3D scanner working with structure white light, that performs measurements with high speed and accuracy. This new device can be a very useful tool in manufacturing industry.

  20. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy.

    PubMed

    Tyson, Adam L; Hilton, Stephen T; Andreae, Laura C

    2015-10-30

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods.

  1. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy

    PubMed Central

    Tyson, Adam L.; Hilton, Stephen T.; Andreae, Laura C.

    2015-01-01

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods. PMID:25797056

  2. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy.

    PubMed

    Tyson, Adam L; Hilton, Stephen T; Andreae, Laura C

    2015-10-30

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods. PMID:25797056

  3. Characterization of 3D printing output using an optical sensing system

    NASA Astrophysics Data System (ADS)

    Straub, Jeremy

    2015-05-01

    This paper presents the experimental design and initial testing of a system to characterize the progress and performance of a 3D printer. The system is based on five Raspberry Pi single-board computers. It collects images of the 3D printed object, which are compared to an ideal model. The system, while suitable for printers of all sizes, can potentially be produced at a sufficiently low cost to allow its incorporation into consumer-grade printers. The efficacy and accuracy of this system is presented and discussed. The paper concludes with a discussion of the benefits of being able to characterize 3D printer performance.

  4. Analysis of 3D-printed metal for rapid-prototyped reflective terahertz optics.

    PubMed

    Headland, Daniel; Withayachumnankul, Withawat; Webb, Michael; Ebendorff-Heidepriem, Heike; Luiten, Andre; Abbott, Derek

    2016-07-25

    We explore the potential of 3D metal printing to realize complex conductive terahertz devices. Factors impacting performance such as printing resolution, surface roughness, oxidation, and material loss are investigated via analytical, numerical, and experimental approaches. The high degree of control offered by a 3D-printed topology is exploited to realize a zone plate operating at 530 GHz. Reflection efficiency at this frequency is found to be over 90%. The high-performance of this preliminary device suggest that 3D metal printing can play a strong role in guided-wave and general beam control devices in the terahertz range. PMID:27464185

  5. Analysis of 3D-printed metal for rapid-prototyped reflective terahertz optics

    NASA Astrophysics Data System (ADS)

    Headland, Daniel; Withayachumnankul, Withawat; Webb, Michael; Ebendorff-Heidepriem, Heike; Luiten, Andre; Abbott, Derek

    2016-07-01

    We explore the potential of 3D metal printing to realize complex conductive terahertz devices. Factors impacting performance such as printing resolution, surface roughness, oxidation, and material loss are investigated via analytical, numerical, and experimental approaches. The high degree of control offered by a 3D-printed topology is exploited to realize a zone plate operating at 530 GHz. Reflection efficiency at this frequency is found to be over 90%. The high-performance of this preliminary device suggest that 3D metal printing can play a strong role in guided-wave and general beam control devices in the terahertz range.

  6. Analysis of 3D-printed metal for rapid-prototyped reflective terahertz optics.

    PubMed

    Headland, Daniel; Withayachumnankul, Withawat; Webb, Michael; Ebendorff-Heidepriem, Heike; Luiten, Andre; Abbott, Derek

    2016-07-25

    We explore the potential of 3D metal printing to realize complex conductive terahertz devices. Factors impacting performance such as printing resolution, surface roughness, oxidation, and material loss are investigated via analytical, numerical, and experimental approaches. The high degree of control offered by a 3D-printed topology is exploited to realize a zone plate operating at 530 GHz. Reflection efficiency at this frequency is found to be over 90%. The high-performance of this preliminary device suggest that 3D metal printing can play a strong role in guided-wave and general beam control devices in the terahertz range.

  7. 3D Imaging of Porous Media Using Laser Scanning Confocal Microscopy with Application to Microscale Transport Processes

    SciTech Connect

    Fredrich, J.T.

    1999-02-10

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

  8. High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens

    NASA Astrophysics Data System (ADS)

    Philipp, Katrin; Smolarski, André; Fischer, Andreas; Koukourakis, Nektarios; Stürmer, Moritz; Wallrabe, Ulricke; Czarske, Jürgen

    2016-04-01

    We present a HiLo microscope with an electrically tunable lens for high-contrast three-dimensional image acquisition. HiLo microscopy combines wide field and speckled illumination images to create optically sectioned images. Additionally, the depth-of-field is not fixed, but can be adjusted between wide field and confocal-like axial resolution. We incorporate an electrically tunable lens in the HiLo microscope for axial scanning, to obtain three-dimensional data without the need of moving neither the sample nor the objective. The used adaptive lens consists of a transparent polydimethylsiloxane (PDMS) membrane into which an annular piezo bending actuator is embedded. A transparent fluid is filled between the membrane and the glass substrate. When actuated, the piezo generates a pressure in the lens which deflects the membrane and thus changes the refractive power. This technique enables a large tuning range of the refractive power between 1/f = (-24 . . . 25) 1/m. As the NA of the adaptive lens is only about 0.05, a fixed high-NA lens is included in the setup to provide high resolution. In this contribution, the scan properties and capabilities of the tunable lens in the HiLo microscope are analyzed. Eventually, exemplary measurements are presented and discussed.

  9. 3D imaging of cone photoreceptors over extended time periods using optical coherence tomography with adaptive optics

    NASA Astrophysics Data System (ADS)

    Kocaoglu, Omer P.; Lee, Sangyeol; Jonnal, Ravi S.; Wang, Qiang; Herde, Ashley E.; Besecker, Jason; Gao, Weihua; Miller, Donald T.

    2011-03-01

    Optical coherence tomography with adaptive optics (AO-OCT) is a highly sensitive, noninvasive method for 3D imaging of the microscopic retina. The purpose of this study is to advance AO-OCT technology by enabling repeated imaging of cone photoreceptors over extended periods of time (days). This sort of longitudinal imaging permits monitoring of 3D cone dynamics in both normal and diseased eyes, in particular the physiological processes of disc renewal and phagocytosis, which are disrupted by retinal diseases such as age related macular degeneration and retinitis pigmentosa. For this study, the existing AO-OCT system at Indiana underwent several major hardware and software improvements to optimize system performance for 4D cone imaging. First, ultrahigh speed imaging was realized using a Basler Sprint camera. Second, a light source with adjustable spectrum was realized by integration of an Integral laser (Femto Lasers, λc=800nm, ▵λ=160nm) and spectral filters in the source arm. For cone imaging, we used a bandpass filter with λc=809nm and ▵λ=81nm (2.6 μm nominal axial resolution in tissue, and 167 KHz A-line rate using 1,408 px), which reduced the impact of eye motion compared to previous AO-OCT implementations. Third, eye motion artifacts were further reduced by custom ImageJ plugins that registered (axially and laterally) the volume videos. In two subjects, cone photoreceptors were imaged and tracked over a ten day period and their reflectance and outer segment (OS) lengths measured. High-speed imaging and image registration/dewarping were found to reduce eye motion to a fraction of a cone width (1 μm root mean square). The pattern of reflections in the cones was found to change dramatically and occurred on a spatial scale well below the resolution of clinical instruments. Normalized reflectance of connecting cilia (CC) and OS posterior tip (PT) of an exemplary cone was 54+/-4, 47+/-4, 48+/-6, 50+/-5, 56+/-1% and 46+/-4, 53+/-4, 52+/-6, 50+/-5, 44

  10. 3-D segmentation of retinal blood vessels in spectral-domain OCT volumes of the optic nerve head

    NASA Astrophysics Data System (ADS)

    Lee, Kyungmoo; Abràmoff, Michael D.; Niemeijer, Meindert; Garvin, Mona K.; Sonka, Milan

    2010-03-01

    Segmentation of retinal blood vessels can provide important information for detecting and tracking retinal vascular diseases including diabetic retinopathy, arterial hypertension, arteriosclerosis and retinopathy of prematurity (ROP). Many studies on 2-D segmentation of retinal blood vessels from a variety of medical images have been performed. However, 3-D segmentation of retinal blood vessels from spectral-domain optical coherence tomography (OCT) volumes, which is capable of providing geometrically accurate vessel models, to the best of our knowledge, has not been previously studied. The purpose of this study is to develop and evaluate a method that can automatically detect 3-D retinal blood vessels from spectral-domain OCT scans centered on the optic nerve head (ONH). The proposed method utilized a fast multiscale 3-D graph search to segment retinal surfaces as well as a triangular mesh-based 3-D graph search to detect retinal blood vessels. An experiment on 30 ONH-centered OCT scans (15 right eye scans and 15 left eye scans) from 15 subjects was performed, and the mean unsigned error in 3-D of the computer segmentations compared with the independent standard obtained from a retinal specialist was 3.4 +/- 2.5 voxels (0.10 +/- 0.07 mm).

  11. Optical low-cost and portable arrangement for full field 3D displacement measurement using a single camera

    NASA Astrophysics Data System (ADS)

    López-Alba, E.; Felipe-Sesé, L.; Schmeer, S.; Díaz, F. A.

    2016-11-01

    In the current paper, an optical low-cost system for 3D displacement measurement based on a single camera and 3D digital image correlation is presented. The conventional 3D-DIC set-up based on a two-synchronized-cameras system is compared with a proposed pseudo-stereo portable system that employs a mirror system integrated in a device for a straightforward application achieving a novel handle and flexible device for its use in many scenarios. The proposed optical system splits the image by the camera into two stereo images of the object. In order to validate this new approach and quantify its uncertainty compared to traditional 3D-DIC systems, solid rigid in and out-of-plane displacements experiments have been performed and analyzed. The differences between both systems have been studied employing an image decomposition technique which performs a full image comparison. Therefore, results of all field of view are compared with those using a stereoscopy system and 3D-DIC, discussing the accurate results obtained with the proposed device not having influence any distortion or aberration produced by the mirrors. Finally, the adaptability of the proposed system and its accuracy has been tested performing quasi-static and dynamic experiments using a silicon specimen under high deformation. Results have been compared and validated with those obtained from a conventional stereoscopy system showing an excellent level of agreement.

  12. Structured light optical microscopy for three-dimensional reconstruction of technical surfaces

    NASA Astrophysics Data System (ADS)

    Kettel, Johannes; Reinecke, Holger; Müller, Claas

    2016-04-01

    In microsystems technology quality control of micro structured surfaces with different surface properties is playing an ever more important role. The process of quality control incorporates three-dimensional (3D) reconstruction of specularand diffusive reflecting technical surfaces. Due to the demand on high measurement accuracy and data acquisition rates, structured light optical microscopy has become a valuable solution to solve this problem providing high vertical and lateral resolution. However, 3D reconstruction of specular reflecting technical surfaces still remains a challenge to optical measurement principles. In this paper we present a measurement principle based on structured light optical microscopy which enables 3D reconstruction of specular- and diffusive reflecting technical surfaces. It is realized using two light paths of a stereo microscope equipped with different magnification levels. The right optical path of the stereo microscope is used to project structured light onto the object surface. The left optical path is used to capture the structured illuminated object surface with a camera. Structured light patterns are generated by a Digital Light Processing (DLP) device in combination with a high power Light Emitting Diode (LED). Structured light patterns are realized as a matrix of discrete light spots to illuminate defined areas on the object surface. The introduced measurement principle is based on multiple and parallel processed point measurements. Analysis of the measured Point Spread Function (PSF) by pattern recognition and model fitting algorithms enables the precise calculation of 3D coordinates. Using exemplary technical surfaces we demonstrate the successful application of our measurement principle.

  13. DMD-based LED-illumination super-resolution and optical sectioning microscopy.

    PubMed

    Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

    2013-01-01

    Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×10(7) pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

  14. DMD-based LED-illumination Super-resolution and optical sectioning microscopy

    NASA Astrophysics Data System (ADS)

    Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

    2013-01-01

    Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×107 pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

  15. Nondestructive testing of 3D disperse systems with micro- and nano-particles: N-dimensional space of optical parameters

    NASA Astrophysics Data System (ADS)

    Bezrukova, Alexandra G.

    2006-04-01

    The simultaneous analysis of 3D disperse systems (DS) with micro- and nano- particles by refractometry, absorbency, fluorescence and by different types of light scattering, can help to elaborate the sensing elements for specffic impurity control. Our research has investigated by complex of optical methods different 3D DS such as: proteins, nucleoproteids, lipoproteids, liposomes, viruses, virosomes, lipid emulsions, blood substitutes, latexes, liquid crystals, biological cells with various form and size (including bacterial cells), metallic powders, clays, kimberlites, zeolites, oils, crude oils, samples of natural and water-supply waters, etc. This experience suggests that each 3D DS can be charactensed by N-dimensional vector in N-dimensional space of optical parameters. Due to the fusion of various optical data it is possible to solve the inverse physical problem on the presence of impurity in mixtures of 3D DS by information statistical theory methods. It is important that in this case polymodality of particle size distribution is not an obstacle.

  16. Combining confocal microscopy with precise force-scope optical tweezers

    NASA Astrophysics Data System (ADS)

    Richardson, Andrew C.; Reihani, Nader; Oddershede, Lene B.

    2006-08-01

    We demonstrate an example of 'confocal-tweezers' wherein confocal images and precise optical force measurements, using photodiodes, are obtained simultaneously in the x-y plane without moving the objective lens. The optical trap is produced using a 1.064μm cw laser and is combined with Leica's TCS SP5 broadband confocal microscope to trap and image living cells. The unique method by which the confocal images are created facilitates the acquisition of images in areas far from the trapping location. In addition, because the scanning process involves moving galvanic mirrors independently of the objective, the trap is held stable in position and is not subject to any error in position for the x-y scan. We have successfully trapped and confocally imaged 80nm gold colloids, 150nm gold colloids and 1μm polystyrene beads whilst making quantitative measurements of the force applied by the trap on each bead. To the best of our knowledge this is the first time that anyone has combined precise force measuring optical tweezers with confocal microscopy. We also discuss some of the technical challenges involved in advancing the experimental set up to make quantitative force measurements in combination with 3D stacking. Having proven the potential of this system in 2D, we hope to develop it further to investigate the nano-mechanics of cell division through the attachment of gold beads to fluorescently labelled organelles in S. pombe yeast cells.

  17. Measurements of adipose derived stem cell vitality with optical coherence phase microscopy

    NASA Astrophysics Data System (ADS)

    Bagnaninchi, P. O.; Holmes, C.; Drummond, N.; Daoud, J.; Tabrizian, M.

    2011-03-01

    Live cells display a constant vertical motility due partly to a constant rearrangement of focal contacts and to cell shape fluctuations. This cellular micromotion has been clearly demonstrated with electric cell impedance sensing (ECIS) on 2D micro-electrodes, and correlated to cell vitality. In this study we investigated if optical coherence phase microscopy (OCPM) was able to report phase fluctuations of adult stem cells in 2D and 3D that could be correlated to cell motility. An OCPM has been developed around a Thorlabs engine (λο=930nm FWHM: 90nm) and integrated in an inverted microscope with a custom scanning head. Human adipose derived stem cells (ADSCs, Invitrogen) were cultured in Mesenpro RS medium and seeded either on ECIS arrays, 2D cell culture dishes, or in 3D highly porous microplotted polymeric scaffolds. ADSC motility was measured by ECIS and a spectral analysis was performed to retrieve the power spectral density (PSD) of the fluctuations. Cells in standard media and fixed cells were investigated. The same conditions were then investigated for ADSCs in 2D and in 3D with optical coherence phase microscopy. Significant differences were found in phase fluctuations between the different conditions, which correlated well with ECIS experiments. These preliminary results indicated that optical coherence phase microscopy could assess cell vitality in 2D and potentially in 3D microstructures.

  18. Remote z-scanning with a macroscopic voice coil motor for fast 3D multiphoton laser scanning microscopy

    PubMed Central

    Rupprecht, Peter; Prendergast, Andrew; Wyart, Claire; Friedrich, Rainer W

    2016-01-01

    There is a high demand for 3D multiphoton imaging in neuroscience and other fields but scanning in axial direction presents technical challenges. We developed a focusing technique based on a remote movable mirror that is conjugate to the specimen plane and translated by a voice coil motor. We constructed cost-effective z-scanning modules from off-the-shelf components that can be mounted onto standard multiphoton laser scanning microscopes to extend scan patterns from 2D to 3D. Systems were designed for large objectives and provide high resolution, high speed and a large z-scan range (>300 μm). We used these systems for 3D multiphoton calcium imaging in the adult zebrafish brain and measured odor-evoked activity patterns across >1500 neurons with single-neuron resolution and high signal-to-noise ratio. PMID:27231612

  19. Remote z-scanning with a macroscopic voice coil motor for fast 3D multiphoton laser scanning microscopy.

    PubMed

    Rupprecht, Peter; Prendergast, Andrew; Wyart, Claire; Friedrich, Rainer W

    2016-05-01

    There is a high demand for 3D multiphoton imaging in neuroscience and other fields but scanning in axial direction presents technical challenges. We developed a focusing technique based on a remote movable mirror that is conjugate to the specimen plane and translated by a voice coil motor. We constructed cost-effective z-scanning modules from off-the-shelf components that can be mounted onto standard multiphoton laser scanning microscopes to extend scan patterns from 2D to 3D. Systems were designed for large objectives and provide high resolution, high speed and a large z-scan range (>300 μm). We used these systems for 3D multiphoton calcium imaging in the adult zebrafish brain and measured odor-evoked activity patterns across >1500 neurons with single-neuron resolution and high signal-to-noise ratio. PMID:27231612

  20. Integrated optical coherence tomography and optical coherence microscopy imaging of human pathology

    NASA Astrophysics Data System (ADS)

    Lee, Hsiang-Chieh; Zhou, Chao; Wang, Yihong; Aquirre, Aaron D.; Tsai, Tsung-Han; Cohen, David W.; Connolly, James L.; Fujimoto, James G.

    2010-02-01

    Excisional biopsy is the current gold standard for disease diagnosis; however, it requires a relatively long processing time and it may also suffer from unacceptable false negative rates due to sampling errors. Optical coherence tomography (OCT) is a promising imaging technique that provide real-time, high resolution and three-dimensional (3D) images of tissue morphology. Optical coherence microscopy (OCM) is an extension of OCT, combining both the coherence gating and the confocal gating techniques. OCM imaging achieves cellular resolution with deeper imaging depth compared to confocal microscopy. An integrated OCT/OCM imaging system can provide co-registered multiscale imaging of tissue morphology. 3D-OCT provides architectural information with a large field of view and can be used to find regions of interest; while OCM provides high magnification to enable cellular imaging. The integrated OCT/OCM system has an axial resolution of <4um and transverse resolutions of 14um and <2um for OCT and OCM, respectively. In this study, a wide range of human pathologic specimens, including colon (58), thyroid (43), breast (34), and kidney (19), were imaged with OCT and OCM within 2 to 6 hours after excision. The images were compared with H & E histology to identify characteristic features useful for disease diagnosis. The feasibility of visualizing human pathology using integrated OCT/OCM was demonstrated in the pathology laboratory settings.

  1. Scanning Tunneling Optical Resonance Microscopy Developed

    NASA Technical Reports Server (NTRS)

    Bailey, Sheila G.; Raffaelle, Ryne P.; Lau, Janis E.; Jenkins, Phillip P.; Castro, Stephanie L.; Tin, Padetha; Wilt, David M.; Pal, Anna Maria; Fahey, Stephen D.

    2004-01-01

    The ability to determine the in situ optoelectronic properties of semiconductor materials has become especially important as the size of device architectures has decreased and the development of complex microsystems has increased. Scanning Tunneling Optical Resonance Microscopy, or STORM, can interrogate the optical bandgap as a function of its position within a semiconductor micro-structure. This technique uses a tunable solidstate titanium-sapphire laser whose output is "chopped" using a spatial light modulator and is coupled by a fiber-optic connector to a scanning tunneling microscope in order to illuminate the tip-sample junction. The photoenhanced portion of the tunneling current is spectroscopically measured using a lock-in technique. The capabilities of this technique were verified using semiconductor microstructure calibration standards that were grown by organometallic vapor-phase epitaxy. Bandgaps characterized by STORM measurements were found to be in good agreement with the bulk values determined by transmission spectroscopy and photoluminescence and with the theoretical values that were based on x-ray diffraction results.

  2. Analyzing Structure and Function of Vascularization in Engineered Bone Tissue by Video-Rate Intravital Microscopy and 3D Image Processing.

    PubMed

    Pang, Yonggang; Tsigkou, Olga; Spencer, Joel A; Lin, Charles P; Neville, Craig; Grottkau, Brian

    2015-10-01

    Vascularization is a key challenge in tissue engineering. Three-dimensional structure and microcirculation are two fundamental parameters for evaluating vascularization. Microscopic techniques with cellular level resolution, fast continuous observation, and robust 3D postimage processing are essential for evaluation, but have not been applied previously because of technical difficulties. In this study, we report novel video-rate confocal microscopy and 3D postimage processing techniques to accomplish this goal. In an immune-deficient mouse model, vascularized bone tissue was successfully engineered using human bone marrow mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs) in a poly (D,L-lactide-co-glycolide) (PLGA) scaffold. Video-rate (30 FPS) intravital confocal microscopy was applied in vitro and in vivo to visualize the vascular structure in the engineered bone and the microcirculation of the blood cells. Postimage processing was applied to perform 3D image reconstruction, by analyzing microvascular networks and calculating blood cell viscosity. The 3D volume reconstructed images show that the hMSCs served as pericytes stabilizing the microvascular network formed by HUVECs. Using orthogonal imaging reconstruction and transparency adjustment, both the vessel structure and blood cells within the vessel lumen were visualized. Network length, network intersections, and intersection densities were successfully computed using our custom-developed software. Viscosity analysis of the blood cells provided functional evaluation of the microcirculation. These results show that by 8 weeks, the blood vessels in peripheral areas function quite similarly to the host vessels. However, the viscosity drops about fourfold where it is only 0.8 mm away from the host. In summary, we developed novel techniques combining intravital microscopy and 3D image processing to analyze the vascularization in engineered bone. These techniques have broad

  3. 3D papillary image capturing by the stereo fundus camera system for clinical diagnosis on retina and optic nerve

    NASA Astrophysics Data System (ADS)

    Motta, Danilo A.; Serillo, André; de Matos, Luciana; Yasuoka, Fatima M. M.; Bagnato, Vanderlei S.; Carvalho, Luis A. V.

    2014-03-01

    Glaucoma is the second main cause of the blindness in the world and there is a tendency to increase this number due to the lifetime expectation raise of the population. Glaucoma is related to the eye conditions, which leads the damage to the optic nerve. This nerve carries visual information from eye to brain, then, if it has damage, it compromises the visual quality of the patient. In the majority cases the damage of the optic nerve is irreversible and it happens due to increase of intraocular pressure. One of main challenge for the diagnosis is to find out this disease, because any symptoms are not present in the initial stage. When is detected, it is already in the advanced stage. Currently the evaluation of the optic disc is made by sophisticated fundus camera, which is inaccessible for the majority of Brazilian population. The purpose of this project is to develop a specific fundus camera without fluorescein angiography and red-free system to accomplish 3D image of optic disc region. The innovation is the new simplified design of a stereo-optical system, in order to make capable the 3D image capture and in the same time quantitative measurements of excavation and topography of optic nerve; something the traditional fundus cameras do not do. The dedicated hardware and software is developed for this ophthalmic instrument, in order to permit quick capture and print of high resolution 3D image and videos of optic disc region (20° field-of-view) in the mydriatic and nonmydriatic mode.

  4. Monitoring adipose-derived stem cells within 3D carrier by combined dielectric spectroscopy and spectral domain optical coherence topography

    NASA Astrophysics Data System (ADS)

    Bagnaninchi, P. O.

    2010-02-01

    Monitoring non-invasively the cellular events in three dimensional carriers is a major challenge for tissue engineering and regenerative medicine that prevents time-lapsed studies over large population of sample. The potential of optical coherence tomography has been demonstrated to assess tissue formation within porous matrices. In this study we explore the combination of dielectric spectroscopy (DS) and spectral domain optical coherence tomography (SDOCT) to quality assess ADSCs loaded in three dimensional carriers. A SDOCT (930nm, FWHM 90nm) was combined to an open ended coaxial probe connected to material analyser, and broadband measurements between 20MHz and 1GHz were synchronized with Labview. Both ADSCs maintained in undifferentiated state within 3D carrier and induced towards osteoblasts were monitored with this multimodality technique and their DS spectra were acquired at high cell concentration simultaneously to 3D imaging. This multimodality technique will be instrumental to assess non-invasively cell loaded carriers for cell therapy.

  5. Fast optical sectioning obtained by structured illumination microscopy using a digital mirror device

    NASA Astrophysics Data System (ADS)

    Xu, Dongli; Jiang, Tao; Li, Anan; Hu, Bihe; Feng, Zhao; Gong, Hui; Zeng, Shaoqun; Luo, Qingming

    2013-06-01

    High-throughput optical imaging is critical to obtain large-scale neural connectivity information of brain in neuroscience. Using a digital mirror device and a scientific complementary metal-oxide semiconductor camera, we report a significant speed improvement of structured illumination microscopy (SIM), which produces a maximum SIM net frame rate of 133 Hz. We perform three-dimensional (3-D) imaging of mouse brain slices at diffraction-limited resolution and demonstrate the fast 3-D imaging capability to a large sample with an imaging rate of 6.9 pixel/s of our system, an order of magnitude faster than previously reported.

  6. Flattop beam illumination for 3D imaging ladar with simple optical devices in the wide distance range

    NASA Astrophysics Data System (ADS)

    Tsuji, Hidenobu; Nakano, Takayuki; Matsumoto, Yoshihiro; Kameyama, Shumpei

    2016-04-01

    We have developed an illumination optical system for 3D imaging ladar (laser detection and ranging) which forms flattop beam shape by transformation of the Gaussian beam in the wide distance range. The illumination is achieved by beam division and recombination using a prism and a negative powered lens. The optimum condition of the transformation by the optical system is derived. It is confirmed that the flattop distribution can be formed in the wide range of the propagation distance from 1 to 1000 m. The experimental result with the prototype is in good agreement with the calculation result.

  7. Optical coherence photoacoustic microscopy: accomplishing optical coherence tomography and photoacoustic microscopy with a single light source

    PubMed Central

    Zhang, Xiangyang; Zhang, Hao F.

    2012-01-01

    Abstract. We developed optical coherence photoacoustic microscopy (OC-PAM) to demonstrate that the functions of optical coherence tomography (OCT) and photoacoustic microscopy (PAM) can be achieved simultaneously by using a single illuminating light source. We used a pulsed broadband laser centered at 580 nm and detected the absorbed photons through photoacoustic detection and the back-scattered photons with an interferometer. In OC-PAM, each laser pulse generates both one OCT A-line and one PAM A-line simultaneously; as a result, the two imaging modalities are intrinsically co-registered in the lateral directions. In vivo images of the mouse ear were acquired to demonstrate the capabilities of OC-PAM. PMID:22502553

  8. Optical fibre sensing in metals by embedment in 3D printed metallic structures

    NASA Astrophysics Data System (ADS)

    Maier, R. R. J.; Havermann, D.; Schneller, O.; Mathew, J.; Polyzos, D.; MacPherson, W. N.; Hand, D. P.

    2014-05-01

    Additive manufacturing or 3D printing of structural components in metals has potential to revolutionise the manufacturing industry. Embedded sensing in such structures opens a route towards SMART metals, providing added functionality, intelligence and enhanced performance in many components. Such embedded sensors would be capable of operating at extremely high temperatures by utilizing regenerated fibre Bragg gratings and in-fibre Fabry-Perot cavities.

  9. Towards a noninvasive intracranial tumor irradiation using 3d optical imaging and multimodal data registration.

    PubMed

    Posada, R; Daul, Ch; Wolf, D; Aletti, P

    2007-01-01

    Conformal radiotherapy (CRT) results in high-precision tumor volume irradiation. In fractioned radiotherapy (FRT), lesions are irradiated in several sessions so that healthy neighbouring tissues are better preserved than when treatment is carried out in one fraction. In the case of intracranial tumors, classical methods of patient positioning in the irradiation machine coordinate system are invasive and only allow for CRT in one irradiation session. This contribution presents a noninvasive positioning method representing a first step towards the combination of CRT and FRT. The 3D data used for the positioning is point clouds spread over the patient's head (CT-data usually acquired during treatment) and points distributed over the patient's face which are acquired with a structured light sensor fixed in the therapy room. The geometrical transformation linking the coordinate systems of the diagnosis device (CT-modality) and the 3D sensor of the therapy room (visible light modality) is obtained by registering the surfaces represented by the two 3D point sets. The geometrical relationship between the coordinate systems of the 3D sensor and the irradiation machine is given by a calibration of the sensor position in the therapy room. The global transformation, computed with the two previous transformations, is sufficient to predict the tumor position in the irradiation machine coordinate system with only the corresponding position in the CT-coordinate system. Results obtained for a phantom show that the mean positioning error of tumors on the treatment machine isocentre is 0.4 mm. Tests performed with human data proved that the registration algorithm is accurate (0.1 mm mean distance between homologous points) and robust even for facial expression changes.

  10. Pre-Processing of Point-Data from Contact and Optical 3D Digitization Sensors

    PubMed Central

    Budak, Igor; Vukelić, Djordje; Bračun, Drago; Hodolič, Janko; Soković, Mirko

    2012-01-01

    Contemporary 3D digitization systems employed by reverse engineering (RE) feature ever-growing scanning speeds with the ability to generate large quantity of points in a unit of time. Although advantageous for the quality and efficiency of RE modelling, the huge number of point datas can turn into a serious practical problem, later on, when the CAD model is generated. In addition, 3D digitization processes are very often plagued by measuring errors, which can be attributed to the very nature of measuring systems, various characteristics of the digitized objects and subjective errors by the operator, which also contribute to problems in the CAD model generation process. This paper presents an integral system for the pre-processing of point data, i.e., filtering, smoothing and reduction, based on a cross-sectional RE approach. In the course of the proposed system development, major emphasis was placed on the module for point data reduction, which was designed according to a novel approach with integrated deviation analysis and fuzzy logic reasoning. The developed system was verified through its application on three case studies, on point data from objects of versatile geometries obtained by contact and laser 3D digitization systems. The obtained results demonstrate the effectiveness of the system. PMID:22368513

  11. Writing of 3D optical integrated circuits with ultrashort laser pulses in the presence of strong spherical aberration

    NASA Astrophysics Data System (ADS)

    Bukharin, M. A.; Skryabin, N. N.; Khudyakov, D. V.; Vartapetov, S. K.

    2016-09-01

    A novel technique was proposed for 3D femtosecond writing of waveguides and optical integrated circuits in the presence of strong spherical aberration, caused by inscription at significantly different depth under the surface of optical glasses and crystals. Strong negative effect of spherical aberration and related asymmetry of created structures was reduced due to transition to the cumulative thermal regime of femtosecond interaction with the material. The differences in the influence of spherical aberration effect in a broad depth range (larger than 200 µm) was compensated by dynamic adjustment of laser pulse energy during the process of waveguides recording. The presented approach has been experimentally implemented in fused silica. Obtained results can be used in production of a broad class of femtosecond written three-dimensional integrated optical systems, inscripted at non-optimal (for focusing lens) optical depth or in significantly extended range of depths.

  12. Lab on chip optical imaging of biological sample by quantitative phase microscopy

    NASA Astrophysics Data System (ADS)

    Memmolo, P.; Miccio, L.; Merola, F.; Gennari, O.; Mugnano, M.; Netti, P. A.; Ferraro, P.

    2015-03-01

    Quantitative imaging and three dimensional (3D) morphometric analysis of flowing and not-adherent cells is an important aspect for diagnostic purposes at Lab on Chip scale. Diagnostics tools need to be quantitative, label-free and, as much as possible, accurate. In recent years digital holography (DH) has been improved to be considered as suitable diagnostic method in several research field. In this paper we demonstrate that DH can be used for retrieving 3D morphometric data for sorting and diagnosis aims. Several techniques exist for 3D morphological study as optical coherent tomography and confocal microscopy, but they are not the best choice in case of dynamic events as flowing samples. Recently, a DH approach, based on shape from silhouette algorithm (SFS), has been developed for 3D shape display and calculation of cells biovolume. Such approach, adopted in combination with holographic optical tweezers (HOT) was successfully applied to cells with convex shape. Unfortunately, it's limited to cells with convex surface as sperm cells or diatoms. Here, we demonstrate an improvement of such procedure. By decoupling thickness information from refractive index ones and combining this with SFS analysis, 3D shape of concave cells is obtained. Specifically, the topography contour map is computed and used to adjust the 3D shape retrieved by the SFS algorithm. We prove the new procedure for healthy red blood cells having a concave surface in their central region. Experimental results are compared with theoretical model.

  13. New method for detection of complex 3D fracture motion - Verification of an optical motion analysis system for biomechanical studies

    PubMed Central

    2012-01-01

    Background Fracture-healing depends on interfragmentary motion. For improved osteosynthesis and fracture-healing, the micromotion between fracture fragments is undergoing intensive research. The detection of 3D micromotions at the fracture gap still presents a challenge for conventional tactile measurement systems. Optical measurement systems may be easier to use than conventional systems, but, as yet, cannot guarantee accuracy. The purpose of this study was to validate the optical measurement system PONTOS 5M for use in biomechanical research, including measurement of micromotion. Methods A standardized transverse fracture model was created to detect interfragmentary motions under axial loadings of up to 200 N. Measurements were performed using the optical measurement system and compared with a conventional high-accuracy tactile system consisting of 3 standard digital dial indicators (1 μm resolution; 5 μm error limit). Results We found that the deviation in the mean average motion detection between the systems was at most 5.3 μm, indicating that detection of micromotion was possible with the optical measurement system. Furthermore, we could show two considerable advantages while using the optical measurement system. Only with the optical system interfragmentary motion could be analyzed directly at the fracture gap. Furthermore, the calibration of the optical system could be performed faster, safer and easier than that of the tactile system. Conclusion The PONTOS 5 M optical measurement system appears to be a favorable alternative to previously used tactile measurement systems for biomechanical applications. Easy handling, combined with a high accuracy for 3D detection of micromotions (≤ 5 μm), suggests the likelihood of high user acceptance. This study was performed in the context of the deployment of a new implant (dynamic locking screw; Synthes, Oberdorf, Switzerland). PMID:22405047

  14. Correlation between Optic Nerve Parameters Obtained Using 3D Nonmydriatic Retinal Camera and Optical Coherence Tomography: Interobserver Agreement on the Disc Damage Likelihood Scale.

    PubMed

    Han, Jae Wook; Cho, Soon Young; Kang, Kui Dong

    2014-01-01

    Purpose. To compare stereometric parameters obtained by three-dimensional (3D) optic disc photography and optical coherence tomography (OCT) and assess interobserver agreement on the disc damage likelihood scale (DDLS). Methods. This retrospective study included 190 eyes from 190 patients classified as normal, glaucoma suspect, or glaucomatous. Residents at different levels of training completed the DDLS for each patient before and after attending a training module. 3D optic disc photography and OCT were performed on each eye, and correlations between the DDLS and various parameters obtained by each device were calculated. Results. We found moderate agreement (weighted kappa value, 0.59 ± 0.03) between DDLS scores obtained by 3D optic disc photography and the glaucoma specialist. The weighted kappa values for agreement and interobserver concordance increased among residents after the training module. Interobserver concordance was the poorest at DDLS stages 5 and 6. The DDLS scored by the glaucoma specialist had the highest predictability value (0.941). Conclusions. The DDLS obtained by 3D optic disc photography is a useful diagnostic tool for glaucoma. A supervised teaching program increased trainee interobserver agreement on the DDLS. DDLS stages 5 and 6 showed the poorest interobserver agreement, suggesting that caution is required when recording these stages.

  15. Correlation between Optic Nerve Parameters Obtained Using 3D Nonmydriatic Retinal Camera and Optical Coherence Tomography: Interobserver Agreement on the Disc Damage Likelihood Scale.

    PubMed

    Han, Jae Wook; Cho, Soon Young; Kang, Kui Dong

    2014-01-01

    Purpose. To compare stereometric parameters obtained by three-dimensional (3D) optic disc photography and optical coherence tomography (OCT) and assess interobserver agreement on the disc damage likelihood scale (DDLS). Methods. This retrospective study included 190 eyes from 190 patients classified as normal, glaucoma suspect, or glaucomatous. Residents at different levels of training completed the DDLS for each patient before and after attending a training module. 3D optic disc photography and OCT were performed on each eye, and correlations between the DDLS and various parameters obtained by each device were calculated. Results. We found moderate agreement (weighted kappa value, 0.59 ± 0.03) between DDLS scores obtained by 3D optic disc photography and the glaucoma specialist. The weighted kappa values for agreement and interobserver concordance increased among residents after the training module. Interobserver concordance was the poorest at DDLS stages 5 and 6. The DDLS scored by the glaucoma specialist had the highest predictability value (0.941). Conclusions. The DDLS obtained by 3D optic disc photography is a useful diagnostic tool for glaucoma. A supervised teaching program increased trainee interobserver agreement on the DDLS. DDLS stages 5 and 6 showed the poorest interobserver agreement, suggesting that caution is required when recording these stages. PMID:24804081

  16. The 3D manipulation of a microsphere for nano-CMM probe using single fiber optical trapping

    NASA Astrophysics Data System (ADS)

    Eom, Sang In; Takaya, Yasuhiro; Miyoshi, Takashi; Hayashi, Terutake

    2006-08-01

    We suggest a novel 3-D probing technique for measuring micro parts which have high aspect ratio such as a groove or a deep-hole. This technique uses the optical force exerted on a dielectric microsphere at the tip of optical fiber so called the fiber optical trapping. A microsphere is trapped at the tip of an optical fiber which has micrometer size of the diameter and sub micrometer size of the tapered tip. The optical source is a Nd:YAG laser with the wavelength of 1064 nm. A fiber optical trapping system is quite similar to a conventional CMM. The roll of a microsphere is the same as the probe sphere of a conventional CMM and an optical fiber works as a stylus shaft. The micro optical fiber part is thinner than the diameter of the microsphere and longer than the depth of shapes such as deep holes and grooves, which enable to make an approach to a steep angle surface of a work piece with high aspect ratio.

  17. WebTOP: A 3D Interactive System for Teaching and Learning Optics

    ERIC Educational Resources Information Center

    Mzoughi, Taha; Herring, S. Davis; Foley, John T.; Morris, Matthew J.; Gilbert, Peter J.

    2007-01-01

    WebTOP is a three-dimensional, Web-based, interactive computer graphics system that helps instructors teach and students learn about waves and optics. Current subject areas include waves, geometrical optics, reflection and refraction, polarization, interference, diffraction, lasers, and scattering. Some of the topics covered are suited for…

  18. 3D monitoring and quality control using intraoral optical camera systems.

    PubMed

    Mehl, A; Koch, R; Zaruba, M; Ender, A

    2013-01-01

    The quality of intraoral scanning systems is steadily improving, and they are becoming easier and more reliable to operate. This opens up possibilities for routine clinical applications. A special aspect is that overlaying (superimposing) situations recorded at different times facilitates an accurate three-dimensional difference analysis. Such difference analyses can also be used to advantage in other areas of dentistry where target/actual comparisons are required. This article presents potential indications using a newly developed software, explaining the functionality of the evaluation process and the prerequisites and limitations of 3D monitoring.

  19. Single qubit gates on neutral atoms in a 3d Optical lattice

    NASA Astrophysics Data System (ADS)

    Kumar, Aishwarya; Wang, Yang; Zhang, Xianli; Corcovilos, Theodore A.; Weiss, David S.

    2015-05-01

    Neutral atoms are especially promising candidates for quantum computing because of their inherent scalability. To realize this scalability requires being able to manipulate the quantum information at target qubits with high fidelity and low crosstalk. We will present two single qubit gate addressing protocols. We have experimentally applied them both to targeted sites in a 5 × 5 × 5 3D array. The two distinct approaches both use crossed MEMS-mirror directed addressing beams along with microwave pulses to target atoms at single sites, while having minimal impact on the quantum information at non-target sites. Supported by DARPA, QUEST and ARO.

  20. [Morton's disease: optic and electron microscopy observations].

    PubMed

    De Palma, L; Tulli, A

    1991-01-01

    The authors performed an optic and electron-microscope investigation above the common digital nerve of the foot, whose fragments had been surgically removed from patients suffering from "Morton metatarsalgia" (neuroma). Histological sections were taken from pre-stenotic swelling in patients with clinical symptoms persisting for one year; perineural thickening without evidence of fibroblastic proliferation could be demonstrated, together with an intraneural deposition of an amorphous substance. In other patients suffering from Morton's disease for a longer time, a more pronounced epineural thickening in the pre-stenotic zone could be shown, with partial replacement of nerve fibers by amorphous substance. In the same patients endoneural fibrositis was seen at the level of the stenosis. Electron-microscopy in patients after one year showed an increase in collagenous endoneural fibers and microfibrils. These histopathological findings suggest a compressive mechanism in the pathogenesis of the damage to the common interdigital nerve in Morton's disease, caused by the extrinsic anatomical structures surrounding the nerve. The so-called "neuroma" can be identified with the pre-stenotic swelling.

  1. Segmentation of whole cells and cell nuclei from 3-D optical microscope images using dynamic programming.

    PubMed

    McCullough, D P; Gudla, P R; Harris, B S; Collins, J A; Meaburn, K J; Nakaya, M A; Yamaguchi, T P; Misteli, T; Lockett, S J

    2008-05-01

    Communications between cells in large part drive tissue development and function, as well as disease-related processes such as tumorigenesis. Understanding the mechanistic bases of these processes necessitates quantifying specific molecules in adjacent cells or cell nuclei of intact tissue. However, a major restriction on such analyses is the lack of an efficient method that correctly segments each object (cell or nucleus) from 3-D images of an intact tissue specimen. We report a highly reliable and accurate semi-automatic algorithmic method for segmenting fluorescence-labeled cells or nuclei from 3-D tissue images. Segmentation begins with semi-automatic, 2-D object delineation in a user-selected plane, using dynamic programming (DP) to locate the border with an accumulated intensity per unit length greater that any other possible border around the same object. Then the two surfaces of the object in planes above and below the selected plane are found using an algorithm that combines DP and combinatorial searching. Following segmentation, any perceived errors can be interactively corrected. Segmentation accuracy is not significantly affected by intermittent labeling of object surfaces, diffuse surfaces, or spurious signals away from surfaces. The unique strength of the segmentation method was demonstrated on a variety of biological tissue samples where all cells, including irregularly shaped cells, were accurately segmented based on visual inspection.

  2. 3D optical two-mirror scanner with focus-tunable lens.

    PubMed

    Pokorny, Petr; Miks, Antonin

    2015-08-01

    The paper presents formulas for a ray tracing in the optical system of two-mirror optical scanner with a focus-tunable lens. Furthermore, equations for the calculation of focal length which ensure focusing of a beam in the desired point in a detection plane are derived. The uncertainty description of such focal length follows as well. The chosen vector approach is general; therefore, the application of formulas in various configurations of the optical systems is possible. In the example situation, the authors derived formulas for mirrors' rotations and the focal length depending on the position of the point in the detection plane. PMID:26368115

  3. FT3D: three-dimensional Fourier analysis on small Unix workstations for electron microscopy and tomographic studies.

    PubMed

    Lanzavecchia, S; Bellon, P L; Tosoni, L

    1993-12-01

    FT3D is a self-contained package of tools for three-dimensional Fourier analysis, written in the C language for Unix workstations. It can evaluate direct transforms of three-dimensional real functions, inverse transforms, auto- and cross-correlations and spectra. The library has been developed to support three-dimensional reconstructions of biological structures from projections obtained in the electron microscope. This paper discusses some features of the library, which has been implemented in such a way as to profit from the resources of modern workstations. A table of elapsed times for jobs of different dimensions with different RAM buffers is reported for the particular hardware used in the authors' laboratory.

  4. Feasibility study of a single-shot 3D electron bunch shape monitor with an electro-optic sampling technique

    NASA Astrophysics Data System (ADS)

    Okayasu, Yuichi; Tomizawa, Hiromitsu; Matsubara, Shinichi; Kumagai, Noritaka; Maekawa, Akira; Uesaka, Mitsuru; Ishikawa, Tetsuya

    2013-05-01

    We developed a three-dimensional electron bunch charge distribution (3D-BCD) monitor with single-shot detection, and a spectral decoding based electro-optic (EO) sampling technique for a nondestructive monitor enables real-time reconstruction of the three-dimensional distribution of a bunch charge. We realized three goals by simultaneously probing a number of Pockels EO crystals that surround the electron beam axis with hollow and radial polarized laser pulses. First, we performed a feasibility test as a simple case of a 3D-BCD monitor probing two ZnTe crystals as EO detectors installed on the opposite angle to the electron beam axis and confirmed that we simultaneously obtained both EO signals. Since the adopted hollow probe laser pulse is not only radially polarized but also temporally shifted azimuthally, some disorders in the radial polarization distribution of such a laser pulse were numerically analyzed with a plane-wave expansion method. Based on the above investigations, the 3D-BCD monitor is feasible both in experimental and numerical estimations. Furthermore, we previously developed a femtosecond response organic crystal as a Pockels EO detector and a broadband probe laser (≥350nm in FWHM); the 3D-BCD monitor realizes 30- to 40-fs (FWHM) temporal resolution. Eventually, the monitor is expected to be equipped in such advanced accelerators as XFEL to measure and adjust the electron bunch charge distribution in real time. The 3D-BCD measurement works as a critical tool to provide feedback to seeded FELs.

  5. Operational Retrieval of aerosol optical depth over Indian subcontinent and Indian Ocean using INSAT-3D/Imager product validation

    NASA Astrophysics Data System (ADS)

    Mishra, M. K.; Rastogi, G.; Chauhan, P.

    2014-11-01

    Aerosol optical depth (AOD) over Indian subcontinent and Indian Ocean region is derived operationally for the first time from the geostationary earth orbit (GEO) satellite INSAT-3D Imager data at 0.65 μm wavelength. Single visible channel algorithm based on clear sky composites gives larger retrieval error in AOD than other multiple channel algorithms due to errors in estimating surface reflectance and atmospheric property. However, since MIR channel signal is insensitive to the presence of most aerosols, therefore in present study, AOD retrieval algorithm employs both visible (centred at 0.65 μm) and mid-infrared (MIR) band (centred at 3.9 μm) measurements, and allows us to monitor transport of aerosols at higher temporal resolution. Comparisons made between INSAT-3D derived AOD (τI) and MODIS derived AOD (τM) co-located in space (at 1° resolution) and time during January, February and March (JFM) 2014 encompasses 1165, 1052 and 900 pixels, respectively. Good agreement found between τI and τM during JFM 2014 with linear correlation coefficients (R) of 0.87, 0.81 and 0.76, respectively. The extensive validation made during JFM 2014 encompasses 215 co-located AOD in space and time derived by INSAT 3D (τI) and 10 sun-photometers (τA) that includes 9 AERONET (Aerosol Robotic Network) and 1 handheld sun-photometer site. INSAT-3D derived AOD i.e. τI, is found within the retrieval errors of τI = ±0.07 ±0.15τA with linear correlation coefficient (R) of 0.90 and root mean square error equal (RMSE) to 0.06. Present work shows that INSAT-3D aerosol products can be used quantitatively in many applications with caution for possible residual clouds, snow/ice, and water contamination.

  6. Treating benign optic nerve tumors with a 3-D conformal plan

    SciTech Connect

    Millunchick, Cheryl Hope

    2013-07-01

    A 68 year old male patient presented for radiation therapy for treatment of a benign tumor, a glioma of his left optic nerve. The radiation oncologist intended to prescribe 52.2 Gy to the planning target volume, while maintaining a maximum of 54 Gy to the optic nerves and the optic chiasm and a maximum of 40–45 Gy to the globes in order to minimize the possibility of damaging the optic system, which is especially important as this is a benign tumor. The dosimetrist devised a conformal non-coplanar three-dimensional plan with a slightly weighted forward planning component. This plan was created in approximately 15 minutes after the critical organs and the targets were delineated and resulted in an extremely conformal and homogenous plan, treating the target while sparing the nearby critical structures. This approach can also be extended to other tumors in the brain - benign or malignant.

  7. Full-color holographic 3D imaging system using color optical scanning holography

    NASA Astrophysics Data System (ADS)

    Kim, Hayan; Kim, You Seok; Kim, Taegeun

    2016-06-01

    We propose a full-color holographic three-dimensional imaging system that composes a recording stage, a transmission and processing stage and reconstruction stage. In recording stage, color optical scanning holography (OSH) records the complex RGB holograms of an object. In transmission and processing stage, the recorded complex RGB holograms are transmitted to the reconstruction stage after conversion to off-axis RGB holograms. In reconstruction stage, the off-axis RGB holograms are reconstructed optically.

  8. Cost-effective method of manufacturing a 3D MEMS optical switch

    NASA Astrophysics Data System (ADS)

    Carr, Emily; Zhang, Ping; Keebaugh, Doug; Chau, Kelvin

    2009-02-01

    growth of data and video transport networks. All-optical switching eliminates the need for optical-electrical conversion offering the ability to switch optical signals transparently: independent of data rates, formats and wavelength. It also provides network operators much needed automation capabilities to create, monitor and protect optical light paths. To further accelerate the market penetration, it is necessary to identify a path to reduce the manufacturing cost significantly as well as enhance the overall system performance, uniformity and reliability. Currently, most MEMS optical switches are assembled through die level flip-chip bonding with either epoxies or solder bumps. This is due to the alignment accuracy requirements of the switch assembly, defect matching of individual die, and cost of the individual components. In this paper, a wafer level assembly approach is reported based on silicon fusion bonding which aims to reduce the packaging time, defect count and cost through volume production. This approach is successfully demonstrated by the integration of two 6-inch wafers: a mirror array wafer and a "snap-guard" wafer, which provides a mechanical structure on top of the micromirror to prevent electrostatic snap-down. The direct silicon-to-silicon bond eliminates the CTEmismatch and stress issues caused by non-silicon bonding agents. Results from a completed integrated switch assembly will be presented, which demonstrates the reliability and uniformity of some key parameters of this MEMS optical switch.

  9. LES prediction and analysis of the aero-optical environment around a 3-D turret

    NASA Astrophysics Data System (ADS)

    Mathews, Edwin; Wang, Kan; Wang, Meng; Jumper, Eric

    2015-11-01

    Using wall-modeled large-eddy simulation, a Mach 0.4 flow over a hemisphere-on-cylinder turret at the experimental Reynolds number of ReD = 2 . 3 ×106 is simulated to study the aero-optical distortions caused by turbulent density fluctuations. The optical distortions are ca