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Sample records for 3d optical microscopy

  1. 3-D Optical Interference Microscopy at the Lateral Resolution

    NASA Astrophysics Data System (ADS)

    Lehmann, Peter; Niehues, Jan; Tereschenko, Stanislav

    2014-10-01

    For applications in micro- and nanotechnologies the lateral resolution of optical 3-D microscopes becomes an issue of increasing relevance. However, lateral resolution of 3-D microscopes is hard to define in a satisfying way. Therefore, we first study the measurement capabilities of a highly resolving white-light interference (WLI) microscope close to the limit of lateral resolution. Results of measurements and simulations demonstrate that better lateral resolution seems to be achievable based on the envelope evaluation of a WLI signal. Unfortunately, close to the lateral resolution limit errors in the measured amplitude of micro-structures appear. On the other hand, results of interferometric phase evaluation seem to be strongly low-pass filtered in this case. Furthermore, the instrument transfer characteristics and the lateral resolution capabilities of WLI instruments are also affected by polarization. TM polarized light is less sensitive to edge diffraction and thus systematic errors can be avoided. However, apart from ghost steps due to fringe order errors, the results of phase evaluation seem to be closer to the real surface topography if TE polarized light is used. The lateral resolution can be further improved by combining WLI and structured illumination microscopy. Since the measured height of rectangular profiles close to the lateral resolution limit is generally too small compared to the real height, we introduce a method based on phase evaluation which characterizes the heights of barely laterally resolved rectangular gratings correctly.

  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. 3D defect detection using optical wide-field microscopy

    NASA Astrophysics Data System (ADS)

    Tympel, Volker; Schaaf, Marko; Srocka, Bernd

    2007-06-01

    We report a method to detect signed differences in two similar data sets representing 3-dimensional intensity profiles recorded by optical wide-field microscopes. The signed differences describe missing or unexpected intensity values, defined as defects. In technical applications like wafer and mask inspection, data sets often represent surfaces. The reported method is able to describe the size and position especially in relation to the neighboring surface and is called Three-Dimension-Aberration (TDA)-Technology. To increase the tool performance and to handle different sizes of defects a scaled bottom-up method is implemented and started with high reduced data sets for the search of large defects. Each analysis contains three steps. The first step is a correlation to calculate the displacement vector between the similar data sets. In the second step a new data set is created. The new data set consists of intensity differences. Extreme values in the data set represent the position of defects. By the use of linear and non-linear filters the stability of detection can be improved. If all differences are below a threshold the bottom-up method starts with the next larger scaled data set. In the other case it is assumed that the defect is detected and step three starts with the detection of the convex hull of the defect and the search of the neighboring surface. As a result the defect is described by a parameter set including the relative position. Because of the layered structure of the data set and the bottom-up technique the method is suitable for multi-core processor architectures.

  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-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

  8. 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

  9. 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

  10. Analytic 3D Imaging of Mammalian Nucleus at Nanoscale Using Coherent X-Rays and Optical Fluorescence Microscopy

    PubMed Central

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

    2014-01-01

    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. PMID:25185543

  11. Multicolor 3D super-resolution imaging by quantum dot stochastic optical reconstruction microscopy.

    PubMed

    Xu, Jianquan; Tehrani, Kayvan F; Kner, Peter

    2015-03-24

    We demonstrate multicolor three-dimensional super-resolution imaging with quantum dots (QSTORM). By combining quantum dot asynchronous spectral blueing with stochastic optical reconstruction microscopy and adaptive optics, we achieve three-dimensional imaging with 24 nm lateral and 37 nm axial resolution. By pairing two short-pass filters with two appropriate quantum dots, we are able to image single blueing quantum dots on two channels simultaneously, enabling multicolor imaging with high photon counts. PMID:25703291

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

    NASA Astrophysics Data System (ADS)

    Casal, A.; Cerrato, R.; Mateo, M. P.; Nicolas, G.

    2016-06-01

    A low-cost optical microscope was developed and coupled to an irradiation system in order to study the induced effects on material during a multipulse regime by an in situ visual inspection of the surface, in particular of the spot generated at different pulses. In the case of laser ablation, a reconstruction of the crater in 3D was made from the images of the sample surface taken during the irradiation process, and the subsequent profiles of ablated material were extracted. The implementation of this homemade optical device gives an added value to the irradiation system, providing information about morphology evolution of irradiated area when successive pulses are applied. In particular, the determination of ablation rates in real time can be especially useful for a better understanding and controlling of the ablation process in applications where removal of material is involved, such as laser cleaning and in-depth characterization of multilayered samples and diffusion processes. The validation of the developed microscope was made by a comparison with a commercial confocal microscope configured for the characterization of materials where similar results of crater depth and diameter were obtained for both systems.

  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

    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

  17. 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

  18. 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

  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. 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.

  1. 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

  2. 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.

  3. Holography, tomography and 3D microscopy as linear filtering operations

    NASA Astrophysics Data System (ADS)

    Coupland, J. M.; Lobera, J.

    2008-07-01

    In this paper, we characterize 3D optical imaging techniques as 3D linear shift-invariant filtering operations. From the Helmholtz equation that is the basis of scalar diffraction theory, we show that the scattered field, or indeed a holographic reconstruction of this field, can be considered to be the result of a linear filtering operation applied to a source distribution. We note that if the scattering is weak, the source distribution is independent of the scattered field and a holographic reconstruction (or in fact any far-field optical imaging system) behaves as a 3D linear shift-invariant filter applied to the refractive index contrast (which effectively defines the object). We go on to consider tomographic techniques that synthesize images from recordings of the scattered field using different illumination conditions. In our analysis, we compare the 3D response of monochromatic optical tomography with the 3D imagery offered by confocal microscopy and scanning white light interferometry (using quasi-monochromatic illumination) and explain the circumstances under which these approaches are equivalent. Finally, we consider the 3D response of polychromatic optical tomography and in particular the response of spectral optical coherence tomography and scanning white light interferometry.

  4. Correlation and Characterization of 3D Morphological Dependent Localized Surface Plasmon Resonance Spectra of Single Silver Nanoparticles Using Dark-field Optical Microscopy and Spectroscopy and AFM

    PubMed Central

    Song, Yujun; Nallathamby, Prakash D.; Huang, Tao; Elsayed-Ali, Hani E.; Xu, Xiao-Hong Nancy

    2009-01-01

    We have developed a new and effective methodology to correlate optical and AFM images of single Ag nanoparticles (NPs), allowing us to study 3D-morphological dependent localized surface plasmon resonance (LSPR) spectra of individual Ag NPs. We fabricated arrays of distinctive microwindows on glass coverslips using photo-lithography method, and created well-isolated individual Ag NPs with a wide variety of shapes and morphologies on the glass coverslips using a modified nanosphere lithography method (NSL). Using distinctive geometries of microwindows, we located individual Ag NPs of interest in their optical and AFM images, enabling us to correlate and characterize the LSPR spectra and 3D morphologies of the same single NPs using dark-field optical microscopy and spectroscopy (DFOMS) and AFM, respectively. We found that LSPR spectra of single Ag NPs, with nearly equal volume [(8.6 ± 0.4) × 103 nm3], cross-section [(2.2 ± 0.2) × 102 nm3], and height (39.6 ± 3.6 nm), highly depend on their shapes, showing the red shift of peak wavelength to 629 nm (quasi trapezoidal cylindrical NP) from that of 506 nm (quasi circular cylindrical NP). LSPR spectra of single Ag NPs simulated using discrete dipole approximation (DDA) agree well with those measured experimentally when their shapes and morphologies can be accuractely described in both methods, but differ when they are not. Furthermore, we found location-dependent LSPR spectra on and around a single NP, offering a unique opportunity to characterize multi-mode plasmonic NPs at nanometer resolution for better understanding their plasmonic optical properties and for rational design of single NP optics. PMID:20190865

  5. Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions

    PubMed Central

    Jun, Sangmi; Zhao, Gongpu; Ning, Jiying; Gibson, Gregory A.; Watkins, Simon C.; Zhang, Peijun

    2013-01-01

    Cryo-electron tomography (cryoET) allows 3D visualization of cellular structures at molecular resolution in a close-to-physiological state1. However, direct visualization of individual viral complexes in their host cellular environment with cryoET is challenging2, due to the infrequent and dynamic nature of viral entry, particularly in the case of HIV-1. While time-lapse live-cell imaging has yielded a great deal of information about many aspects of the life cycle of HIV-13-7, the resolution afforded by live-cell microscopy is limited (~ 200 nm). Our work was aimed at developing a correlation method that permits direct visualization of early events of HIV-1 infection by combining live-cell fluorescent light microscopy, cryo-fluorescent microscopy, and cryoET. In this manner, live-cell and cryo-fluorescent signals can be used to accurately guide the sampling in cryoET. Furthermore, structural information obtained from cryoET can be complemented with the dynamic functional data gained through live-cell imaging of fluorescent labeled target. In this video article, we provide detailed methods and protocols for structural investigation of HIV-1 and host-cell interactions using 3D correlative high-speed live-cell imaging and high-resolution cryoET structural analysis. HeLa cells infected with HIV-1 particles were characterized first by confocal live-cell microscopy, and the region containing the same viral particle was then analyzed by cryo-electron tomography for 3D structural details. The correlation between two sets of imaging data, optical imaging and electron imaging, was achieved using a home-built cryo-fluorescence light microscopy stage. The approach detailed here will be valuable, not only for study of virus-host cell interactions, but also for broader applications in cell biology, such as cell signaling, membrane receptor trafficking, and many other dynamic cellular processes. PMID:23852318

  6. Optically rewritable 3D liquid crystal displays.

    PubMed

    Sun, J; Srivastava, A K; Zhang, W; Wang, L; Chigrinov, V G; Kwok, H S

    2014-11-01

    Optically rewritable liquid crystal display (ORWLCD) is a concept based on the optically addressed bi-stable display that does not need any power to hold the image after being uploaded. Recently, the demand for the 3D image display has increased enormously. Several attempts have been made to achieve 3D image on the ORWLCD, but all of them involve high complexity for image processing on both hardware and software levels. In this Letter, we disclose a concept for the 3D-ORWLCD by dividing the given image in three parts with different optic axis. A quarter-wave plate is placed on the top of the ORWLCD to modify the emerging light from different domains of the image in different manner. Thereafter, Polaroid glasses can be used to visualize the 3D image. The 3D image can be refreshed, on the 3D-ORWLCD, in one-step with proper ORWLCD printer and image processing, and therefore, with easy image refreshing and good image quality, such displays can be applied for many applications viz. 3D bi-stable display, security elements, etc. PMID:25361316

  7. 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

  8. 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

  9. 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.

  10. 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.

  11. 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

  12. 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. PMID:27375935

  13. 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.

  14. 3D Cell Culture Imaging with Digital Holographic Microscopy

    NASA Astrophysics Data System (ADS)

    Dimiduk, Thomas; Nyberg, Kendra; Almeda, Dariela; Koshelva, Ekaterina; McGorty, Ryan; Kaz, David; Gardel, Emily; Auguste, Debra; Manoharan, Vinothan

    2011-03-01

    Cells in higher organisms naturally exist in a three dimensional (3D) structure, a fact sometimes ignored by in vitro biological research. Confinement to a two dimensional culture imposes significant deviations from the native 3D state. One of the biggest obstacles to wider use of 3D cultures is the difficulty of 3D imaging. The confocal microscope, the dominant 3D imaging instrument, is expensive, bulky, and light-intensive; live cells can be observed for only a short time before they suffer photodamage. We present an alternative 3D imaging techinque, digital holographic microscopy, which can capture 3D information with axial resolution better than 2 μm in a 100 μm deep volume. Capturing a 3D image requires only a single camera exposure with a sub-millisecond laser pulse, allowing us to image cell cultures using five orders of magnitude less light energy than with confocal. This can be done with hardware costing ~ 1000. We use the instrument to image growth of MCF7 breast cancer cells and p. pastoras yeast. We acknowledge support from NSF GRFP.

  15. 3D modeling of optically challenging objects.

    PubMed

    Park, Johnny; Kak, Avinash

    2008-01-01

    We present a system for constructing 3D models of real-world objects with optically challenging surfaces. The system utilizes a new range imaging concept called multi-peak range imaging, which stores multiple candidates of range measurements for each point on the object surface. The multiple measurements include the erroneous range data caused by various surface properties that are not ideal for structured-light range sensing. False measurements generated by spurious reflections are eliminated by applying a series of constraint tests. The constraint tests based on local surface and local sensor visibility are applied first to individual range images. The constraint tests based on global consistency of coordinates and visibility are then applied to all range images acquired from different viewpoints. We show the effectiveness of our method by constructing 3D models of five different optically challenging objects. To evaluate the performance of the constraint tests and to examine the effects of the parameters used in the constraint tests, we acquired the ground truth data by painting those objects to suppress the surface-related properties that cause difficulties in range sensing. Experimental results indicate that our method significantly improves upon the traditional methods for constructing reliable 3D models of optically challenging objects. PMID:18192707

  16. Pure optical photoacoustic microscopy

    PubMed Central

    Xie, Zhixing; Chen, Sung-Liang; Ling, Tao; Guo, L. Jay; Carson, Paul L.; Wang, Xueding

    2011-01-01

    The concept of pure optical photoacoustic microscopy(POPAM) was proposed based on optical rastering of a focused excitation beam and optically sensing the photoacoustic signal using a microring resonator fabricated by a nanoimprinting technique. After the refinements of the microring’s working wavelength and in the resonator structure and mold fabrication, an ultrahigh Q factor of 3.0×105 was achieved which provided high sensitivity with a noise equivalent detectable pressure(NEDP) value of 29Pa. This NEDP is much lower than the hundreds of Pascals achieved with existing optical resonant structures such as etalons, fiber gratings and dielectric multilayer interference filters available for acoustic measurement. The featured high sensitivity allowed the microring resonator to detect the weak photoacoustic signals from micro- or submicroscale objects. The inherent superbroad bandwidth of the optical microring resonator combined with an optically focused scanning beam provided POPAM with high resolution in the axial as well as both lateral directions while the axial resolution of conventional photoacoustic microscopy (PAM) suffers from the limited bandwidth of PZT detectors. Furthermore, the broadband microring resonator showed similar sensitivity to that of our most sensitive PZT detector. The current POPAM system provides a lateral resolution of 5 μm and an axial resolution of 8 μm, comparable to that achieved by optical microscopy while presenting the unique contrast of optical absorption and functional information complementing other optical modalities. The 3D structure of microvasculature, including capillary networks, and even individual red blood cells have been discerned successfully in the proof-of-concept experiments on mouse bladders ex vivo and mouse ears in vivo. The potential of approximately GHz bandwidth of the microring resonator also might allow much higher resolution than shown here in microscopy of optical absorption and acoustic propagation

  17. Single Cell Traction Microscopy within 3D Collagen Matrices

    NASA Astrophysics Data System (ADS)

    Wu, Mingming

    2014-03-01

    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, our current understanding of cell-ECM and cell-cell mechanical interactions is largely derived from 2D traction force microscopy, in which cells are cultured on a flat substrate. It is now clear that what we learn about cellular behavior on a 2D substrate does not always apply to cells embedded within a 3D biomatrix. 3D traction microscopy is emerging for mapping traction fields of single cells embedded in 3D gel, but current methods cannot account for the fibrous and nonlinear properties of collagen gel. In this talk, I will present a forward computation algorithm that we have developed for 3D cell traction measurements within collagen gels. The application of this technology to understanding cancer migration and invasion will be discussed. This work is supported by the National Center for Research Resources (5R21RR025801-03, NIH) and the National Institute of General Medical Sciences (8 R21 GM103388-03,NIH), and the Cornell Center on the Microenvironment & Metastasis.

  18. Exploring protein-DNA interactions in 3D using in situ construction, manipulation, and visualization of individual DNA-dumbbells with optical traps, microfluidics, and fluorescence microscopy

    PubMed Central

    Forget, Anthony L.; Dombrowski, Christopher C.; Amitani, Ichiro; Kowalczykowski, Stephen C.

    2015-01-01

    In this Protocol, we describe a procedure to generate ‘DNA-dumbbells’ — single molecules of DNA with a microscopic bead attached at each end — and techniques for manipulating individual DNA-dumbbells. We also detail the design and fabrication of a microfluidic device (flow cell) used in conjunction with dual optical trapping to manipulate DNA-dumbbells and to visualize individual protein–DNA complexes by single-molecule epifluorescence microscopy. Our design of the flow cell enables the rapid movement of trapped molecules between laminar flow channels and a flow-free ‘reservoir’. The reservoir provides the means to examine formation of DNA–protein complexes in solution in the absence of external flow forces, while still maintaining a predetermined end-to-end extension of the DNA. These features facilitate examination of the role of three-dimensional DNA conformation and dynamics in protein–DNA interactions. Preparation of flow cells and reagents requires two days each; in situ DNA-dumbbell assembly and imaging of single protein–DNA complexes requires another day. PMID:23411634

  19. Visualization of 3D optical lattices

    NASA Astrophysics Data System (ADS)

    Lee, Hoseong; Clemens, James

    2016-05-01

    We describe the visualization of 3D optical lattices based on Sisyphus cooling implemented with open source software. We plot the adiabatic light shift potentials found by diagonalizing the effective Hamiltonian for the light shift operator. Our program incorporates a variety of atomic ground state configurations with total angular momentum ranging from j = 1 / 2 to j = 4 and a variety of laser beam configurations including the two-beam lin ⊥ lin configuration, the four-beam umbrella configuration, and four beams propagating in two orthogonal planes. In addition to visualizing the lattice the program also evaluates lattice parameters such as the oscillation frequency for atoms trapped deep in the wells. The program is intended to help guide experimental implementations of optical lattices.

  20. Single molecule microscopy in 3D cell cultures and tissues.

    PubMed

    Lauer, Florian M; Kaemmerer, Elke; Meckel, Tobias

    2014-12-15

    From the onset of the first microscopic visualization of single fluorescent molecules in living cells at the beginning of this century, to the present, almost routine application of single molecule microscopy, the method has well-proven its ability to contribute unmatched detailed insight into the heterogeneous and dynamic molecular world life is composed of. Except for investigations on bacteria and yeast, almost the entire story of success is based on studies on adherent mammalian 2D cell cultures. However, despite this continuous progress, the technique was not able to keep pace with the move of the cell biology community to adapt 3D cell culture models for basic research, regenerative medicine, or drug development and screening. In this review, we will summarize the progress, which only recently allowed for the application of single molecule microscopy to 3D cell systems and give an overview of the technical advances that led to it. While initially posing a challenge, we finally conclude that relevant 3D cell models will become an integral part of the on-going success of single molecule microscopy. PMID:25453259

  1. 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.

  2. High Resolution, Large Deformation 3D Traction Force Microscopy

    PubMed Central

    López-Fagundo, Cristina; Reichner, Jonathan; Hoffman-Kim, Diane; Franck, Christian

    2014-01-01

    Traction Force Microscopy (TFM) is a powerful approach for quantifying cell-material interactions that over the last two decades has contributed significantly to our understanding of cellular mechanosensing and mechanotransduction. In addition, recent advances in three-dimensional (3D) imaging and traction force analysis (3D TFM) have highlighted the significance of the third dimension in influencing various cellular processes. Yet irrespective of dimensionality, almost all TFM approaches have relied on a linear elastic theory framework to calculate cell surface tractions. Here we present a new high resolution 3D TFM algorithm which utilizes a large deformation formulation to quantify cellular displacement fields with unprecedented resolution. The results feature some of the first experimental evidence that cells are indeed capable of exerting large material deformations, which require the formulation of a new theoretical TFM framework to accurately calculate the traction forces. Based on our previous 3D TFM technique, we reformulate our approach to accurately account for large material deformation and quantitatively contrast and compare both linear and large deformation frameworks as a function of the applied cell deformation. Particular attention is paid in estimating the accuracy penalty associated with utilizing a traditional linear elastic approach in the presence of large deformation gradients. PMID:24740435

  3. 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

  4. 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

  5. 3D resolution enhancement of deep-tissue imaging based on virtual spatial overlap modulation microscopy.

    PubMed

    Su, I-Cheng; Hsu, Kuo-Jen; Shen, Po-Ting; Lin, Yen-Yin; Chu, Shi-Wei

    2016-07-25

    During the last decades, several resolution enhancement methods for optical microscopy beyond diffraction limit have been developed. Nevertheless, those hardware-based techniques typically require strong illumination, and fail to improve resolution in deep tissue. Here we develop a high-speed computational approach, three-dimensional virtual spatial overlap modulation microscopy (3D-vSPOM), which immediately solves the strong-illumination issue. By amplifying only the spatial frequency component corresponding to the un-scattered point-spread-function at focus, plus 3D nonlinear value selection, 3D-vSPOM shows significant resolution enhancement in deep tissue. Since no iteration is required, 3D-vSPOM is much faster than iterative deconvolution. Compared to non-iterative deconvolution, 3D-vSPOM does not need a priori information of point-spread-function at deep tissue, and provides much better resolution enhancement plus greatly improved noise-immune response. This method is ready to be amalgamated with two-photon microscopy or other laser scanning microscopy to enhance deep-tissue resolution. PMID:27464077

  6. Quantifying cellular interaction dynamics in 3-D fluorescence microscopy data

    PubMed Central

    Klauschen, Frederick; Ishii, Masaru; Qi, Hai; Bajénoff, Marc; Egen, Jackson G.; Germain, Ronald N.; Meier-Schellersheim, Martin

    2012-01-01

    The wealth of information available from advanced fluorescence imaging techniques used to analyze biological processes with high spatial and temporal resolution calls for high-throughput image analysis methods. Here, we describe a fully automated approach to analyzing cellular interaction behavior in 3-D fluorescence microscopy images. As example application we present the analysis of drug-induced and S1P1-knock-out-related changes in bone-osteoclast interactions. Moreover, we apply our approach to images showing the spatial association of dendritic cells with the fibroblastic reticular cell network within lymph nodes and to microscopy data about T-B lymphocyte synapse formation. Such analyses that yield important information about the molecular mechanisms determining cellular interaction behavior would be very difficult to perform with approaches that rely on manual/semi-automated analyses. This protocol integrates adaptive threshold segmentation, object detection, adaptive color channel merging and neighborhood analysis and permits rapid, standardized, quantitative analysis and comparison of the relevant features in large data sets. PMID:19696749

  7. 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

  8. 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.

  9. 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.

  10. Atomic-Resolution 3D Electron Microscopy with Dynamic Diffraction

    SciTech Connect

    O'Keefe, Michael A.; Downing, Kenneth H.; Wenk, Hans-Rudolf; Meisheng, Hu

    2005-02-15

    Achievement of atomic-resolution electron-beam tomography will allow determination of the three-dimensional structure of nanoparticles (and other suitable specimens) at atomic resolution. Three-dimensional reconstructions will yield ''section'' images that resolve atoms overlapped in normal electron microscope images (projections), resolving lighter atoms such as oxygen in the presence of heavier atoms, and atoms that lie on non-lattice sites such as those in non-periodic defect structures. Lower-resolution electron microscope tomography has been used to produce reconstructed 3D images of nanoparticles [1] but extension to atomic resolution is considered not to be straightforward. Accurate three-dimensional reconstruction from two-dimensional projections generally requires that intensity in the series of 2-D images be a monotonic function of the specimen structure (often specimen density, but in our case atomic potential). This condition is not satisfied in electron microscopy when specimens with strong periodicity are tilted close to zone-axis orientation and produce ''anomalous'' image contrast because of strong dynamic diffraction components. Atomic-resolution reconstructions from tilt series containing zone-axis images (with their contrast enhanced by strong dynamical scattering) can be distorted when the stronger zone-axis images overwhelm images obtained in other ''random'' orientations in which atoms do not line up in neat columns. The first demonstrations of 3-D reconstruction to atomic resolution used five zone-axis images from test specimens of staurolite consisting of a mix of light and heavy atoms [2,3]. Initial resolution was to the 1.6{angstrom} Scherzer limit of a JEOL-ARM1000. Later experiments used focal-series reconstruction from 5 to 10 images to produce staurolite images from the ARM1000 with resolution extended beyond the Scherzer limit to 1.38{angstrom} [4,5]. To obtain a representation of the three-dimensional structure, images were obtained

  11. 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.

  12. Blind Depth-variant Deconvolution of 3D Data in Wide-field Fluorescence Microscopy.

    PubMed

    Kim, Boyoung; Naemura, Takeshi

    2015-01-01

    This paper proposes a new deconvolution method for 3D fluorescence wide-field microscopy. Most previous methods are insufficient in terms of restoring a 3D cell structure, since a point spread function (PSF) is simply assumed as depth-invariant, whereas a PSF of microscopy changes significantly along the optical axis. A few methods that consider a depth-variant PSF have been proposed; however, they are impractical, since they are non-blind approaches that use a known PSF in a pre-measuring condition, whereas an imaging condition of a target image is different from that of the pre-measuring. To solve these problems, this paper proposes a blind approach to estimate depth-variant specimen-dependent PSF and restore 3D cell structure. It is shown by experiments on that the proposed method outperforms the previous ones in terms of suppressing axial blur. The proposed method is composed of the following three steps: First, a non-parametric averaged PSF is estimated by the Richardson Lucy algorithm, whose initial parameter is given by the central depth prediction from intensity analysis. Second, the estimated PSF is fitted to Gibson's parametric PSF model via optimization, and depth-variant PSFs are generated. Third, a 3D cell structure is restored by using a depth-variant version of a generalized expectation-maximization. PMID:25950821

  13. 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

  14. 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

  15. Performance and sensitivity evaluation of 3D spot detection methods in confocal microscopy.

    PubMed

    Štěpka, Karel; Matula, Pavel; Matula, Petr; Wörz, Stefan; Rohr, Karl; Kozubek, Michal

    2015-08-01

    Reliable 3D detection of diffraction-limited spots in fluorescence microscopy images is an important task in subcellular observation. Generally, fluorescence microscopy images are heavily degraded by noise and non-specifically stained background, making reliable detection a challenging task. In this work, we have studied the performance and parameter sensitivity of eight recent methods for 3D spot detection. The study is based on both 3D synthetic image data and 3D real confocal microscopy images. The synthetic images were generated using a simulator modeling the complete imaging setup, including the optical path as well as the image acquisition process. We studied the detection performance and parameter sensitivity under different noise levels and under the influence of uneven background signal. To evaluate the parameter sensitivity, we propose a novel measure based on the gradient magnitude of the F1 score. We measured the success rate of the individual methods for different types of the image data and found that the type of image degradation is an important factor. Using the F1 score and the newly proposed sensitivity measure, we found that the parameter sensitivity is not necessarily proportional to the success rate of a method. This also provided an explanation why the best performing method for synthetic data was outperformed by other methods when applied to the real microscopy images. On the basis of the results obtained, we conclude with the recommendation of the HDome method for data with relatively low variations in quality, or the Sorokin method for image sets in which the quality varies more. We also provide alternative recommendations for high-quality images, and for situations in which detailed parameter tuning might be deemed expensive. PMID:26033916

  16. 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.

  17. Design and fabrication of a freeform prism array for 3D microscopy.

    PubMed

    Li, Lei; Yi, Allen Y

    2010-12-01

    Traditional microscopes have limitations in obtaining true 3D (three-dimensional) stereovision. Although some optical microscopes have been developed for 3D vision, many of them are complex, expensive, or limited to transparent samples. In this research, a freeform optical prism array was designed and fabricated to achieve 3D stereo imaging capability for microscope and machine vision applications. To form clear stereo images from multiple directions simultaneously, freeform optical surface design was applied to the prisms. In a ray tracing operation to determine the optical performance of the freeform prisms, Taylor series was used to calculate the surface shape. The virtual image spot diagrams were generated by using ray tracing methods for both the freeform prisms and the regular prisms. The results showed that all the light rays can be traced back to a single point for the freeform prism, and aberration was much smaller than that of the regular prism. The ray spots formed by the freeform prisms were adequate for image formation. Furthermore, the freeform prism array was fabricated by using a combined ultraprecision diamond turning and slow tool servo broaching process in a single, uninterrupted operation. The slow tool servo process ensured that the relative tolerance among prisms is guaranteed by the precision of the ultraprecision machine without the need for assembly. Finally 3D imaging tests were conducted to verify the freeform prism array's optical performance. The principle of the freeform prism array investigated in this research can be applied to microscopy, machine vision, robotic sensing, and many other areas. PMID:21119746

  18. 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. PMID:26073969

  19. An Optically Controlled 3D Cell Culturing System

    PubMed Central

    Ishii, Kelly S.; Hu, Wenqi; Namekar, Swapnil A.; Ohta, Aaron T.

    2012-01-01

    A novel 3D cell culture system was developed and tested. The cell culture device consists of a microfluidic chamber on an optically absorbing substrate. Cells are suspended in a thermoresponsive hydrogel solution, and optical patterns are utilized to heat the solution, producing localized hydrogel formation around cells of interest. The hydrogel traps only the desired cells in place while also serving as a biocompatible scaffold for supporting the cultivation of cells in 3D. This is demonstrated with the trapping of MDCK II and HeLa cells. The light intensity from the optically induced hydrogel formation does not significantly affect cell viability. PMID:22701475

  20. Multimodal interferometric microscopy for label-free 3D imaging of live cells in flow (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Shaked, Natan Tzvi

    2016-03-01

    I present multimodal wide-field interferometric microscopy platform for label-free 3-D imaging of live cells during fast flow. Using holographic optical tweezers, multiple cells can be optically trapped and rapidity rotated on all axes, while acquired using an external off-axis wide-field interferometric module developed in our lab. The interferometric projections are rapidly processed into the 3-D refractive-index profile of the cells using a tomographic phase microscopy algorithms that take into consideration optical diffraction effects. The algorithms for the 3-D refractive-index reconstruction, and for calculating various morphological parameters that should serve for online sorting of cells, are efficiently implemented in a nearly real-time manner. The potential of this new high-throughput imaging technique is for label-free image analysis and sorting of cells during flow, to substitute current cell sorting devices, which are based on external labeling that eventually damages the cell sample.

  1. 3D X-ray ultra-microscopy of bone tissue.

    PubMed

    Langer, M; Peyrin, F

    2016-02-01

    We review the current X-ray techniques with 3D imaging capability at the nano-scale: transmission X-ray microscopy, ptychography and in-line phase nano-tomography. We further review the different ultra-structural features that have so far been resolved: the lacuno-canalicular network, collagen orientation, nano-scale mineralization and their use as basis for mechanical simulations. X-ray computed tomography at the micro-metric scale is increasingly considered as the reference technique in imaging of bone micro-structure. The trend has been to push towards increasingly higher resolution. Due to the difficulty of realizing optics in the hard X-ray regime, the magnification has mainly been due to the use of visible light optics and indirect detection of the X-rays, which limits the attainable resolution with respect to the wavelength of the visible light used in detection. Recent developments in X-ray optics and instrumentation have allowed to implement several types of methods that achieve imaging that is limited in resolution by the X-ray wavelength, thus enabling computed tomography at the nano-scale. We review here the X-ray techniques with 3D imaging capability at the nano-scale: transmission X-ray microscopy, ptychography and in-line phase nano-tomography. Further, we review the different ultra-structural features that have so far been resolved and the applications that have been reported: imaging of the lacuno-canalicular network, direct analysis of collagen orientation, analysis of mineralization on the nano-scale and use of 3D images at the nano-scale to drive mechanical simulations. Finally, we discuss the issue of going beyond qualitative description to quantification of ultra-structural features. PMID:26370826

  2. 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

  3. 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.

  4. Tomographic phase microscopy using optical tweezers

    NASA Astrophysics Data System (ADS)

    Habaza, Mor; Gilboa, Barak; Roichman, Yael; Shaked, Natan T.

    2015-07-01

    We review our technique for tomographic phase microscopy with optical tweezers [1]. This tomographic phase microscopy approach enables full 3-D refractive-index reconstruction. Tomographic phase microscopy measures quantitatively the 3- D distribution of refractive-index in biological cells. We integrated our external interferometric module with holographic optical tweezers for obtaining quantitative phase maps of biological samples from a wide range of angles. The close-tocommon- path, off-axis interferometric system enables a full-rotation tomographic acquisition of a single cell using holographic optical tweezers for trapping and manipulating with a desired array of traps, while acquiring phase information of a single cell from all different angles and maintaining the native surrounding medium. We experimentally demonstrated two reconstruction algorithms: the filtered back-projection method and the Fourier diffraction method for 3-D refractive index imaging of yeast cells.

  5. 3D fluorescence anisotropy imaging using selective plane illumination microscopy.

    PubMed

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

    2015-08-24

    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

  6. 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

  7. Cortical Mapping of 3D Optical Topography in Infants

    PubMed Central

    Papademetriou, Maria D; Richards, John; Correia, Teresa; Blasi, Anna; Murphy, D. G.; Lloyd-Fox, Sarah; Johnson, Mark; Elwell, Clare E

    2014-01-01

    Precise localization of cortical activation in the early development of the infant brain remains unclear. It is challenging to co-register haemodynamic responses during functional activation in infants with the underlying anatomy of the brain. We used a multispectral imaging algorithm to reconstruct 3D optical topographic images of haemodynamic responses in an infant during voice processing. In this paper we present a method for co-registering 3D optical topography images reconstructed from functional activation data in infants onto anatomical brain images obtained from MRI structurals of the individual infants. PMID:23852529

  8. 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.

  9. 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.

  10. 3D optical tomography in the presence of void regions

    NASA Astrophysics Data System (ADS)

    Riley, J.; Dehghani, Hamid; Schweiger, Martin; Arridge, Simon R.; Ripoll, Jorge; Nieto-Vesperinas, Manuel

    2000-12-01

    We present an investigation of the effect of a 3D non-scattering gap region on image reconstruction in diffuse optical tomography. The void gap is modelled by the Radiosity-Diffusion method and the inverse problem is solved using the adjoint field method. The case of a sphere with concentric spherical gap is used as an example.

  11. 3D optical tomography in the presence of void regions.

    PubMed

    Riley, J; Dehghani, H; Schweiger, M; Arridge, S; Ripoll, J; Nieto-Vesperinas, M

    2000-12-18

    We present an investigation of the effect of a 3D non-scattering gap region on image reconstruction in diffuse optical tomography. The void gap is modelled by the Radiosity-Diffusion method and the inverse problem is solved using the adjoint field method. The case of a sphere with concentric spherical gap is used as an example. PMID:19407898

  12. 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

  13. 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

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. Optical Sensors and Methods for Underwater 3D Reconstruction.

    PubMed

    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

  19. 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

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. Resolution-limited optical recording in 3D.

    PubMed

    Orlic, Susanna; Dietz, Enrico; Frohmann, Sven; Rass, Jens

    2011-08-15

    We present an optical write/read system for high density optical data storage in 3-D. The microholographic approach relies on submicron-sized reflection gratings that encode the digital data. As in conventional optical data storage, the physical limitations are imposed by both the diffraction of light and resolution of the recording material. We demonstrate resolution-limited volume recording in photopolymer materials sensitive in the green and violet spectral range. The volume occupied by a micrograting scales down by the transition in the write/read wavelength. Readout yields a micrograting width of 306 nm at 532 nm and 197 nm at 405 nm. To our knowledge these are the smallest volume holograms ever recorded. The recordings demonstrate the potential of the technique for volumetric optical structuring, data storage and encryption. PMID:21934972

  5. Integrated optical 3D digital imaging based on DSP scheme

    NASA Astrophysics Data System (ADS)

    Wang, Xiaodong; Peng, Xiang; Gao, Bruce Z.

    2008-03-01

    We present a scheme of integrated optical 3-D digital imaging (IO3DI) based on digital signal processor (DSP), which can acquire range images independently without PC support. This scheme is based on a parallel hardware structure with aid of DSP and field programmable gate array (FPGA) to realize 3-D imaging. In this integrated scheme of 3-D imaging, the phase measurement profilometry is adopted. To realize the pipeline processing of the fringe projection, image acquisition and fringe pattern analysis, we present a multi-threads application program that is developed under the environment of DSP/BIOS RTOS (real-time operating system). Since RTOS provides a preemptive kernel and powerful configuration tool, with which we are able to achieve a real-time scheduling and synchronization. To accelerate automatic fringe analysis and phase unwrapping, we make use of the technique of software optimization. The proposed scheme can reach a performance of 39.5 f/s (frames per second), so it may well fit into real-time fringe-pattern analysis and can implement fast 3-D imaging. Experiment results are also presented to show the validity of proposed scheme.

  6. 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

  7. 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

  8. 3D Human cartilage surface characterization by optical coherence tomography.

    PubMed

    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 surface

  9. 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

  10. The impact of novel 3D diffraction optics development

    NASA Astrophysics Data System (ADS)

    Firsov, Alexander; Brzhezinskaya, Maria; Loechel, Heike; Siewert, Frank; Erko, Alexei

    2013-05-01

    Dedicated diffractive VUV- and X-ray optical elements are essential for future developments in synchrotron instrumentation and methods like e.g. time-resolved spectroscopy. The quality of optical components like gratings or diffractive focusing elements matters directly to the results achievable. On the other hand the availability of such optical components is very limited at present. In this contribution we report on the development of new methods of time-resolved x-ray spectroscopy based on novel 3D diffractive optical elements (DOE) with a unique combination of properties. Such optical elements are of highest interest for application in modern synchrotron facilities like Free Electron Lasers (FELs) as well as for laboratory facilities with high harmonic generators (HHG). The project includes theoretical work as well as the development of a dedicated technology, including metrology, to manufacture such type of optics for applications in atomic, molecular and condensed matter physics. The here discussed type of optics was successfully implemented for soft-X-ray-application at the femto-second-slicing beamline at BESSY II storage ring of the Helmholtz Zentrum Berlin. DOE are expected to be important components in beamlines at upcoming new high brilliance X-ray sources such as FELs. The application of DOE`s allows to reduce the number of optical elements in a beamline. Thus allow to provide the highest possible transmission and flux as well as preserving the unique properties of FEĹs, like wave-front and coherence.

  11. 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

  12. Multimodal 3D cancer-mimicking optical phantom.

    PubMed

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

    2016-02-01

    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

  13. 3D dosimetry by optical-CT scanning

    PubMed Central

    Oldham, Mark

    2007-01-01

    The need for an accurate, practical, low-cost 3D dosimetry system is becoming ever more critical as modern dose delivery techniques increase in complexity and sophistication. A recent report from the Radiological Physics Center (RPC) (1), revealed that 38% of institutions failed the head-and-neck IMRT phantom credentialing test at the first attempt. This was despite generous passing criteria (within 7% dose-difference or 4mm distance-to-agreement) evaluated at a half-dozen points and a single axial plane. The question that arises from this disturbing finding is – what percentage of institutions would have failed if a comprehensive 3D measurement had been feasible, rather than measurements restricted to the central film-plane and TLD points? This question can only be adequately answered by a comprehensive 3D-dosimetry system, which presents a compelling argument for its development as a clinically viable low cost dosimetry solution. Optical-CT dosimetry is perhaps the closest system to providing such a comprehensive solution. In this article, we review the origins and recent developments of optical-CT dosimetry systems. The principle focus is on first generation systems known to have highest accuracy but longer scan times. PMID:17460781

  14. 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

  15. Human tooth pulp anatomy visualization by 3D magnetic resonance microscopy

    PubMed Central

    Sustercic, Dusan; Sersa, Igor

    2012-01-01

    Background Precise assessment of dental pulp anatomy is of an extreme importance for a successful endodontic treatment. As standard radiographs of teeth provide very limited information on dental pulp anatomy, more capable methods are highly appreciated. One of these is 3D magnetic resonance (MR) microscopy of which diagnostic capabilities in terms of a better dental pulp anatomy assessment were evaluated in the study. Materials and methods Twenty extracted human teeth were scanned on a 2.35 T MRI system for MR microscopy using the 3D spin-echo method that enabled image acquisition with isotropic resolution of 100 μm. The 3D images were then post processed by ImageJ program (NIH) to obtain advanced volume rendered views of dental pulps. Results MR microscopy at 2.35 T provided accurate data on dental pulp anatomy in vitro. The data were presented as a sequence of thin 2D slices through the pulp in various orientations or as volume rendered 3D images reconstructed form arbitrary view-points. Sequential 2D images enabled only an approximate assessment of the pulp, while volume rendered 3D images were more precise in visualization of pulp anatomy and clearly showed pulp diverticles, number of pulp canals and root canal anastomosis. Conclusions This in vitro study demonstrated that MR microscopy could provide very accurate 3D visualization of dental pulp anatomy. A possible future application of the method in vivo may be of a great importance for the endodontic treatment. PMID:22933973

  16. 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.

  17. 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.

  18. 3D resolved mapping of optical aberrations in thick tissues

    PubMed Central

    Zeng, Jun; Mahou, Pierre; Schanne-Klein, Marie-Claire; Beaurepaire, Emmanuel; Débarre, Delphine

    2012-01-01

    We demonstrate a simple method for mapping optical aberrations with 3D resolution within thick samples. The method relies on the local measurement of the variation in image quality with externally applied aberrations. We discuss the accuracy of the method as a function of the signal strength and of the aberration amplitude and we derive the achievable resolution for the resulting measurements. We then report on measured 3D aberration maps in human skin biopsies and mouse brain slices. From these data, we analyse the consequences of tissue structure and refractive index distribution on aberrations and imaging depth in normal and cleared tissue samples. The aberration maps allow the estimation of the typical aplanetism region size over which aberrations can be uniformly corrected. This method and data pave the way towards efficient correction strategies for tissue imaging applications. PMID:22876353

  19. 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.

  20. 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.

  1. 2D and 3D X-Ray Structural Microscopy Using Submicron-Resolution Laue Microdiffraction

    SciTech Connect

    Budai, John D.; Yang, Wenge; Larson, Bennett C.; Tischler, Jonathan Z.; Liu, Wenjun; Ice, Gene E.

    2010-11-10

    We have developed a scanning, polychromatic x-ray microscopy technique with submicron spatial resolution at the Advanced Photon Source. In this technique, white undulator radiation is focused to submicron diameter using elliptical mirrors. Laue diffraction patterns scattered from the sample are collected with an area detector and then analyzed to obtain the local crystal structure, lattice orientation, and strain tensor. These new microdiffraction capabilities have enabled both 2D and 3D structural studies of materials on mesoscopic length-scales of tenths-to-hundreds of microns. For thin samples such as deposited films, 2D structural maps are obtained by step-scanning the area of interest. For example, 2D x-ray microscopy has been applied in studies of the epitaxial growth of oxide films. For bulk samples, a 3D differential-aperture x-ray microscopy technique has been developed that yields the full diffraction information from each submicron volume element. The capabilities of 3D x-ray microscopy are demonstrated here with measurements of grain orientations and grain boundary motion in polycrystalline aluminum during 3D thermal grain growth. X-ray microscopy provides the needed, direct link between the experimentally measured 3D microstructural evolution and the results of theory and modeling of materials processes on mesoscopic length scales.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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

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

    PubMed Central

    Mendez, Bernardino M.; Chiodo, Michael V.; Vandevender, Darl

    2016-01-01

    Summary: 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

  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. 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

  10. 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.

  11. 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.

  12. 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

  13. 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

  14. 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

  15. Parsing optical scanned 3D data by Bayesian inference

    NASA Astrophysics Data System (ADS)

    Xiong, Hanwei; Xu, Jun; Xu, Chenxi; Pan, Ming

    2015-10-01

    Optical devices are always used to digitize complex objects to get their shapes in form of point clouds. The results have no semantic meaning about the objects, and tedious process is indispensable to segment the scanned data to get meanings. The reason for a person to perceive an object correctly is the usage of knowledge, so Bayesian inference is used to the goal. A probabilistic And-Or-Graph is used as a unified framework of representation, learning, and recognition for a large number of object categories, and a probabilistic model defined on this And-Or-Graph is learned from a relatively small training set per category. Given a set of 3D scanned data, the Bayesian inference constructs a most probable interpretation of the object, and a semantic segment is obtained from the part decomposition. Some examples are given to explain the method.

  16. 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

  17. 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.

  18. Quantitative analysis of platelets aggregates in 3D by digital holographic microscopy

    PubMed Central

    Boudejltia, Karim Zouaoui; Ribeiro de Sousa, Daniel; Uzureau, Pierrick; Yourassowsky, Catherine; Perez-Morga, David; Courbebaisse, Guy; Chopard, Bastien; Dubois, Frank

    2015-01-01

    Platelet spreading and retraction play a pivotal role in the platelet plugging and the thrombus formation. In routine laboratory, platelet function tests include exhaustive information about the role of the different receptors present at the platelet surface without information on the 3D structure of platelet aggregates. In this work, we develop, a method in Digital Holographic Microscopy (DHM) to characterize the platelet and aggregate 3D shapes using the quantitative phase contrast imaging. This novel method is suited to the study of platelets physiology in clinical practice as well as the development of new drugs. PMID:26417523

  19. 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

  20. 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.

  1. 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

  2. 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

  3. 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.

  4. Full 3D morphology of diatoms flowing in a microfluidic channel by digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Savoia, Roberto; Memmolo, Pasquale; Merola, Francesco; Miccio, Lisa; D'Ippolito, Giuliana; Fontana, Angelo; Ferraro, Pietro

    2015-07-01

    In this paper, we present a new approach for three-dimensional reconstruction and biovolume estimation of some species of diatoms. An optofluidic platform, composed by an optical tweezer and holographic modulus, is employed to retrieve several holograms at different angular positions, which are processed by the shape from silhouette algorithm to estimate the 3D shape of the cells.

  5. An open-source deconvolution software package for 3-D quantitative fluorescence microscopy imaging

    PubMed Central

    SUN, Y.; DAVIS, P.; KOSMACEK, E. A.; IANZINI, F.; MACKEY, M. A.

    2010-01-01

    Summary Deconvolution techniques have been widely used for restoring the 3-D quantitative information of an unknown specimen observed using a wide-field fluorescence microscope. Deconv, an open-source deconvolution software package, was developed for 3-D quantitative fluorescence microscopy imaging and was released under the GNU Public License. Deconv provides numerical routines for simulation of a 3-D point spread function and deconvolution routines implemented three constrained iterative deconvolution algorithms: one based on a Poisson noise model and two others based on a Gaussian noise model. These algorithms are presented and evaluated using synthetic images and experimentally obtained microscope images, and the use of the library is explained. Deconv allows users to assess the utility of these deconvolution algorithms and to determine which are suited for a particular imaging application. The design of Deconv makes it easy for deconvolution capabilities to be incorporated into existing imaging applications. PMID:19941558

  6. Automated Identification and Localization of Hematopoietic Stem Cells in 3D Intravital Microscopy Data

    PubMed Central

    Khorshed, Reema A.; Hawkins, Edwin D.; Duarte, Delfim; Scott, Mark K.; Akinduro, Olufolake A.; Rashidi, Narges M.; Spitaler, Martin; Lo Celso, Cristina

    2015-01-01

    Summary Measuring three-dimensional (3D) localization of hematopoietic stem cells (HSCs) within the bone marrow microenvironment using intravital microscopy is a rapidly expanding research theme. This approach holds the key to understanding the detail of HSC-niche interactions, which are critical for appropriate stem cell function. Due to the complex tissue architecture of the bone marrow and to the progressive introduction of scattering and signal loss at increasing imaging depths, there is no ready-made software to handle efficient segmentation and unbiased analysis of the data. To address this, we developed an automated image analysis tool that simplifies and standardizes the biological interpretation of 3D HSC microenvironment images. The algorithm identifies HSCs and measures their localization relative to surrounding osteoblast cells and bone collagen. We demonstrate here the effectiveness, consistency, and accuracy of the proposed approach compared to current manual analysis and its wider applicability to analyze other 3D bone marrow components. PMID:26120058

  7. 3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy

    PubMed Central

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

    2015-01-01

    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 focused ion beam, 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 (TEM). Up to 1100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800–1500 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. PMID:24742339

  8. 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.

  9. 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

  10. 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.

  11. Spectral mapping of 3D multi-cellular tumor spheroids: time-resolved confocal microscopy.

    PubMed

    Mohapatra, Saswat; Nandi, Somen; Chowdhury, Rajdeep; Das, Gaurav; Ghosh, Surajit; Bhattacharyya, Kankan

    2016-07-21

    A tumor-like multi-cellular spheroid (3D) differs from a 2D cell in a number of ways. This is demonstrated using time resolved confocal microscopy. Two different tumor spheroids - HeLa (cervical cancer) and A549 (lung cancer) - are studied using 3 different fluorescent dyes - C153 (non-covalent), CPM (covalent) and doxorubicin (non-covalent, anti-cancer drug). The pattern of localization of these three fluorescent probes in the 3D tumor cell exhibits significant differences from that in the conventional 2D cells. For both the cells (HeLa and A549), the total uptake of doxorubicin in the 3D cell is much lower than that in the 2D cell. The uptake of doxorubicin molecules in the A549 spheroid is significantly different compared to the HeLa spheroid. The local polarity (i.e. emission maxima) and solvation dynamics in the 3D tumor cell differ from those in 2D cells. The covalent probe CPM exhibits intermittent fluorescence oscillations in the 1-2 s time scale. This is attributed to redox processes. These results may provide new insights into 3D tumors. PMID:27336201

  12. Dislocation Density Tensor Characterization of Deformation Using 3D X-Ray Microscopy

    SciTech Connect

    Larson, Ben C; Tischler, Jonathan Zachary; El-Azab, Anter; Liu, Wenjun

    2008-01-01

    Three-dimensional (3D) X-ray microscopy with submicron resolution has been used to make spatially resolved measurements of lattice curvature and elastic strain over two-dimensional slices in thin deformed Si plates. The techniques and capabilities associated with white-beam 3D X-ray microscopy are discussed, and both theoretical and experimental considerations associated with the measurement of Nye dislocation density tensors in deformed materials are presented. The ability to determine the local geometrically necessary dislocation (GND) density in the form of a dislocation density tensor, with micron spatial resolution over mesoscopic length scales, is demonstrated. Results are shown for the special case of an elastically bent (dislocation free) thin Si plate and for a similar thin Si plate that was bent plastically, above the brittle-to-ductile transition temperature, to introduce dislocations. Within the uncertainties of the measurements, the known result that GND density is zero for elastic bending is obtained, and well-defined GND distributions are observed in the plastically deformed Si plate. The direct and absolute connection between experimental measurements of GND density and multiscale modeling and computer simulations of deformation microstructures is discussed to highlight the importance of submicron-resolution 3D X-ray microscopy for mesoscale characterization of material defects and to achieve a fundamental understanding of deformation in ductile materials.

  13. Dislocation density tensor characterization of deformation using 3D x-ray microscopy.

    SciTech Connect

    Larson, B. C.; Tischler, J. Z.; El-Azab, A.; Liu, W.; ORNL; Florida State Univ.

    2008-04-01

    Three-dimensional (3D) X-ray microscopy with submicron resolution has been used to make spatially resolved measurements of lattice curvature and elastic strain over two-dimensional slices in thin deformed Si plates. The techniques and capabilities associated with white-beam 3D X-ray microscopy are discussed, and both theoretical and experimental considerations associated with the measurement of Nye dislocation density tensors in deformed materials are presented. The ability to determine the local geometrically necessary dislocation (GND) density in the form of a dislocation density tensor, with micron spatial resolution over mesoscopic length scales, is demonstrated. Results are shown for the special case of an elastically bent (dislocation free) thin Si plate and for a similar thin Si plate that was bent plastically, above the brittle-to-ductile transition temperature, to introduce dislocations. Within the uncertainties of the measurements, the known result that GND density is zero for elastic bending is obtained, and well-defined GND distributions are observed in the plastically deformed Si plate. The direct and absolute connection between experimental measurements of GND density and multiscale modeling and computer simulations of deformation microstructures is discussed to highlight the importance of submicron-resolution 3D X-ray microscopy for mesoscale characterization of material defects and to achieve a fundamental understanding of deformation in ductile materials.

  14. 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.

  15. Study on portable optical 3D coordinate measuring system

    NASA Astrophysics Data System (ADS)

    Ren, Tongqun; Zhu, Jigui; Guo, Yinbiao

    2009-05-01

    A portable optical 3D coordinate measuring system based on digital Close Range Photogrammetry (CRP) technology and binocular stereo vision theory is researched. Three ultra-red LED with high stability is set on a hand-hold target to provide measuring feature and establish target coordinate system. Ray intersection based field directional calibrating is done for the intersectant binocular measurement system composed of two cameras by a reference ruler. The hand-hold target controlled by Bluetooth wireless communication is free moved to implement contact measurement. The position of ceramic contact ball is pre-calibrated accurately. The coordinates of target feature points are obtained by binocular stereo vision model from the stereo images pair taken by cameras. Combining radius compensation for contact ball and residual error correction, object point can be resolved by transfer of axes using target coordinate system as intermediary. This system is suitable for on-field large-scale measurement because of its excellent portability, high precision, wide measuring volume, great adaptability and satisfying automatization. It is tested that the measuring precision is near to +/-0.1mm/m.

  16. Infrared differential interference contrast microscopy for overlay metrology on 3D-interconnect bonded wafers

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

    Overlay metrology for stacked layers will be playing a key role in bringing 3D IC devices into manufacturing. However, such bonded wafer pairs present a metrology challenge for optical microscopy tools by the opaque nature of silicon. Using infrared microscopy, silicon wafers become transparent to the near-infrared (NIR) wavelengths of the electromagnetic spectrum, enabling metrology at the interface of bonded wafer pairs. Wafers can be bonded face to face (F2F) or face to back (F2B) which the stacking direction is dictated by how the stacks are carried in the process and functionality required. For example, Memory stacks tend to use F2B stacking enables a better managed design. Current commercial tools use single image technique for F2F bonding overlay measurement because depth of focus is sufficient to include both surfaces; and use multiple image techniques for F2B overlay measurement application for the depth of focus is no longer sufficient to include both stacked wafer surfaces. There is a need to specify the Z coordinate or stacking wafer number through the silicon when visiting measurement wafer sites. Two shown images are of the same (X, Y) but separate Z location acquired at focus position of each wafer surface containing overlay marks. Usually the top surface image is bright and clear; however, the bottom surface image is somewhat darker and noisier as an adhesive layer is used in between to bond the silicon wafers. Thus the top and bottom surface images are further processed to achieve similar brightness and noise level before merged for overlay measurement. This paper presents a special overlay measurement technique, using the infrared differential interference contrast (DIC) microscopy technique to measure the F2B wafer bonding overlay by a single shot image. A pair of thinned wafers at 50 and 150 μm thickness is bonded on top of a carrier wafer to evaluate the bonding overlay. It works on the principle of interferometry to gain information about the

  17. A 3D Primary Vessel Reconstruction Framework with Serial Microscopy Images

    PubMed Central

    Liang, Yanhui; Wang, Fusheng; Treanor, Darren; Magee, Derek; Teodoro, George; Zhu, Yangyang; Kong, Jun

    2015-01-01

    Three dimensional microscopy images present significant potential to enhance biomedical studies. This paper presents an automated method for quantitative analysis of 3D primary vessel structures with histology whole slide images. With registered microscopy images of liver tissue, we identify primary vessels with an improved variational level set framework at each 2D slide. We propose a Vessel Directed Fitting Energy (VDFE) to provide prior information on vessel wall probability in an energy minimization paradigm. We find the optimal vessel cross-section associations along the image sequence with a two-stage procedure. Vessel mappings are first found between each pair of adjacent slides with a similarity function for four association cases. These bi-slide vessel components are further linked by Bayesian Maximum A Posteriori (MAP) estimation where the posterior probability is modeled as a Markov chain. The efficacy of the proposed method is demonstrated with 54 whole slide microscopy images of sequential sections from a human liver. PMID:26478919

  18. 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.

  19. 3D Axon structure extraction and analysis in confocal fluorescence microscopy images.

    PubMed

    Zhang, Yong; Zhou, Xiaobo; Lu, Ju; Lichtman, Jeff; Adjeroh, Donald; Wong, Stephen T C

    2008-08-01

    The morphological properties of axons, such as their branching patterns and oriented structures, are of great interest for biologists in the study of the synaptic connectivity of neurons. In these studies, researchers use triple immunofluorescent confocal microscopy to record morphological changes of neuronal processes. Three-dimensional (3D) microscopy image analysis is then required to extract morphological features of the neuronal structures. In this article, we propose a highly automated 3D centerline extraction tool to assist in this task. For this project, the most difficult part is that some axons are overlapping such that the boundaries distinguishing them are barely visible. Our approach combines a 3D dynamic programming (DP) technique and marker-controlled watershed algorithm to solve this problem. The approach consists of tracking and updating along the navigation directions of multiple axons simultaneously. The experimental results show that the proposed method can rapidly and accurately extract multiple axon centerlines and can handle complicated axon structures such as cross-over sections and overlapping objects. PMID:18336075

  20. 3D imaging by serial block face scanning electron microscopy for materials science using ultramicrotomy.

    PubMed

    Hashimoto, Teruo; Thompson, George E; Zhou, Xiaorong; Withers, Philip J

    2016-04-01

    Mechanical serial block face scanning electron microscopy (SBFSEM) has emerged as a means of obtaining three dimensional (3D) electron images over volumes much larger than possible by focused ion beam (FIB) serial sectioning and at higher spatial resolution than achievable with conventional X-ray computed tomography (CT). Such high resolution 3D electron images can be employed for precisely determining the shape, volume fraction, distribution and connectivity of important microstructural features. While soft (fixed or frozen) biological samples are particularly well suited for nanoscale sectioning using an ultramicrotome, the technique can also produce excellent 3D images at electron microscope resolution in a time and resource-efficient manner for engineering materials. Currently, a lack of appreciation of the capabilities of ultramicrotomy and the operational challenges associated with minimising artefacts for different materials is limiting its wider application to engineering materials. Consequently, this paper outlines the current state of the art for SBFSEM examining in detail how damage is introduced during slicing and highlighting strategies for minimising such damage. A particular focus of the study is the acquisition of 3D images for a variety of metallic and coated systems. PMID:26855205

  1. 3D positional tracking of ellipsoidal particles in a microtube flow using holographic microscopy

    NASA Astrophysics Data System (ADS)

    Byeon, Hyeok Jun; Seo, Kyung Won; Lee, Sang Joon

    2014-11-01

    Understanding of micro-scale flow phenomena is getting large attention under advances in micro-scale measurement technologies. Especially, the dynamics of particles suspended in a fluid is essential in both scientific and industrial fields. Moreover, most particles handled in research and industrial fields have non-spherical shapes rather than a simple spherical shape. Under various flow conditions, these non-spherical particles exhibit unique dynamic behaviors. To analyze these dynamic behaviors in a fluid flow, 3D positional information of the particles should be measured accurately. In this study, digital holographic microscopy (DHM) is employed to measure the 3D positional information of non-spherical particles, which are fabricated by stretching spherical polystyrene particles. 3D motions of those particles are obtained by interpreting the holograms captured from particles. Ellipsoidal particles with known size and shape are observed to verify the performance of the DHM technique. In addition, 3D positions of particles in a microtube flow are traced. This DHM technique exhibits promising potential in the analysis of dynamic behaviors of non-spherical particles suspended in micro-scale fluid flows.

  2. Computational methods for constructing protein structure models from 3D electron microscopy maps

    PubMed Central

    Esquivel-Rodríguez, Juan; Kihara, Daisuke

    2013-01-01

    Protein structure determination by cryo-electron microscopy (EM) has made significant progress in the past decades. Resolutions of EM maps have been improving as evidenced by recently reported structures that are solved at high resolutions close to 3 Å. Computational methods play a key role in interpreting EM data. Among many computational procedures applied to an EM map to obtain protein structure information, in this article we focus on reviewing computational methods that model protein three-dimensional (3D) structures from a 3D EM density map that is constructed from two-dimensional (2D) maps. The computational methods we discuss range from de novo methods, which identify structural elements in an EM map, to structure fitting methods, where known high resolution structures are fit into a low-resolution EM map. A list of available computational tools is also provided. PMID:23796504

  3. Steering knuckle diameter measurement based on optical 3D scanning

    NASA Astrophysics Data System (ADS)

    Song, Li-mei; Li, Da-peng; Chang, Yu-lan; Xi, Jiang-tao; Guo, Qing-hua

    2014-11-01

    To achieve accurate measurements, the creating a fitting hole for internal diameter (CFHID) measurement method and the establishing multi-sectional curve for external diameter (EMCED) measurement method are proposed in this paper, which are based on computer vision principle and three-dimensional (3D) reconstruction. The methods are able to highlight the 3D characteristics of the scanned object and to achieve the accurate measurement of 3D data. It can create favorable conditions for realizing the reverse design and 3D reconstruction of scanned object. These methods can also be applied to dangerous work environment or the occasion that traditional contact measurement can not meet the demands, and they can improve the security in measurement.

  4. 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

  5. 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

  6. Catching HIV ‘in the act’ with 3D electron microscopy

    PubMed Central

    Earl, Lesley A.; Lifson, Jeffrey D.; Subramaniam, Sriram

    2013-01-01

    The development of a safe, effective vaccine to prevent human immunodeficiency virus (HIV) infection is a key step for controlling the disease on a global scale. However, many aspects of HIV biology make vaccine design problematic, including the sequence diversity and structural variability of the surface envelope glycoproteins and the poor accessibility of neutralization-sensitive epitopes on the virus. In this review, we discuss recent progress in understanding HIV in a structural context using emerging tools in 3D electron microscopy, and outline how some of these advances could be important for a better understanding of mechanisms of viral entry and for vaccine design. PMID:23850373

  7. A new 3D tracking method for cell mechanics investigation exploiting the capabilities of digital holography in microscopy

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

    A method for 3D tracking has been developed exploiting Digital Holography features in Microscopy (DHM). 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 samples 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 amplitude refocusing in traditional tracking processes. Moreover, in biology and biomedical research fields one of the main topic is the understanding of morphology and mechanics of cells and microorganisms. Biological samples present low amplitude contrast that limits the information that can be retrieved through optical bright-field microscope measurements. The main effect on light propagating in such objects is in phase. This is known as phase-retardation or phase-shift. DHM is an innovative and alternative approach in microscopy, it's a good candidate for no-invasive and complete specimen analysis because its main characteristic is the possibility to discern between intensity and phase information performing quantitative mapping of the Optical Path Length. In this paper, the flexibility of DH is employed to analyze cell mechanics of unstained cells subjected to appropriate stimuli. DHM is used to measure all the parameters useful to understand the deformations induced by external and controlled stresses on in-vitro cells. Our configuration allows 3D tracking of micro-particles and, simultaneously, furnish quantitative phase-contrast maps. Experimental results are presented and discussed for in vitro cells.

  8. 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

  9. 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

  10. Optical 3D watermark based digital image watermarking for telemedicine

    NASA Astrophysics Data System (ADS)

    Li, Xiao Wei; Kim, Seok Tae

    2013-12-01

    Region of interest (ROI) of a medical image is an area including important diagnostic information and must be stored without any distortion. This algorithm for application of watermarking technique for non-ROI of the medical image preserving ROI. The paper presents a 3D watermark based medical image watermarking scheme. In this paper, a 3D watermark object is first decomposed into 2D elemental image array (EIA) by a lenslet array, and then the 2D elemental image array data is embedded into the host image. The watermark extraction process is an inverse process of embedding. The extracted EIA through the computational integral imaging reconstruction (CIIR) technique, the 3D watermark can be reconstructed. Because the EIA is composed of a number of elemental images possesses their own perspectives of a 3D watermark object. Even though the embedded watermark data badly damaged, the 3D virtual watermark can be successfully reconstructed. Furthermore, using CAT with various rule number parameters, it is possible to get many channels for embedding. So our method can recover the weak point having only one transform plane in traditional watermarking methods. The effectiveness of the proposed watermarking scheme is demonstrated with the aid of experimental results.

  11. Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision

    PubMed Central

    Kukulski, Wanda; Schorb, Martin; Welsch, Sonja; Picco, Andrea

    2011-01-01

    Correlative electron and fluorescence microscopy has the potential to elucidate the ultrastructural details of dynamic and rare cellular events, but has been limited by low precision and sensitivity. Here we present a method for direct mapping of signals originating from ∼20 fluorescent protein molecules to 3D electron tomograms with a precision of less than 100 nm. We demonstrate that this method can be used to identify individual HIV particles bound to mammalian cell surfaces. We also apply the method to image microtubule end structures bound to mal3p in fission yeast, and demonstrate that growing microtubule plus-ends are flared in vivo. We localize Rvs167 to endocytic sites in budding yeast, and show that scission takes place halfway through a 10-s time period during which amphiphysins are bound to the vesicle neck. This new technique opens the door for direct correlation of fluorescence and electron microscopy to visualize cellular processes at the ultrastructural scale. PMID:21200030

  12. Monitoring of Apoptosis in 3D Cell Cultures by FRET and Light Sheet Fluorescence Microscopy

    PubMed Central

    Weber, Petra; Schickinger, Sarah; Wagner, Michael; Angres, Brigitte; Bruns, Thomas; Schneckenburger, Herbert

    2015-01-01

    Non-radiative cell membrane associated Förster Resonance Energy Transfer (FRET) from an enhanced cyan fluorescent protein (ECFP) to an enhanced yellow fluorescent protein (EYFP) is used for detection of apoptosis in 3-dimensional cell cultures. FRET is visualized in multi-cellular tumor spheroids by light sheet based fluorescence microscopy in combination with microspectral analysis and fluorescence lifetime imaging (FLIM). Upon application of staurosporine and to some extent after treatment with phorbol-12-myristate-13-acetate (PMA), a specific activator of protein kinase c, the caspase-3 sensitive peptide linker DEVD is cleaved. This results in a reduction of acceptor (EYFP) fluorescence as well as a prolongation of the fluorescence lifetime of the donor (ECFP). Fluorescence spectra and lifetimes may, therefore, be used for monitoring of apoptosis in a realistic 3-dimensional system, while light sheet based microscopy appears appropriate for 3D imaging at low light exposure. PMID:25761242

  13. 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

  14. 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

  15. 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.

  16. 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.

  17. 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

  18. Four-directional stereo-microscopy for 3D particle tracking with real-time error evaluation.

    PubMed

    Hay, R F; Gibson, G M; Lee, M P; Padgett, M J; Phillips, D B

    2014-07-28

    High-speed video stereo-microscopy relies on illumination from two distinct angles to create two views of a sample from different directions. The 3D trajectory of a microscopic object can then be reconstructed using parallax to combine 2D measurements of its position in each image. In this work, we evaluate the accuracy of 3D particle tracking using this technique, by extending the number of views from two to four directions. This allows us to record two independent sets of measurements of the 3D coordinates of tracked objects, and comparison of these enables measurement and minimisation of the tracking error in all dimensions. We demonstrate the method by tracking the motion of an optically trapped microsphere of 5 μm in diameter, and find an accuracy of 2-5 nm laterally, and 5-10 nm axially, representing a relative error of less than 2.5% of its range of motion in each dimension. PMID:25089484

  19. Probing Local Mineralogy in 3D with Dual Energy X-Ray Microscopy

    NASA Astrophysics Data System (ADS)

    Gelb, J.; Yun, S.; Doerr, D.; Hunter, L.; Johnson, B.; Merkle, A.; Fahey, K.

    2013-12-01

    In recent years, 3D imaging of rock microstructures has become routine practice for determining pore-scale properties in the geosciences. X-Ray imaging techniques, such as X-Ray Microscopy (XRM), have demonstrated several unique capabilities: namely, the ability to characterize the same sample across a range of length scales and REVs (from millimeters to nanometers), and to perform these characterizations on the same sample over a range of times/treatments (e.g., to observe fluid transporting through the pore networks in a flow cell). While the XRM technique is a popular choice for structural (i.e., pore) characterization, historically it has provided little mineralogical information. This means that resulting simulations are either based on pore structure alone, or rely on correlative chemical mapping techniques for compositionally-sensitive models. Recent advancements in XRM techniques are now enabling compositional sensitivity for a variety of geological sample types. By collecting high-resolution 3D tomography data sets at two different source settings (energies), results may be mixed together to enhance the appearance (contrast) of specific materials. This approach is proving beneficial, for example, to mining applications to locate and identify precious metals, as well as for oil & gas applications to map local hydrophobicity. Here, we will introduce the technique of dual energy X-Ray microscopy, showing how it extends the capabilities of traditional XRM techniques, affording the same high resolution structural information while adding 3D compositional data. Application examples will be shown to illustrate its effectiveness at both the single to sub-micron length scale for mining applications as well as at the 150 nm length scale for shale rock analysis.

  20. 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

  1. Comparison of 3D orientation distribution functions measured with confocal microscopy and diffusion MRI.

    PubMed

    Schilling, Kurt; Janve, Vaibhav; Gao, Yurui; Stepniewska, Iwona; Landman, Bennett A; Anderson, Adam W

    2016-04-01

    The ability of diffusion MRI (dMRI) fiber tractography to non-invasively map three-dimensional (3D) anatomical networks in the human brain has made it a valuable tool in both clinical and research settings. However, there are many assumptions inherent to any tractography algorithm that can limit the accuracy of the reconstructed fiber tracts. Among them is the assumption that the diffusion-weighted images accurately reflect the underlying fiber orientation distribution (FOD) in the MRI voxel. Consequently, validating dMRI's ability to assess the underlying fiber orientation in each voxel is critical for its use as a biomedical tool. Here, using post-mortem histology and confocal microscopy, we present a method to perform histological validation of orientation functions in 3D, which has previously been limited to two-dimensional analysis of tissue sections. We demonstrate the ability to extract the 3D FOD from confocal z-stacks, and quantify the agreement between the MRI estimates of orientation information obtained using constrained spherical deconvolution (CSD) and the true geometry of the fibers. We find an orientation error of approximately 6° in voxels containing nearly parallel fibers, and 10-11° in crossing fiber regions, and note that CSD was unable to resolve fibers crossing at angles below 60° in our dataset. This is the first time that the 3D white matter orientation distribution is calculated from histology and compared to dMRI. Thus, this technique serves as a gold standard for dMRI validation studies - providing the ability to determine the extent to which the dMRI signal is consistent with the histological FOD, and to establish how well different dMRI models can predict the ground truth FOD. PMID:26804781

  2. Full-hemisphere automatic optical 3D measurement system

    NASA Astrophysics Data System (ADS)

    Kuehmstedt, Peter; Notni, Gunther; Schreiber, Wolfgang; Gerber, Joerg

    1997-09-01

    The measurement of 3D object shapes for the purpose of digitization of CAD-models and for the complete manufacturing control of components are important tasks of modern industrial inspection. The proposed 3D measurement system using structured-light illumination has the ability to avoid illumination-caused difficulties, like shadowing and excessive light intensities by light reflection and diffraction at the surface of the object, while measuring technical surfaces. For this purpose, the object under test is successively illuminated with a periodic grating structure from at least three different directions, using a telecentric projection system. At least three linearly independent phase-measurement values are measured by gray- code techniques to calculate the 3D coordinates of the object points. The experimental setup allows the determination of phase-measurement values with illuminations from up to 16 different directions. This is connected with a simultaneous variation of the intensity of the projected grating structures. Thus, areas of shadows are `shifted' across the object surface to spots where they have no influence on the result of the measurement, and also specular effects can be suppressed. Furthermore, in order to obtain the entire surface, the object to be digitized must be covered by many overlapping views taken from different directions. To view the entire surface, the object is moved into various measuring positions, using a second rotation axis. These views are merged within an object-centered coordinate system and are automatically rearranged into a uniform grid. For this purpose, a calibration procedure has been developed to measure absolute coordinates within a defined object coordinate system, so that the combination of the particular images is simple, because all measurements are performed within the same system of object coordinates. The power of this concept has been experimentally demonstrated, for example, by measuring the complete 3D shape

  3. 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.

  4. 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

  5. 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

  6. 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.

  7. 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

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

    PubMed

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

    2014-02-01

    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. PMID:24593372

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

    NASA Astrophysics Data System (ADS)

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

    2014-02-01

    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.

  10. 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.

  11. 3D optical Yagi–Uda nanoantenna array

    PubMed Central

    Dregely, Daniel; Taubert, Richard; Dorfmüller, Jens; Vogelgesang, Ralf; Kern, Klaus; Giessen, Harald

    2011-01-01

    Future photonic circuits with the capability of high-speed data processing at optical frequencies will rely on the implementation of efficient emitters and detectors on the nanoscale. Towards this goal, bridging the size mismatch between optical radiation and subwavelength emitters or detectors by optical nanoantennas is a subject of current research in the field of plasmonics. Here we introduce an array of three-dimensional optical Yagi–Uda antennas, fabricated using top-down fabrication techniques combined with layer-by-layer processing. We show that the concepts of radiofrequency antenna arrays can be applied to the optical regime proving superior directional properties compared with a single planar optical antenna, particularly for emission and reception into the third dimension. Measuring the optical properties of the structure reveals that impinging light on the array is efficiently absorbed on the subwavelength scale because of the high directivity. Moreover, we show in simulations that combining the array with suitable feeding circuits gives rise to the prospect of beam steering at optical wavelengths. PMID:21468019

  12. 3D optical Yagi-Uda nanoantenna array.

    PubMed

    Dregely, Daniel; Taubert, Richard; Dorfmüller, Jens; Vogelgesang, Ralf; Kern, Klaus; Giessen, Harald

    2011-01-01

    Future photonic circuits with the capability of high-speed data processing at optical frequencies will rely on the implementation of efficient emitters and detectors on the nanoscale. Towards this goal, bridging the size mismatch between optical radiation and subwavelength emitters or detectors by optical nanoantennas is a subject of current research in the field of plasmonics. Here we introduce an array of three-dimensional optical Yagi-Uda antennas, fabricated using top-down fabrication techniques combined with layer-by-layer processing. We show that the concepts of radiofrequency antenna arrays can be applied to the optical regime proving superior directional properties compared with a single planar optical antenna, particularly for emission and reception into the third dimension. Measuring the optical properties of the structure reveals that impinging light on the array is efficiently absorbed on the subwavelength scale because of the high directivity. Moreover, we show in simulations that combining the array with suitable feeding circuits gives rise to the prospect of beam steering at optical wavelengths. PMID:21468019

  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. Controlled 3D rotation of biological cells using optical multiple-force clamps

    PubMed Central

    Tanaka, Yoshio; Wakida, Shin-ich

    2014-01-01

    Controlled three-dimensional (3D) rotation of arbitrarily shaped objects in the observation space of optical microscopes is essential for realizing tomographic microscope imaging and offers great flexibility as a noncontact micromanipulation tool for biomedical applications. Herein, we present 3D rotational control of inhomogeneous biological samples using 3D optical multiple-force clamps based on time-shared scanning with a fast focus-tunable lens. For inhomogeneous samples with shape and optical anisotropy, we choose diatoms and their fragments, and demonstrate interactive and controlled 3D rotation about arbitrary axes in 3D Cartesian coordinates. We also outline the hardware setup and 3D rotation method for our demonstrations. PMID:25071968

  15. 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

  16. Multifocal multiphoton microscopy with adaptive optical correction

    NASA Astrophysics Data System (ADS)

    Coelho, Simao; Poland, Simon; Krstajic, Nikola; Li, David; Monypenny, James; Walker, Richard; Tyndall, David; Ng, Tony; Henderson, Robert; Ameer-Beg, Simon

    2013-02-01

    Fluorescence lifetime imaging microscopy (FLIM) is a well established approach for measuring dynamic signalling events inside living cells, including detection of protein-protein interactions. The improvement in optical penetration of infrared light compared with linear excitation due to Rayleigh scattering and low absorption have provided imaging depths of up to 1mm in brain tissue but significant image degradation occurs as samples distort (aberrate) the infrared excitation beam. Multiphoton time-correlated single photon counting (TCSPC) FLIM is a method for obtaining functional, high resolution images of biological structures. In order to achieve good statistical accuracy TCSPC typically requires long acquisition times. We report the development of a multifocal multiphoton microscope (MMM), titled MegaFLI. Beam parallelization performed via a 3D Gerchberg-Saxton (GS) algorithm using a Spatial Light Modulator (SLM), increases TCSPC count rate proportional to the number of beamlets produced. A weighted 3D GS algorithm is employed to improve homogeneity. An added benefit is the implementation of flexible and adaptive optical correction. Adaptive optics performed by means of Zernike polynomials are used to correct for system induced aberrations. Here we present results with significant improvement in throughput obtained using a novel complementary metal-oxide-semiconductor (CMOS) 1024 pixel single-photon avalanche diode (SPAD) array, opening the way to truly high-throughput FLIM.

  17. 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. PMID:26907410

  18. 3D imaging of mammalian cells with ion-abrasion scanning electron microscopy.

    PubMed

    Heymann, Jurgen A W; Shi, Dan; Kim, Sang; Bliss, Donald; Milne, Jacqueline L S; Subramaniam, Sriram

    2009-04-01

    Understanding the hierarchical organization of molecules and organelles within the interior of large eukaryotic cells is a challenge of fundamental interest in cell biology. We are using ion-abrasion scanning electron microscopy (IA-SEM) to visualize this hierarchical organization in an approach that combines focused ion-beam milling with scanning electron microscopy. Here, we extend our previous studies on imaging yeast cells to image subcellular architecture in human melanoma cells and melanocytes at resolutions as high as approximately 6 and approximately 20 nm in the directions parallel and perpendicular, respectively, to the direction of ion-beam milling. The 3D images demonstrate the striking spatial relationships between specific organelles such as mitochondria and membranes of the endoplasmic reticulum, and the distribution of unique cellular components such as melanosomes. We also show that 10nm-sized gold particles and quantum dot particles with 7 nm-sized cores can be detected in single cross-sectional images. IA-SEM is thus a useful tool for imaging large mammalian cells in their entirety at resolutions in the nanometer range. PMID:19116171

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

    PubMed

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

    2016-01-01

    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. PMID:27138688

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

    PubMed Central

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

    2016-01-01

    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. PMID:27138688

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

    PubMed

    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

  2. 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

  3. 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.

  4. 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.

  5. Deconvolution approach for 3D scanning microscopy with helical phase engineering.

    PubMed

    Roider, Clemens; Heintzmann, Rainer; Piestun, Rafael; Jesacher, Alexander

    2016-07-11

    RESCH (refocusing after scanning using helical phase engineering) microscopy is a scanning technique using engineered point spread functions which provides volumetric information. We present a strategy for processing the collected raw data with a multi-view maximum likelihood deconvolution algorithm, which inherently comprises the resolution gain of pixel-reassignment microscopy. The method, which we term MD-RESCH (for multi-view deconvolved RESCH), achieves in our current implementation a 20% resolution advantage along all three axes compared to RESCH and confocal microscopy. Along the axial direction, the resolution is comparable to that of image scanning microscopy. However, because the method inherently reconstructs a volume from a single 2D scan, a significantly higher optical sectioning becomes directly visible to the user, which would otherwise require collecting multiple 2D scans taken at a series of axial positions. Further, we introduce the use of a single-helical detection PSF to obtain an increased post-acquisition refocusing range. We present data from numerical simulations as well as experiments to confirm the validity of our approach. PMID:27410820

  6. 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

  7. 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

  8. 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.

  9. 3-D Adaptive Sparsity Based Image Compression With Applications to Optical Coherence Tomography.

    PubMed

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

    2015-06-01

    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

  10. 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

  11. Virtual 3D interactive system with embedded multiwavelength optical sensor array and sequential devices

    NASA Astrophysics Data System (ADS)

    Wang, Guo-Zhen; Huang, Yi-Pai; Hu, Kuo-Jui

    2012-06-01

    We proposed a virtual 3D-touch system by bare finger, which can detect the 3-axis (x, y, z) information of finger. This system has multi-wavelength optical sensor array embedded on the backplane of TFT panel and sequentail devices on the border of TFT panel. We had developed reflecting mode which can be worked by bare finger for the 3D interaction. A 4-inch mobile 3D-LCD with this proposed system was successfully been demonstrated already.

  12. Micro-optical system based 3D imaging for full HD depth image capturing

    NASA Astrophysics Data System (ADS)

    Park, Yong-Hwa; Cho, Yong-Chul; You, Jang-Woo; Park, Chang-Young; Yoon, Heesun; Lee, Sang-Hun; Kwon, Jong-Oh; Lee, Seung-Wan

    2012-03-01

    20 Mega-Hertz-switching high speed image shutter device for 3D image capturing and its application to system prototype are presented. For 3D image capturing, the system utilizes Time-of-Flight (TOF) principle by means of 20MHz high-speed micro-optical image modulator, so called 'optical shutter'. The high speed image modulation is obtained using the electro-optic operation of the multi-layer stacked structure having diffractive mirrors and optical resonance cavity which maximizes the magnitude of optical modulation. The optical shutter device is specially designed and fabricated realizing low resistance-capacitance cell structures having small RC-time constant. The optical shutter is positioned in front of a standard high resolution CMOS image sensor and modulates the IR image reflected from the object to capture a depth image. Suggested novel optical shutter device enables capturing of a full HD depth image with depth accuracy of mm-scale, which is the largest depth image resolution among the-state-of-the-arts, which have been limited up to VGA. The 3D camera prototype realizes color/depth concurrent sensing optical architecture to capture 14Mp color and full HD depth images, simultaneously. The resulting high definition color/depth image and its capturing device have crucial impact on 3D business eco-system in IT industry especially as 3D image sensing means in the fields of 3D camera, gesture recognition, user interface, and 3D display. This paper presents MEMS-based optical shutter design, fabrication, characterization, 3D camera system prototype and image test results.

  13. A 3D glass optrode array for optical neural stimulation

    PubMed Central

    Abaya, T.V.F.; Blair, S.; Tathireddy, P.; Rieth, L.; Solzbacher, F.

    2012-01-01

    This paper presents optical characterization of a first-generation SiO2 optrode array as a set of penetrating waveguides for both optogenetic and infrared (IR) neural stimulation. Fused silica and quartz discs of 3-mm thickness and 50-mm diameter were micromachined to yield 10 × 10 arrays of up to 2-mm long optrodes at a 400-μm pitch; array size, length and spacing may be varied along with the width and tip angle. Light delivery and loss mechanisms through these glass optrodes were characterized. Light in-coupling techniques include using optical fibers and collimated beams. Losses involve Fresnel reflection, coupling, scattering and total internal reflection in the tips. Transmission efficiency was constant in the visible and near-IR range, with the highest value measured as 71% using a 50-μm multi-mode in-coupling fiber butt-coupled to the backplane of the device. Transmittance and output beam profiles of optrodes with different geometries was investigated. Length and tip angle do not affect the amount of output power, but optrode width and tip angle influence the beam size and divergence independently. Finally, array insertion in tissue was performed to demonstrate its robustness for optical access in deep tissue. PMID:23243561

  14. Novel implementations of optical switch control module and 3D-CSP for 10 Gbps active optical access system

    NASA Astrophysics Data System (ADS)

    Wakayama, Koji; Okuno, Michitaka; Matsuoka, Yasunobu; Hosomi, Kazuhiko; Sagawa, Misuzu; Sugawara, Toshiki

    2009-11-01

    We propose an optical switch control procedure for high-performance and cost-effective 10 Gbps Active Optical Access System (AOAS) in which optical switches are used instead of optical splitters in PON (Passive Optical Network). We demonstrate the implemented optical switch control module on Optical Switching Unit (OSW) with logic circuits works effectively. We also propose a compact optical 3D-CSP (Chip Scale Package) to achieve the high performance of AOAS without losing cost advantage of PON. We demonstrate the implemented 3D-CSP works effectively.

  15. 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.

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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

  1. 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

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

    PubMed

    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 μm(3)) surrounding a large dendritic spine, a cylinder (~43 μm(3)) surrounding an oblique dendritic segment (3.4 μm long), and a parallelepiped (~178 μm(3)) 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

  3. 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

  4. 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

  5. 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.

  6. 3D bit-oriented optical storage in photopolymers

    NASA Astrophysics Data System (ADS)

    Orlic, Susanna; Ulm, Steffen; Eichler, Hans Joachim

    2001-01-01

    The bit-oriented data storage of conventional optical disks may be expanded into the third dimension by using microscopic reflection gratings instead of pits. Microgratings are holographically induced in a photopolymer layer. The Bragg selectivity of holographic volume gratings makes the application of multiplexing methods possible. High storage density may be achieved by combining wavelength multiplexing and multilayer storage. Wavelength multiplexing is realized by recording several gratings overlapping with write beams of different wavelengths. All gratings are recorded simultaneously in the same volume element. By translating a photopolymer sample perpendicularly to the laser beam axis during exposure, stripe-shaped microgratings are induced dynamically with constant linear velocity. The length of a grating is defined by the exposure time. Stripe-shaped gratings are required to realize an areal structure of recorded data similar to the pit-land structure of conventional disks that allows us to overtake the progress made in proceeding CD/DVD technology. Experimental results have been obtained for writing and reading of microholograms in different photopolymer materials including DuPont holographic recording films and CROP photopolymers from Polaroid.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 3D microscope imaging robust to restoration artifacts introduced by optically thick specimens

    NASA Astrophysics Data System (ADS)

    Patwary, Nurmohammed; King, Sharon V.; Preza, Chrysanthe

    2015-03-01

    We demonstrate 3D microscope imaging using computational optical sectioning microscopy (COSM) with an engineered point-spread function (PSF) robust to depth-induced spherical aberration (SA). Earlier we demonstrated that wavefront encoding (WFE) using a squared cubic (SQUBIC) phase mask reduces the PSF depth-variance in the presence of SA and that space-invariant (SI) restoration of simulated images using a single WFE-PSF does not lead to artifacts as in the conventional case. In this study, we show experimental verification of our WFE COSM approach. The WFE system used is a commercial microscope with a modified side port imaging path, where a spatial light modulator projects the SQUBIC phase mask on the back focal plane of the imaging lens. High resolution images of a test sample with 6 μm in diameter microspheres embedded in UV-cured optical cement (RI = 1.47) were captured using both the engineered and the conventional imaging paths of the system. The acquired images were restored using a regularized SI expectation maximization algorithm based on Tikhonov-Miller regularization with a roughness penalty. A comparative study quantified in terms of the correlation coefficients between the XZ medial sections of the restored images, from experimental data, shows an 11% reduction in depth sensitivity in the SQUBIC system compared to the conventional system.

  12. Fast 3D dark-field reflection-mode photoacoustic microscopy in vivo with a 30-MHz ultrasound linear array

    PubMed Central

    Song, Liang; Maslov, Konstantin; Bitton, Rachel; Shung, K. Kirk; Wang, Lihong V.

    2009-01-01

    We present an in vivo dark-field reflection-mode photoacoustic microscopy system that performs cross-sectional (B-scan) imaging at 50 Hz with realtime beamforming and 3D imaging consisting of 166 B-scan frames at 1 Hz with post-beamforming. To our knowledge, this speed is currently the fastest in photoacoustic imaging. A custom-designed light delivery system is integrated with a 30-MHz ultrasound linear array to realize dark-field reflection-mode imaging. Linear mechanical scanning of the array produces 3D images. The system has axial, lateral, and elevational resolutions of 25, 70, and 200 μm, respectively, and can image 3 mm deep in scattering biological tissues. Volumetric images of subcutaneous vasculature in rats are demonstrated in vivo. Fast 3D photoacoustic microscopy is anticipated to facilitate applications of photoacoustic imaging in biomedical studies that involve dynamics and clinical procedures that demand immediate diagnosis. PMID:19021408

  13. Developments in optical coherence microscopy

    NASA Astrophysics Data System (ADS)

    Rolland, J. P.; Meemon, P.; Thompson, K. P.; Murali, S.; Lee, K. S.

    2010-11-01

    Optical Coherence Microscopy (OCM) utilizes a high NA microscope objective in the sample arm to achieve an axially and laterally high resolution OCT image. An increase in NA, however, leads to a dramatically decreased depth of focus (DOF), and hence shortens the imaging depth range so that high lateral resolution is maintained only within a small depth region around the focal plane. One solution to increase the depth of imaging while keeping a high lateral resolution is dynamic-focusing. Utilizing the voltage controlled refocus capability of a liquid lens, we have recently presented a solution for invariant high resolution imaging using the liquid lens embedded within a fixed optics hand-held custom microscope designed specifically for optical imaging systems using a broadband light source centered at 800 nm with a 120 nm bandwidth. Subsequently, we have developed a Gabor-Domain Optical Coherence Microscopy (GD-OCM) that utilizes the high speed imaging of spectral domain OCT, the high lateral resolution of OCM, and the ability of real time refocusing of our custom design variable focus objective. Finally, key developments in Phase-Resolved Doppler OCT (PR-DOCT) are key enablers to combine high-resolution structural imaging with functional imaging. In this paper we review achievements in GD-OCM and detail how portions of in-focus cross-sectional images can be extracted and fused to form an invariant lateral resolution image with multiple cross-sectional images acquired corresponding to a discrete refocusing step along depth enabled by the varifocal device. We demonstrate sub-cellular resolution imaging of an African frog tadpole (Xenopus Laevis) taken from a 500 μm × 500 μm cross-section as well as cellular imaging in in vivo skin. Finally, A novel dual-detection full-range Fourier-domain optical coherence tomography system was developed that provides 7 μm axial resolution (in air) at about 90 kHz axial scan rate for mirror-image phase resolved Doppler imaging

  14. Next generation 3-D OFDM based optical access networks using FEC under various system impairments

    NASA Astrophysics Data System (ADS)

    Kumar, Pravindra; Srivastava, Anand

    2013-12-01

    Passive optical network based on orthogonal frequency division multiplexing (OFDM-PON) exhibits excellent performance in optical access networks due to its greater resistance to fiber dispersion, high spectral efficiency and exibility on both multiple services and dynamic bandwidth allocation. The major elements of conventional OFDM communication system are two-dimensional (2-D) signal mapper and one-dimensional (1-D) inverse fast fourier transform (IFFT). Three dimensional (3-D) OFDM use the concept of 3-D signal mapper and 2-D IFFT. With 3-D OFDM, minimum Euclidean distance (MED) is increased which results in BER performance improvement. As bit error rate (BER) depends on minimum Euclidean distance (MED) which is 15.46 % more in case of 3-D OFDM as compared to 2-D OFDM. Forward error correction (FEC) coding is a technique where redundancy is added to original bit sequence to increase the reliability of communication system. In this paper, we propose and analytically analyze a new PON architecture based on 3-D OFDM with convolutional coding and Viterbi decoding and is compared with conventional 2-D OFDM under various system impairments for coherent optical orthogonal frequency division multiplexing (CO-OFDM) without using any optical dispersion compensation. Analytical result show that at BER of 10-9, there is 2.7 dB, 3.8 dB and 9.3 dB signal-to-noise ratio (SNR) gain with 3-D OFDM, 3-D OFDM combined with convolutional coding and Viterbi hard decision decoding (CC-HDD) and 3-D OFDM combined with convolutional coding and Viterbi soft decision decoding (CC-SDD) respectively as compared to 2-D OFDM-PON. At BER of 10-9, 3-D OFDM-PON with CC-HDD gives 2.8 dB improvement in optical budget for both upstream and downstream path and gives 5.7 dB improvement in optical budget using 3-D OFDM-PON combined with CC-SDD as compared to conventional OFDM-PON system.

  15. General approach for the description of optical 3D measuring systems

    NASA Astrophysics Data System (ADS)

    Andrae, Peter; Jueptner, Werner P. O.; Kebbel, Volker; Osten, Wolfgang

    1997-07-01

    In this paper a general geometric description of the optical methods for 3D coordinate measurement is presented. Similar to holographic interferometry this new approach is based on the concept of measuring sensitivity. As a special case the derived basic relation is applied to the fringe projection technique using a physical model of this measurement method. Moreover a geometric 3D model that contributes to a dramatic reduction of systematic distortions of measured 3D coordinates is presented. On the one hand this model is sufficiently general but on the other hand still easy to handle. It permits an explicit and direct determination of 3D coordinates from primary measuring data as well as a calibration of the measuring set-up using linear identification methods mainly. The described 3D model can be applied also with advantage to multiview registration tasks.

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

    PubMed

    Krstajić, Nikola; Doran, Simon J

    2007-07-01

    3D measurement of optical attenuation is of interest in a variety of fields of biomedical importance, including spectrophotometry, optical projection tomography (OPT) and analysis of 3D radiation dosimeters. Accurate, precise and economical 3D measurements of optical density (OD) are a crucial step in enabling 3D radiation dosimeters to enter wider use in clinics. Polymer gels and Fricke gels, as well as dosimeters not based around gels, have been characterized for 3D dosimetry over the last two decades. A separate problem is the verification of the best readout method. A number of different imaging modalities (magnetic resonance imaging (MRI), optical CT, x-ray CT and ultrasound) have been suggested for the readout of information from 3D dosimeters. To date only MRI and laser-based optical CT have been characterized in detail. This paper describes some initial steps we have taken in establishing charge coupled device (CCD)-based optical CT as a viable alternative to MRI for readout of 3D radiation dosimeters. The main advantage of CCD-based optical CT over traditional laser-based optical CT is a speed increase of at least an order of magnitude, while the simplicity of its architecture would lend itself to cheaper implementation than both MRI and laser-based optical CT if the camera itself were inexpensive enough. Specifically, we study the following aspects of optical metrology, using high quality test targets: (i) calibration and quality of absorbance measurements and the camera requirements for 3D dosimetry; (ii) the modulation transfer function (MTF) of individual projections; (iii) signal-to-noise ratio (SNR) in the projection and reconstruction domains; (iv) distortion in the projection domain, depth-of-field (DOF) and telecentricity. The principal results for our current apparatus are as follows: (i) SNR of optical absorbance in projections is better than 120:1 for uniform phantoms in absorbance range 0.3 to 1.6 (and better than 200:1 for absorbances 1.0 to

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

    NASA Astrophysics Data System (ADS)

    Krstajic, Nikola; Doran, Simon J.

    2007-07-01

    3D measurement of optical attenuation is of interest in a variety of fields of biomedical importance, including spectrophotometry, optical projection tomography (OPT) and analysis of 3D radiation dosimeters. Accurate, precise and economical 3D measurements of optical density (OD) are a crucial step in enabling 3D radiation dosimeters to enter wider use in clinics. Polymer gels and Fricke gels, as well as dosimeters not based around gels, have been characterized for 3D dosimetry over the last two decades. A separate problem is the verification of the best readout method. A number of different imaging modalities (magnetic resonance imaging (MRI), optical CT, x-ray CT and ultrasound) have been suggested for the readout of information from 3D dosimeters. To date only MRI and laser-based optical CT have been characterized in detail. This paper describes some initial steps we have taken in establishing charge coupled device (CCD)-based optical CT as a viable alternative to MRI for readout of 3D radiation dosimeters. The main advantage of CCD-based optical CT over traditional laser-based optical CT is a speed increase of at least an order of magnitude, while the simplicity of its architecture would lend itself to cheaper implementation than both MRI and laser-based optical CT if the camera itself were inexpensive enough. Specifically, we study the following aspects of optical metrology, using high quality test targets: (i) calibration and quality of absorbance measurements and the camera requirements for 3D dosimetry; (ii) the modulation transfer function (MTF) of individual projections; (iii) signal-to-noise ratio (SNR) in the projection and reconstruction domains; (iv) distortion in the projection domain, depth-of-field (DOF) and telecentricity. The principal results for our current apparatus are as follows: (i) SNR of optical absorbance in projections is better than 120:1 for uniform phantoms in absorbance range 0.3 to 1.6 (and better than 200:1 for absorbances 1.0 to

  18. Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy.

    PubMed

    Höhn, K; Fuchs, J; Fröber, A; Kirmse, R; Glass, B; Anders-Össwein, M; Walther, P; Kräusslich, H-G; Dietrich, C

    2015-08-01

    In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV-pulsed mature human dendritic cells. PMID:25786567

  19. Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy

    PubMed Central

    HÖHN, K.; FUCHS, J.; FRÖBER, A.; KIRMSE, R.; GLASS, B.; ANDERS‐ÖSSWEIN, M.; WALTHER, P.; KRÄUSSLICH, H.‐G.

    2015-01-01

    Summary In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV‐pulsed mature human dendritic cells. PMID:25786567

  20. Automatic building detection and 3D shape recovery from single monocular electro-optic imagery

    NASA Astrophysics Data System (ADS)

    Lavigne, Daniel A.; Saeedi, Parvaneh; Dlugan, Andrew; Goldstein, Norman; Zwick, Harold

    2007-04-01

    The extraction of 3D building geometric information from high-resolution electro-optical imagery is becoming a key element in numerous geospatial applications. Indeed, producing 3D urban models is a requirement for a variety of applications such as spatial analysis of urban design, military simulation, and site monitoring of a particular geographic location. However, almost all operational approaches developed over the years for 3D building reconstruction are semiautomated ones, where a skilled human operator is involved in the 3D geometry modeling of building instances, which results in a time-consuming process. Furthermore, such approaches usually require stereo image pairs, image sequences, or laser scanning of a specific geographic location to extract the 3D models from the imagery. Finally, with current techniques, the 3D geometric modeling phase may be characterized by the extraction of 3D building models with a low accuracy level. This paper describes the Automatic Building Detection (ABD) system and embedded algorithms currently under development. The ABD system provides a framework for the automatic detection of buildings and the recovery of 3D geometric models from single monocular electro-optic imagery. The system is designed in order to cope with multi-sensor imaging of arbitrary viewpoint variations, clutter, and occlusion. Preliminary results on monocular airborne and spaceborne images are provided. Accuracy assessment of detected buildings and extracted 3D building models from single airborne and spaceborne monocular imagery of real scenes are also addressed. Embedded algorithms are evaluated for their robustness to deal with relatively dense and complicated urban environments.

  1. 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

  2. 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.

  3. 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.

  4. Focused ion beam 3D nano-patterned optical fiber tips for advanced beam profile engineering

    NASA Astrophysics Data System (ADS)

    Janeiro, Ricardo; Flores, Raquel; Ribeiro, Ana R.; Jorge, Pedro; Viegas, Jaime

    2015-03-01

    Focused ion beam (FIB) patterning of 3D topography on optical fiber tips for application in stand-alone, rugged and simplified setups for optical tweezers cell sorters, optical near-field lithography and optical beam profile engineering are reported. We demonstrate various configurations based on single-step FIB patterning, multiple-step FIB processing and hybrid approaches based on optical fiber pre- and post-FIB treatment with either etching, fusion splicing, photopolymerization or electroplating steps for optical fiber texture, topography and composition engineering. Different conductive coatings for minimal charge accumulation and beam drift are studied with the relative merits compared. Furthermore optimal beam parameters for accurate pattern replication and positioning are also presented. Measured experimental field profiles are compared with numerical simulations of fabricated optical fiber tips for fabrication accuracy evaluation. Applications employing these engineered fiber tips in the field of optical tweezers, optical vortex generation, photolithography, photo-polymerization and beam forming are presented.

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. Scanning Tunneling Optical Resonance Microscopy

    NASA Technical Reports Server (NTRS)

    Bailey, Sheila; Wilt, Dave; Raffaelle, Ryne; Gennett, Tom; Tin, Padetha; Lau, Janice; Castro, Stephanie; Jenkins, Philip; Scheiman, Dave

    2003-01-01

    Scanning tunneling optical resonance microscopy (STORM) is a method, now undergoing development, for measuring optoelectronic properties of materials and devices on the nanoscale by means of a combination of (1) traditional scanning tunneling microscopy (STM) with (2) tunable laser spectroscopy. In STORM, an STM tip probing a semiconductor is illuminated with modulated light at a wavelength in the visible-to-near-infrared range and the resulting photoenhancement of the tunneling current is measured as a function of the illuminating wavelength. The photoenhancement of tunneling current occurs when the laser photon energy is sufficient to excite charge carriers into the conduction band of the semiconductor. Figure 1 schematically depicts a proposed STORM apparatus. The light for illuminating the semiconductor specimen at the STM would be generated by a ring laser that would be tunable across the wavelength range of interest. The laser beam would be chopped by an achromatic liquid-crystal modulator. A polarization-maintaining optical fiber would couple the light to the tip/sample junction of a commercial STM. An STM can be operated in one of two modes: constant height or constant current. A STORM apparatus would be operated in the constant-current mode, in which the height of the tip relative to the specimen would be varied in order to keep the tunneling current constant. In this mode, a feedback control circuit adjusts the voltage applied to a piezoelectric actuator in the STM that adjusts the height of the STM tip to keep the tunneling current constant. The exponential relationship between the tunneling current and tip-to-sample distance makes it relatively easy to implement this mode of operation. The choice of method by which the photoenhanced portion of the tunneling current would be measured depends on choice of the frequency at which the input illumination would be modulated (chopped). If the frequency of modulation were low enough (typically < 10 Hz) that the

  10. Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

    PubMed Central

    Kim, Doory; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Zhuang, Xiaowei

    2015-01-01

    Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets. PMID:25874453

  11. 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

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

    PubMed

    Molaei, Mehdi; Sheng, Jian

    2014-12-29

    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

  13. 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

  14. 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

  15. Fast high-resolution 3D total internal reflection fluorescence microscopy by incidence angle scanning and azimuthal averaging

    PubMed Central

    Boulanger, Jérôme; Gueudry, Charles; Münch, Daniel; Cinquin, Bertrand; Paul-Gilloteaux, Perrine; Bardin, Sabine; Guérin, Christophe; Senger, Fabrice; Blanchoin, Laurent; Salamero, Jean

    2014-01-01

    Total internal reflection fluorescence microscopy (TIRFM) is the method of choice to visualize a variety of cellular processes in particular events localized near the plasma membrane of live adherent cells. This imaging technique not relying on particular fluorescent probes provides a high sectioning capability. It is, however, restricted to a single plane. We present here a method based on a versatile design enabling fast multiwavelength azimuthal averaging and incidence angles scanning to computationally reconstruct 3D images sequences. We achieve unprecedented 50-nm axial resolution over a range of 800 nm above the coverslip. We apply this imaging modality to obtain structural and dynamical information about 3D actin architectures. We also temporally decipher distinct Rab11a-dependent exocytosis events in 3D at a rate of seven stacks per second. PMID:25404337

  16. Ultra-high voltage electron microscopy of primitive algae illuminates 3D ultrastructures of the first photosynthetic eukaryote

    PubMed Central

    Takahashi, Toshiyuki; Nishida, Tomoki; Saito, Chieko; Yasuda, Hidehiro; Nozaki, Hisayoshi

    2015-01-01

    A heterotrophic organism 1–2 billion years ago enslaved a cyanobacterium to become the first photosynthetic eukaryote, and has diverged globally. The primary phototrophs, glaucophytes, are thought to retain ancestral features of the first photosynthetic eukaryote, but examining the protoplast ultrastructure has previously been problematic in the coccoid glaucophyte Glaucocystis due to its thick cell wall. Here, we examined the three-dimensional (3D) ultrastructure in two divergent species of Glaucocystis using ultra-high voltage electron microscopy. Three-dimensional modelling of Glaucocystis cells using electron tomography clearly showed that numerous, leaflet-like flattened vesicles are distributed throughout the protoplast periphery just underneath a single-layered plasma membrane. This 3D feature is essentially identical to that of another glaucophyte genus Cyanophora, as well as the secondary phototrophs in Alveolata. Thus, the common ancestor of glaucophytes and/or the first photosynthetic eukaryote may have shown similar 3D structures. PMID:26439276

  17. 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.

  18. Potential and limitations of microscopy and Raman spectroscopy for live-cell analysis of 3D cell cultures.

    PubMed

    Charwat, Verena; Schütze, Karin; Holnthoner, Wolfgang; Lavrentieva, Antonina; Gangnus, Rainer; Hofbauer, Pablo; Hoffmann, Claudia; Angres, Brigitte; Kasper, Cornelia

    2015-07-10

    Today highly complex 3D cell culture formats that closely mimic the in vivo situation are increasingly available. Despite their wide use, the development of analytical methods and tools that can work within the depth of 3D-tissue constructs lags behind. In order to get the most information from a 3D cell sample, adequate and reliable assays are required. However, the majority of tools and methods used today have been originally designed for 2D cell cultures and translation to a 3D environment is in general not trivial. Ideally, an analytical method should be non-invasive and allow for repeated observation of living cells in order to detect dynamic changes in individual cells within the 3D cell culture. Although well-established laser confocal microscopy can be used for these purposes, this technique has serious limitations including penetration depth and availability. Focusing on two relevant analytical methods for live-cell monitoring, we discuss the current challenges of analyzing living 3D samples: microscopy, which is the most widely used technology to observe and examine cell cultures, has been successfully adapted for 3D samples by recording of so-called "z-stacks". However the required equipment is generally very expensive and therefore access is often limited. Consequently alternative and less advanced approaches are often applied that cannot capture the full structural complexity of a 3D sample. Similarly, image analysis tools for quantification of microscopic images range from highly specialized and costly to simplified and inexpensive. Depending on the actual sample composition and scientific question the best approach needs to be assessed individually. Another more recently introduced technology for non-invasive cell analysis is Raman micro-spectroscopy. It enables label-free identification of cellular metabolic changes with high sensitivity and has already been successful applied to 2D and 3D cell cultures. However, its future significance for cell

  19. 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

  20. 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. PMID:24113529

  1. Atomic force microscopy imaging and 3-D reconstructions of serial thin sections of a single cell and its interior structures

    PubMed Central

    Chen, Yong; Cai, Jiye; Zhao, Tao; Wang, Chenxi; Dong, Shuo; Luo, Shuqian; Chen, Zheng W.

    2010-01-01

    The thin sectioning has been widely applied in electron microscopy (EM), and successfully used for an in situ observation of inner ultrastructure of cells. This powerful technique has recently been extended to the research field of atomic force microscopy (AFM). However, there have been no reports describing AFM imaging of serial thin sections and three-dimensional (3-D) reconstruction of cells and their inner structures. In the present study, we used AFM to scan serial thin sections approximately 60nm thick of a mouse embryonic stem (ES) cell, and to observe the in situ inner ultrastructure including cell membrane, cytoplasm, mitochondria, nucleus membrane, and linear chromatin. The high-magnification AFM imaging of single mitochondria clearly demonstrated the outer membrane, inner boundary membrane and cristal membrane of mitochondria in the cellular compartment. Importantly, AFM imaging on six serial thin sections of a single mouse ES cell showed that mitochondria underwent sequential changes in the number, morphology and distribution. These nanoscale images allowed us to perform 3-D surface reconstruction of interested interior structures in cells. Based on the serial in situ images, 3-D models of morphological characteristics, numbers and distributions of interior structures of the single ES cells were validated and reconstructed. Our results suggest that the combined AFM and serial-thin-section technique is useful for the nanoscale imaging and 3-D reconstruction of single cells and their inner structures. This technique may facilitate studies of proliferating and differentiating stages of stem cells or somatic cells at a nanoscale. PMID:15850704

  2. 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

  3. 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

  4. Disposable optics for microscopy diagnostics.

    PubMed

    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

  5. 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.

  6. 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

  7. The design of 3D optical system for multidirectional phase tomography

    NASA Astrophysics Data System (ADS)

    Antoš, Martin

    2008-12-01

    The design of 3D optical system for multidirectional phase tomograph is presented in detail. The suggested tomograph uses a multidirectional holographic interferometer with diffusive light. The method of dividing of the laser-beam to object and reference beams is described. The optimisation of geometrical dimensions of the testing area and optical parameters of projection beams was done in order to increase the number of obtainable angular projections. Finally, projecting properties of the scanning system of the tomograph are presented.

  8. 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

  9. Parameter optimization for through-focus scanning optical microscopy.

    PubMed

    Attota, Ravi Kiran; Kang, Hyeonggon

    2016-06-27

    It is important to economically and non-destructively analyze three-dimensional (3-D) shapes of nanometer to micrometer scale objects with sub-nanometer measurement resolution for emerging high-volume nanomanufacturing, especially for process control. High-throughput through-focus scanning optical microscopy (TSOM) demonstrates promise for such applications. TSOM uses a conventional optical microscope for 3-D shape metrology by making use of the complete set of through-focus, four-dimensional optical data. However, a systematic study showing the effect of various parameters on the TSOM method is lacking. Here we present the optimization of the basic parameters such as illumination numerical aperture (NA), collection NA, focus step height, digital camera pixel size, illumination polarization, and illumination wavelength to achieve peak performance of the TSOM method. PMID:27410642

  10. 3D single-molecule tracking using one- and two-photon excitation microscopy

    NASA Astrophysics Data System (ADS)

    Liu, Cong; Perillo, Evan P.; Zhuang, Quincy; Huynh, Khang T.; Dunn, Andrew K.; Yeh, Hsin-Chih

    2014-03-01

    Three dimensional single-molecule tracking (3D-SMT) has revolutionized the way we study fundamental cellular processes. By analyzing the spatial trajectories of individual molecules (e.g. a receptor or a signaling molecule) in 3D space, one can discern the internalization or transport dynamics of these molecules, study the heterogeneity of subcellular structures, and elucidate the complex spatiotemporal regulation mechanisms. Sub-diffraction localization precision, sub-millisecond temporal resolution and tens-of-seconds observation period are the benchmarks of current 3D-SMT techniques. We have recently built two molecular tracking systems in our labs. The first system is a previously reported confocal tracking system, which we denote as the 1P-1E-4D (one-photon excitation, one excitation beam, and four fiber-coupled detectors) system. The second system is a whole new design that is based on two-photon excitation, which we denote as the 2P-4E-1D (two-photon excitation, four excitation beams, and only one detector) system. Here we compare these two systems based on Monte Carlo simulation of tracking a diffusing fluorescent molecule. Through our simulation, we have characterized the limitation of individual systems and optimized the system parameters such as magnification, z-plane separation, and feedback gains.

  11. Analysis of optical characteristics of photopolymer-based VHOE for multiview autostereoscopic 3D display system

    NASA Astrophysics Data System (ADS)

    Cho, Byung-Chul; Gu, Jung-Sik; Kim, Eun-Soo

    2002-06-01

    Generally, an autostereoscopic display presents a 3D image to a viewer without the need for glasses or other encumbering viewing aids. In this paper, we propose a new autostereoscopic 3D video display system which allows viewers to observe 3D images in the same range of viewing angle. In this system, a photopolymer-based VHOE is made from volume holographic recording materials and it is used for projecting a multiview images to the spatially different directions sequentially in time. Since this technique is based on the VHOE made from the photorefractive photopolymer instead of the conventional parallax barrier or lenticular sheet, the resolution and parallax number of the proposed VHOE-based 3D display system are limited by the photopolymer's physical and optical properties. To make the photopolymer to be applicable for a multiview autostereoscopic 3D display system, the photopolymer must be capable of achieving some properties such as a low distortion of the diffracted light beam, high diffraction efficiency, and uniform intensities of the reconstructed diffracted lights from the fully recorded diffraction gratings. In this paper, the optical and physical characteristics of the DuPont HRF photopolymer-based VHOE such as a distortion of displayed image, uniformity of the diffracted light intensity, photosensitivity and diffraction efficiency are measured and discussed.

  12. A physical model eye with 3D resolution test targets for optical coherence tomography

    NASA Astrophysics Data System (ADS)

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

    2014-09-01

    Optical coherence tomography (OCT) has been widely employed as non-invasive 3D imaging diagnostic instrument, particularly in the field of ophthalmology. Although OCT has been approved for use in clinic in USA, Europe and Asia, international standardization of this technology is still in progress. Validation of OCT imaging capabilities is considered extremely important to ensure its effective use in clinical diagnoses. Phantom with appropriate test targets can assist evaluate and calibrate imaging performance of OCT at both installation and throughout lifetime of the instrument. In this paper, we design and fabricate a physical model eye with 3D resolution test targets to characterize OCT imaging performance. The model eye was fabricated with transparent resin to simulate realistic ophthalmic testing environment, and most key optical elements including cornea, lens and vitreous body were realized. The test targets which mimic USAF 1951 test chart were fabricated on the fundus of the model eye by 3D printing technology. Differing from traditional two dimensional USAF 1951 test chart, a group of patterns which have different thickness in depth were fabricated. By measuring the 3D test targets, axial resolution as well as lateral resolution of an OCT system can be evaluated at the same time with this model eye. To investigate this specialized model eye, it was measured by a scientific spectral domain OCT instrument and a clinical OCT system respectively. The results demonstrate that the model eye with 3D resolution test targets have the potential of qualitatively and quantitatively validating the performance of OCT systems.

  13. 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

  14. 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.

  15. 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.

  16. Quantification of fluorescent spots in time series of 3D confocal microscopy images of endoplasmic reticulum exit sites based on the HMAX transform

    NASA Astrophysics Data System (ADS)

    Matula, Petr; Verissimo, Fatima; Wörz, Stefan; Eils, Roland; Pepperkok, Rainer; Rohr, Karl

    2010-03-01

    We present an approach for the quantification of fluorescent spots in time series of 3-D confocal microscopy images of endoplasmic reticulum exit sites of dividing cells. Fluorescent spots are detected based on extracted image regions of highest response using the HMAX transform and prior convolution of the 3-D images with a Gaussian kernel. The sensitivity of the involved parameters was studied and a quantitative evaluation using both 3-D synthetic and 3-D real data was performed. The approach was successfully applied to more than one thousand 3-D confocal microscopy images.

  17. A 3D visualization and guidance system for handheld optical imaging devices

    NASA Astrophysics Data System (ADS)

    Azar, Fred S.; de Roquemaurel, Benoit; Cerussi, Albert; Hajjioui, Nassim; Li, Ang; Tromberg, Bruce J.; Sauer, Frank

    2007-03-01

    We have developed a novel 3D visualization and guidance system for handheld optical imaging devices. In this paper, the system is applied to measurements of breast/cancerous tissue optical properties using a handheld diffuse optical spectroscopy (DOS) instrument. The combined guidance system/DOS instrument becomes particularly useful for monitoring neoadjuvant chemotherapy in breast cancer patients and for longitudinal studies where measurement reproducibility is critical. The system uses relatively inexpensive hardware components and comprises a 6 degrees-of-freedom (DOF) magnetic tracking device including a DC field generator, three sensors, and a PCI card running on a PC workstation. A custom-built virtual environment combined with a well-defined workflow provide the means for image-guided measurements, improved longitudinal studies of breast optical properties, 3D reconstruction of optical properties within the anatomical map, and serial data registration. The DOS instrument characterizes tissue function such as water, lipid and total hemoglobin concentration. The patient lies on her back at a 45-degrees angle. Each spectral measurement requires consistent contact with the skin, and lasts about 5-10 seconds. Therefore a limited number of positions may be studied. In a reference measurement session, the physician acquires surface points on the breast. A Delaunay-based triangulation algorithm is used to build the virtual breast surface from the acquired points. 3D locations of all DOS measurements are recorded. All subsequently acquired surfaces are automatically registered to the reference surface, thus allowing measurement reproducibility through image guidance using the reference measurements.

  18. 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

  19. 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

  20. Virtual touch 3D interactive system for autostereoscopic display with embedded optical sensor

    NASA Astrophysics Data System (ADS)

    Huang, Yi-Pai; Wang, Guo-Zhen; Ma, Ming-Ching; Tung, Shang-Yu; Huang, Shu-Yi; Tseng, Hung-Wei; Kuo, Chung-Hong; Li, Chun-Huai

    2011-06-01

    The traidational 3D interactive sysetm which uses CCD camera to capture image is difficult to operate on near range for mobile applications.Therefore, 3D interactive display with embedded optical sensor was proposed. Based on optical sensor based system, we proposed four different methods to support differenct functions. T mark algorithm can obtain 5- axis information (x, y, z,θ, and φ)of LED no matter where LED was vertical or inclined to panel and whatever it rotated. Sequential mark algorithm and color filter based algorithm can support mulit-user. Finally, bare finger touch system with sequential illuminator can achieve to interact with auto-stereoscopic images by bare finger. Furthermore, the proposed methods were verified on a 4-inch panel with embedded optical sensors.

  1. 3D Radiative Aspects of the Increased Aerosol Optical Depth Near Clouds

    NASA Technical Reports Server (NTRS)

    Marshak, Alexander; Wen, Guoyong; Remer, Lorraine; Cahalan, Robert; Coakley, Jim

    2007-01-01

    To characterize aerosol-cloud interactions it is important to correctly retrieve aerosol optical depth in the vicinity of clouds. It is well reported in the literature that aerosol optical depth increases with cloud cover. Part of the increase comes from real physics as humidification; another part, however, comes from 3D cloud effects in the remote sensing retrievals. In many cases it is hard to say whether the retrieved increased values of aerosol optical depth are remote sensing artifacts or real. In the presentation, we will discuss how the 3D cloud affects can be mitigated. We will demonstrate a simple model that can assess the enhanced illumination of cloud-free columns in the vicinity of clouds. This model is based on the assumption that the enhancement in the cloud-free column radiance comes from the enhanced Rayleigh scattering due to presence of surrounding clouds. A stochastic cloud model of broken cloudiness is used to simulate the upward flux.

  2. 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

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

    NASA Astrophysics Data System (ADS)

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

    2015-05-01

    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.

  4. 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. PMID:26769364

  5. 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

  6. Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy

    PubMed Central

    Park, HyunJoo; Hong, Sung-Hee; Kim, Kyoohyun; Cho, Shin-Hyeong; Lee, Won-Ja; Kim, Youngchan; Lee, Sang-Eun; Park, YongKeun

    2015-01-01

    Babesia microti causes “emergency” human babesiosis. However, little is known about the alterations in B. microti invaded red blood cells (Bm-RBCs) at the individual cell level. Through quantitative phase imaging techniques based on laser interferometry, we present the simultaneous measurements of structural, chemical, and mechanical modifications in individual mouse Bm-RBCs. 3-D refractive index maps of individual RBCs and in situ parasite vacuoles are imaged, from which total contents and concentration of dry mass are also precisely quantified. In addition, we examine the dynamic membrane fluctuation of Bm-RBCs, which provide information on cell membrane deformability. PMID:26039793

  7. 3D laser scanning microscopy of hypervelocity impact features in metal and aerogel targets

    NASA Astrophysics Data System (ADS)

    Hillier, J. K.; Postberg, F.; Price, M. C.; Trieloff, M.; Li, Y. W.; Srama, R.

    2012-09-01

    We present the results of a study into the mapping of hypervelocity impact features using a Keyence VK-X200 3D laser scanning microscope. The impact features observed are impact craters in a variety of different metal targets (Al, Au and Cu) and impact tracks in aerogel targets, similar to those used in the Stardust mission. Differences in crater morphology between different target materials and impact velocities, as well as differences in track depth and diameter in aerogel, for particles of known constant dimensions, are discussed.

  8. 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

  9. 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

  10. 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).

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

    PubMed

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

    2015-10-30

    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). PMID:26436905

  12. 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

  13. 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

  14. 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

  15. 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...

  16. 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...

  17. 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. PMID:26316044

  18. 3D imaging and quantitative analysis of small solubilized membrane proteins and their complexes by transmission electron microscopy

    PubMed Central

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

    2013-01-01

    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. PMID:23267047

  19. 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.

  20. 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

  1. 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. PMID:22051087

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

    PubMed

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

    2016-05-01

    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

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. Dynamic 3D imaging based on acousto-optic heterodyne fringe interferometry.

    PubMed

    Guan, Yingjian; Yin, Yongkai; Li, Ameng; Liu, Xiaoli; Peng, Xiang

    2014-06-15

    An acoustic-optics heterodyne fringe interferometry coupled with a three-camera system is developed for dynamic 3D imaging. In this system, first-order beams with a slight frequency difference diffracted from two acousto-optic deflectors (AODs) form a beat intensity fringe pattern. Setting the frequency of the trigger signal for the CCD cameras into four times the beat frequency, four-step phase-shifting fringe patterns can be obtained, and the wrapped phase map (WPM) can be calculated. Under the epipolar constraint among three cameras, the homologous points can be determined unambiguously with the assistant of a WPM; thus the 3D shape can be reconstructed while skipping the phase unwrapping step. Experimental results are presented to validate this approach. PMID:24978566

  9. Enhanced Optical Transmission Mediated by Localized Plasmons in Anisotropic, 3D Nanohole Arrays

    PubMed Central

    Yang, Jiun-Chan; Gao, Hanwei; Suh, Jae Yong; Zhou, Wei; Lee, Min Hyung; Odom, Teri W.

    2010-01-01

    This paper describes 3D nanohole arrays whose high optical transmission is mediated more by localized surface plasmon (LSP) excitations than by surface plasmon polaritons (SPPs). First, LSPs on 3D hole arrays lead to optical transmission an order of magnitude higher than 2D planar hole arrays. Second, LSP-mediated transmission is broadband and more tunable than SPP-enhanced transmission which is restricted by Bragg coupling. Third, for the first time, two types of surface plasmons can be selectively excited and manipulated on the same plasmonic substrate. This new plasmonic substrate fabricated by high-throughput nanolithography techniques paves the way for cutting-edge optoelectronic and biomedical applications. PMID:20698633

  10. 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

  11. 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.

  12. Increasing the filling of ultracold KRb molecules in a 3D optical lattice

    NASA Astrophysics Data System (ADS)

    Moses, Steven; Covey, Jacob; Gadway, Bryce; Yan, Bo; Miecnikowski, Matthew; Ye, Jun; Jin, Deborah

    2015-05-01

    Ultracold polar molecules, with their long-range electric dipolar interactions, offer new opportunities for studying quantum magnetism and many-body physics. Recently, our group observed spin exchange interactions between KRb molecules in a 3D optical lattice, which is one of the first steps towards studying lattice spin models with polar molecules. The lattice fillings were about 10% or less in these experiments. Future experiments will benefit greatly from lower entropies and higher lattice fillings. Here, we have investigated the molecular creation process in a 3D optical lattice with the goal of maximizing the filling fraction. We start by loading a BEC of Rb and a degenerate Fermi gas of K into a 3D optical lattice. In the absence of K, Rb is a Mott insulator. We study how the Mott insulator and the filling of Rb are affected by the presence of K and develop a strategy to maintain high Rb filling throughout the molecule production process. We also find that we can convert a large fraction of these Rb to molecules when we operate with low Rb numbers. We acknowledge funding from DARPA, DOE, NIST, NSF, AFOSR, and the NDSEG Graduate Fellowship.

  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 PAGESBeta

    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. 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

  16. 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

  17. 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.

  18. Dynamic Analysis of 2D Electromagnetic Resonant Optical Scanner Using 3D Finite Element Method

    NASA Astrophysics Data System (ADS)

    Hirata, Katsuhiro; Hong, Sara; Maeda, Kengo

    The optical scanner is a scanning device in which a laser beam is reflected by a mirror that can be rotated or oscillated. In this paper, we propose a new 2D electromagnetic resonant optical scanner that employs electromagnets and leaf springs. Torque characteristics and resonance characteristics of the scanner are analyzed using the 3D finite element method. The validity of the analysis is shown by comparing the characteristics inferred from the analysis with the characteristics of the prototype. Further, 2D resonance is investigated by introducing a superimposed-frequency current in a single coil.

  19. 3D optical metamaterials formed by holographic assembly and directed solidification of eutectics (Presentation Recording)

    NASA Astrophysics Data System (ADS)

    Braun, Paul V.

    2015-09-01

    Nanoscale integration of materials in three dimensions is critical for the realization of a number of highly functional optical metamaterials. Starting with structures enabled via eutectic solidification and holographic lithography, our team is applying unique template-based and post-synthetic materials transformations in conjunction with powerful computational design tools to develop the scientific underpinnings of, and to produce, 3D metamaterials derived from directionally solidified eutectics. Our approach involves close interactions among computational design, photonic theory, eutectic materials development, template fabrication, materials chemistry, and optical characterization.

  20. Study of hybrid driven micromirrors for 3-D variable optical attenuator applications.

    PubMed

    Koh, Kah How; Soon, Bo Woon; Tsai, Julius Minglin; Danner, Aaron J; Lee, Chengkuo

    2012-09-10

    Aluminium-coated micromirrors driven by electrothermal and electromagnetic actuations have been demonstrated for 3-D variable optical attenuation applications. Three types of attenuation schemes based on electrothermal, electromagnetic and hybrid, i.e. combination of electrothermal and electromagnetic, actuations have been developed. In addition, two different designs have been fabricated and characterized to investigate the effects of the variations made to both the actuators on the optical attenuation performances of the micromirror. Our unique design of using both ET and EM actuators simultaneously to achieve attenuation is the first demonstration of such hybrid driven CMOS compatible MEMS VOA device. PMID:23037278

  1. 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.

  2. 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.

  3. 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.

  4. First steps toward 3D high resolution imaging using adaptive optics and full-field optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Blanco, Leonardo; Blavier, Marie; Glanc, Marie; Pouplard, Florence; Tick, Sarah; Maksimovic, Ivan; Chenegros, Guillaume; Mugnier, Laurent; Lacombe, Francois; Rousset, Gérard; Paques, Michel; Le Gargasson, Jean-François; Sahel, Jose-Alain

    2008-09-01

    We describe here two parts of our future 3D fundus camera coupling Adaptive Optics and full-field Optical Coherence Tomography. The first part is an Adaptive Optics flood imager installed at the Quinze-Vingts Hospital, regularly used on healthy and pathological eyes. A posteriori image reconstruction is performed, increasing the final image quality and field of view. The instrument lateral resolution is better than 2 microns. The second part is a full-field Optical Coherence Tomograph, which has demonstrated capability of performing a simple kind of "4 phases" image reconstruction of non biological samples and ex situ retinas. Final aim is to couple both parts in order to achieve 3D high resolution mapping of in vivo retinas.

  5. CMOS array of photodiodes with electronic processing for 3D optical reconstruction

    NASA Astrophysics Data System (ADS)

    Hornero, Gemma; Montane, Enric; Chapinal, Genis; Moreno, Mauricio; Herms, Atila

    2001-04-01

    It is well known that laser time-of-flight (TOF) and optical triangulation are the most useful optical techniques for distance measurements. The first one is more suitable for large distances, since for short range of distances high modulation frequencies of laser diodes (©200-500MHz) are needed. For these ranges, optical triangulation is simpler, as it is only necessary to read the projection of the laser point over a linear optical sensor without any laser modulation. Laser triangulation is based on the rotation of the object. This motion shifts the projected point over the linear sensor, resulting on 3D information, by means of the whole readout of the linear sensor in each angle position. On the other hand, a hybrid method of triangulation and TOF can be implemented. In this case, a synchronized scanning of a laser beam over the object results in different arrival times of light to each pixel. The 3D information is carried by these delays. Only a single readout of the linear sensor is needed. In this work we present the design of two different linear arrays of photodiodes in CMOS technology, the first one based on the Optical triangulation measurement and the second one based in this hybrid method (TFO). In contrast to PSD (Position Sensitive Device) and CCDs, CMOS technology can include, on the same chip, photodiodes, control and processing electronics, that in the other cases should be implemented with external microcontrollers.

  6. 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.

  7. A joint estimation detection of Glaucoma progression in 3D spectral domain optical coherence tomography optic nerve head images

    PubMed Central

    Belghith, Akram; Bowd, Christopher; Weinreb, Robert N.; Zangwill, Linda M.

    2014-01-01

    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. PMID:25606299

  8. 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.

  9. Parallel deconvolution of large 3D images obtained by confocal laser scanning microscopy.

    PubMed

    Pawliczek, Piotr; Romanowska-Pawliczek, Anna; Soltys, Zbigniew

    2010-03-01

    Various deconvolution algorithms are often used for restoration of digital images. Image deconvolution is especially needed for the correction of three-dimensional images obtained by confocal laser scanning microscopy. Such images suffer from distortions, particularly in the Z dimension. As a result, reliable automatic segmentation of these images may be difficult or even impossible. Effective deconvolution algorithms are memory-intensive and time-consuming. In this work, we propose a parallel version of the well-known Richardson-Lucy deconvolution algorithm developed for a system with distributed memory and implemented with the use of Message Passing Interface (MPI). It enables significantly more rapid deconvolution of two-dimensional and three-dimensional images by efficiently splitting the computation across multiple computers. The implementation of this algorithm can be used on professional clusters provided by computing centers as well as on simple networks of ordinary PC machines. PMID:19725070

  10. 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.

  11. 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

  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 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

  13. Multiview 3-D Echocardiography Fusion with Breath-Hold Position Tracking Using an Optical Tracking System.

    PubMed

    Punithakumar, Kumaradevan; Hareendranathan, Abhilash R; McNulty, Alexander; Biamonte, Marina; He, Allen; Noga, Michelle; Boulanger, Pierre; Becher, Harald

    2016-08-01

    Recent advances in echocardiography allow real-time 3-D dynamic image acquisition of the heart. However, one of the major limitations of 3-D echocardiography is the limited field of view, which results in an acquisition insufficient to cover the whole geometry of the heart. This study proposes the novel approach of fusing multiple 3-D echocardiography images using an optical tracking system that incorporates breath-hold position tracking to infer that the heart remains at the same position during different acquisitions. In six healthy male volunteers, 18 pairs of apical/parasternal 3-D ultrasound data sets were acquired during a single breath-hold as well as in subsequent breath-holds. The proposed method yielded a field of view improvement of 35.4 ± 12.5%. To improve the quality of the fused image, a wavelet-based fusion algorithm was developed that computes pixelwise likelihood values for overlapping voxels from multiple image views. The proposed wavelet-based fusion approach yielded significant improvement in contrast (66.46 ± 21.68%), contrast-to-noise ratio (49.92 ± 28.71%), signal-to-noise ratio (57.59 ± 47.85%) and feature count (13.06 ± 7.44%) in comparison to individual views. PMID:27166019

  14. 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

  15. 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

  16. 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

  17. Seeing a Mycobacterium-Infected Cell in Nanoscale 3D: Correlative Imaging by Light Microscopy and FIB/SEM Tomography.

    PubMed

    Beckwith, Marianne Sandvold; Beckwith, Kai Sandvold; Sikorski, Pawel; Skogaker, Nan Tostrup; Flo, Trude Helen; Halaas, Øyvind

    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

  18. Application of the split-gradient method to 3D image deconvolution in fluorescence microscopy.

    PubMed

    Vicidomini, G; Boccacci, P; Diaspro, A; Bertero, M

    2009-04-01

    The methods of image deconvolution are important for improving the quality of the detected images in the different modalities of fluorescence microscopy such as wide-field, confocal, two-photon excitation and 4Pi. Because deconvolution is an ill-posed problem, it is, in general, reformulated in a statistical framework such as maximum likelihood or Bayes and reduced to the minimization of a suitable functional, more precisely, to a constrained minimization, because non-negativity of the solution is an important requirement. Next, iterative methods are designed for approximating such a solution. In this paper, we consider the Bayesian approach based on the assumption that the noise is dominated by photon counting, so the likelihood is of the Poisson-type, and that the prior is edge-preserving, as derived from a simple Markov random field model. By considering the negative logarithm of the a posteriori probability distribution, the computation of the maximum a posteriori (MAP) estimate is reduced to the constrained minimization of a functional that is the sum of the Csiszár I-divergence and a regularization term. For the solution of this problem, we propose an iterative algorithm derived from a general approach known as split-gradient method (SGM) and based on a suitable decomposition of the gradient of the functional into a negative and positive part. The result is a simple modification of the standard Richardson-Lucy algorithm, very easily implementable and assuring automatically the non-negativity of the iterates. Next, we apply this method to the particular case of confocal microscopy for investigating the effect of several edge-preserving priors proposed in the literature using both synthetic and real confocal images. The quality of the restoration is estimated both by computation of the Kullback-Leibler divergence of the restored image from the detected one and by visual inspection. It is observed that the noise artefacts are considerably reduced and desired

  19. Fast error simulation of optical 3D measurements at translucent objects

    NASA Astrophysics Data System (ADS)

    Lutzke, P.; Kühmstedt, P.; Notni, G.

    2012-09-01

    The scan results of optical 3D measurements at translucent objects deviate from the real objects surface. This error is caused by the fact that light is scattered in the objects volume and is not exclusively reflected at its surface. A few approaches were made to separate the surface reflected light from the volume scattered. For smooth objects the surface reflected light is dominantly concentrated in specular direction and could only be observed from a point in this direction. Thus the separation either leads to measurement results only creating data for near specular directions or provides data from not well separated areas. To ensure the flexibility and precision of optical 3D measurement systems for translucent materials it is necessary to enhance the understanding of the error forming process. For this purpose a technique for simulating the 3D measurement at translucent objects is presented. A simple error model is shortly outlined and extended to an efficient simulation environment based upon ordinary raytracing methods. In comparison the results of a Monte-Carlo simulation are presented. Only a few material and object parameters are needed for the raytracing simulation approach. The attempt of in-system collection of these material and object specific parameters is illustrated. The main concept of developing an error-compensation method based on the simulation environment and the collected parameters is described. The complete procedure is using both, the surface reflected and the volume scattered light for further processing.

  20. 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.

  1. Intraretinal Layer Segmentation of Macular Optical Coherence Tomography Images Using Optimal 3-D Graph Search

    PubMed Central

    Abràmoff, Michael D.; Kardon, Randy; Russell, Stephen R.; Wu, Xiaodong; Sonka, Milan

    2008-01-01

    Current techniques for segmenting macular optical coherence tomography (OCT) images have been 2-D in nature. Furthermore, commercially available OCT systems have only focused on segmenting a single layer of the retina, even though each intraretinal layer may be affected differently by disease. We report an automated approach for segmenting (anisotropic) 3-D macular OCT scans into five layers. Each macular OCT dataset consisted of six linear radial scans centered at the fovea. The six surfaces defining the five layers were identified on each 3-D composite image by transforming the segmentation task into that of finding a minimum-cost closed set in a geometric graph constructed from edge/regional information and a priori determined surface smoothness and interaction constraints. The method was applied to the macular OCT scans of 12 patients (24 3-D composite image datasets) with unilateral anterior ischemic optic neuropathy (AION). Using the average of three experts’ tracings as a reference standard resulted in an overall mean unsigned border positioning error of 6.1 ± 2.9 µm, a result comparable to the interobserver variability (6.9 ± 3.3 µm). Our quantitative analysis of the automated segmentation results from AION subject data revealed that the inner retinal layer thickness for the affected eye was 24.1 µm (21%) smaller on average than for the unaffected eye (P < 0.001), supporting the need for segmenting the layers separately. PMID:18815101

  2. 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. PMID:25046646

  3. 3D Image Processing of Two-Photon Microscopy Images Depicting Nanoprobes in Skin

    NASA Astrophysics Data System (ADS)

    Bongo, Andrew E.

    Choosing a deconvolution algorithm can be beneficial when imaging nanoprobes in skin by means of two-photon microscopy. By design, deconvolution algorithms can increase the signal to noise ratio of the raw images and thus make it easier to identify discrete, subresolution nanoprobes from blurry two-photon image data. This poses the favorable benefit of knowing more precise locations of nanoprobes inside skin. This thesis demonstrates how the Expectation-Maximization deconvolution algorithm (EM algorithm) can be applied to three-dimensional, two-photon images depicting quantum dot nanoprobes inside human skin. This was accomplished in part by devising a way to deliver nanoprobes inside skin by means of low frequency ultrasound. Many nanoprobes become sparsely scattered inside skin when using this nanoprobe delivery methodology. The scattered nanoprobes resulting from the nanoprobe delivery pose a unique benefit in acquiring an experimental point spread function of the imaging system. This in turn gives an accurate representation of the point spread function that can be used as an input to the EM algorithm. The methodology of utilizing the EM algorithm in this manner is presented.

  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. PMID:26399990

  5. A modular hierarchical approach to 3D electron microscopy image segmentation.

    PubMed

    Liu, Ting; Jones, Cory; Seyedhosseini, Mojtaba; Tasdizen, Tolga

    2014-04-15

    The study of neural circuit reconstruction, i.e., connectomics, is a challenging problem in neuroscience. Automated and semi-automated electron microscopy (EM) image analysis can be tremendously helpful for connectomics research. In this paper, we propose a fully automatic approach for intra-section segmentation and inter-section reconstruction of neurons using EM images. A hierarchical merge tree structure is built to represent multiple region hypotheses and supervised classification techniques are used to evaluate their potentials, based on which we resolve the merge tree with consistency constraints to acquire final intra-section segmentation. Then, we use a supervised learning based linking procedure for the inter-section neuron reconstruction. Also, we develop a semi-automatic method that utilizes the intermediate outputs of our automatic algorithm and achieves intra-segmentation with minimal user intervention. The experimental results show that our automatic method can achieve close-to-human intra-segmentation accuracy and state-of-the-art inter-section reconstruction accuracy. We also show that our semi-automatic method can further improve the intra-segmentation accuracy. PMID:24491638

  6. 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.

  7. Modeling of the general astigmatic Gaussian beam and its propagation through 3D optical systems.

    PubMed

    Kochkina, Evgenia; Wanner, Gudrun; Schmelzer, Dennis; Tröbs, Michael; Heinzel, Gerhard

    2013-08-20

    The paper introduces the complete model of the general astigmatic Gaussian beam as the most general case of the Gaussian beam in the fundamental mode. This includes the laws of propagation, reflection, and refraction as well as the equations for extracting from the complex-valued beam description its real-valued parameters, such as the beam spot radii and the radii of curvature of the wavefront. The suggested model is applicable to the case of an oblique incidence of the beam at any 3D surface that can be approximated by the second-order equation at the point of incidence. Thus it can be used in simulations of a large variety of 3D optical systems. The provided experimental validation of the model shows good agreement with simulations. PMID:24085008

  8. Architectures and algorithms for all-optical 3D signal processing

    NASA Astrophysics Data System (ADS)

    Giglmayr, Josef

    1999-07-01

    All-optical signal processing by >= 2D lightwave circuits (LCs) is (i) aimed to allow the (later) inclusion of the frequency domain and is (ii) subject to photonic integration and thus the architectural and algorithmic framework has to be prepared carefully. Much work has been done in >= 2D algebraic system theory/modern control theory which has been applied in the electronic field of signal and image processing. For the application to modeling, analysis and design of the proposed 3D lightwave circuits (LCs) some elements are needed to describe and evalute the system efficiency as the number of system states of 3D LCs increases dramatically with regard to the number of i/o. Several problems, arising throughput such an attempt, are made transparent and solutions are proposed.

  9. 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.

  10. Absolute 3D reconstruction of thin films topography in microfluidic channels by interference reflection microscopy.

    PubMed

    Huerre, A; Jullien, M-C; Theodoly, O; Valignat, M-P

    2016-03-01

    The travel of droplets, bubbles, vesicles, capsules, living cells or small organisms in microchannels is a hallmark in microfluidics applications. A full description of the dynamics of such objects requires a quantitative understanding of the complex hydrodynamic and interfacial interactions between objects and channel walls. In this paper, we present an interferometric method that allows absolute topographic reconstruction of the interspace between an object and channel walls for objects confined in microfluidic channels. Wide field microscopic imaging in reflection interference contrast mode (RICM) is directly performed at the bottom wall of microfluidic chips. Importantly, we show that the reflections at both the lower and upper surface of the microchannel have to be considered in the quantitative analysis of the optical signal. More precisely, the contribution of the reflection at the upper surface is weighted depending on the light coherence length and channel height. Using several wavelengths and illumination apertures, our method allows reconstructing the topography of thin films on channel walls in a range of 0-500 nm, with a precision as accurate as 2 nm for the thinnest films. A complete description of the protocol is exemplified for oil in water droplets travelling in channels of height 10-400 μm at a speed up to 5 mm s(-1). PMID:26830018

  11. 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

  12. 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

  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

  14. 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.

  15. 3D modeling for solving forward model of no-contact fluorescence diffuse optical tomography method

    NASA Astrophysics Data System (ADS)

    Nouizi, F.; Chabrier, R.; Torregrossa, M.; Poulet, P.

    2009-07-01

    This paper presents detailed computational aspects of a new 3D modeling for solving the direct problem in a no-contact time-resolved Fluorescent Diffuse Optical Tomography (FDOT) method that rely on near-infrared scattered and fluorescent photons to image the optical properties and distribution of fluorescent probes in small laboratory animals. An optical scanner allowing performing in-vivo measurements in no-contact scheme was built in our laboratory and is presented. We use the three-dimensional Finite Element Method (FEM) to solve the coupled diffusion equations of excitation and fluorescence photons in highly scattering objects. The computed results allowed yielding photon density maps and the temporal profiles of photons on the surface of the small animal. Our 3D modeling of propagation of photons in the void space between the surface of the object and the detectors allows calculating the quantity of photons reaching the optodes. Simulations were carried-out on two test objects: a resin cylinder and a mouse phantom. The results demonstrate the potential applications of the method to pre-clinical imaging.

  16. Optical flow 3D segmentation and interpretation: a variational method with active curve evolution and level sets.

    PubMed

    Mitiche, Amar; Sekkati, Hicham

    2006-11-01

    This study investigates a variational, active curve evolution method for dense three-dimentional (3D) segmentation and interpretation of optical flow in an image sequence of a scene containing moving rigid objects viewed by a possibly moving camera. This method jointly performs 3D motion segmentation, 3D interpretation (recovery of 3D structure and motion), and optical flow estimation. The objective functional contains two data terms for each segmentation region, one based on the motion-only equation which relates the essential parameters of 3D rigid body motion to optical flow, and the other on the Horn and Schunck optical flow constraint. It also contains two regularization terms for each region, one for optical flow, the other for the region boundary. The necessary conditions for a minimum of the functional result in concurrent 3D-motion segmentation, by active curve evolution via level sets, and linear estimation of each region essential parameters and optical flow. Subsequently, the screw of 3D motion and regularized relative depth are recovered analytically for each region from the estimated essential parameters and optical flow. Examples are provided which verify the method and its implementation. PMID:17063686

  17. 3D optical measurement of relative displacement for the tuberosities in the shoulder prosthesis

    NASA Astrophysics Data System (ADS)

    Yao, XueFeng; Meng, LiBo; Yu, LiuPing; Zhu, YiMing; Jiang, ChunYan

    2011-04-01

    In this paper, a binocular 3-D computer vision measurement system is used to measure the relative displacement for the greater and lesser tuberosities in the shoulder prosthesis. The basic principles of binocular optical measurement are introduced in detail, and the loading apparatus is designed for external rotation and anteflexion of the shoulder prosthesis. Both the motion of external rotation and anteflexion of the shoulder are measured, and the corresponding displacement values for the greater and lesser tuberosities are extracted. These results will play an important role in evaluating the stability of humeral tuberosity in the shoulder prosthesis.

  18. New light sources and sensors for active optical 3D inspection

    NASA Astrophysics Data System (ADS)

    Osten, Wolfgang; Jueptner, Werner P. O.

    1999-11-01

    The implementation of active processing strategies in optical 3D-inspection needs the availability of flexible hardware solutions. The system components illumination and sensor/detector are actively involved in the processing chain by a feedback loop that is controlled by the evaluation process. Therefore this article deals with new light sources and sensor which appeared recently on the market and can be applied successfully for the implementation of active processing principles. Some applications where such new components are used to implement an active measurement strategy are presented.

  19. 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

  20. Shot noise limit of the optical 3D measurement methods for smooth surfaces

    NASA Astrophysics Data System (ADS)

    Pavliček, Pavel; Pech, Miroslav

    2016-03-01

    The measurement uncertainty of optical 3D measurement methods for smooth surfaces caused by shot noise is investigated. The shot noise is a fundamental property of the quantum nature of light. If all noise sources are eliminated, the shot noise represents the ultimate limit of the measurement uncertainty. The measurement uncertainty is calculated for several simple model methods. The analysis shows that the measurement uncertainty depends on the wavelength of used light, the number of photons used for the measurement, and on a factor that is connected with the geometric arrangement of the measurement setup.

  1. 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.

  2. DLP/DSP-based optical 3D sensors for the mass market in industrial metrology and life sciences

    NASA Astrophysics Data System (ADS)

    Frankowski, G.; Hainich, R.

    2011-03-01

    GFM has developed and constructed DLP-based optical 3D measuring devices based on structured light illumination. Over the years the devices have been used in industrial metrology and life sciences for different 3D measuring tasks. This lecture will discuss integration of DLP Pico technology and DSP technology from Texas Instruments for mass market optical 3D sensors. In comparison to existing mass market laser triangulation sensors, the new 3D sensors provide a full-field measurement of up to a million points in less than a second. The lecture will further discuss different fields of application and advantages of the new generation of 3D sensors for: OEM application in industrial measuring and inspection; 3D metrology in industry, life sciences and biometrics, and industrial image processing.

  3. 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).

  4. 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.

  5. Optical fiber sensor system for oil contamination measurement based on 3D fluorescence spectrum parameterization

    NASA Astrophysics Data System (ADS)

    Shang, Liping; Shi, Jinshan

    2000-10-01

    In recent years oil contamination in water is more serious and destroys the mode of life and relation to water body environments. Excitation fluorescence method is one of the main approaches to monitor oil contamination on line. But average intensity of oil fluorescence only indicates its density, not indicates the type of contamination oil. Two-dimensional fluorescence spectrum is more difficult to determine the kind of oil, because the different oil has fluorescence spectrum overlapping to a great extent. In this paper, the 3D fluorescence spectrum parameterization is introduced. It can extract several characteristic parameters to measure the kid of oil to be measured. A prototype of optical fiber 3D fluorescence spectrum meter we developed carries out the identification of different oil types, such as crude oil, diesel oil and kerosene. The experiment arrangement conceived to measure pulse xenon lamp induced of oil component in water. The experiment results state clearly that the 3D fluorescence spectrum parameterization and software are successful to measure oil density and identify the type of oil in situ.

  6. 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.

  7. 3D multiple optical tweezers based on time-shared scanning with a fast focus tunable lens

    NASA Astrophysics Data System (ADS)

    Tanaka, Yoshio

    2013-02-01

    Three-dimensional controlled manipulation of individual micro-objects requires multiple optical tweezers that can be independently controlled in a 3D working space with high spatiotemporal resolution. Here, the author presents 3D multiple optical tweezers based on a time-shared scanning technique with an electrically focus tunable lens for axial steering and a two-axis steering mirror for lateral steering. Four typical examples of 3D controlled manipulation, including the rotation of a single bead on its axis, are demonstrated in real time. The optical system design and the control method are also described.

  8. Direct Determination of 3D Distribution of Elemental Composition in Single Semiconductor Nanoislands by Scanning Auger Microscopy.

    PubMed

    Ponomaryov, Semyon S; Yukhymchuk, Volodymyr O; Lytvyn, Peter M; Valakh, Mykhailo Ya

    2016-12-01

    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.%. PMID:26909783

  9. Low-temperature post-deposition annealing investigation for 3D charge trap flash memory by Kelvin probe force microscopy

    NASA Astrophysics Data System (ADS)

    Huo, Zongliang; Jin, Lei; Han, Yulong; Li, Xinkai; Ye, Tianchun; Liu, Ming

    2015-01-01

    The influence of post-deposition annealing (PDA) temperature condition on charge distribution behavior of HfO2 thin films was systematically investigated by various-temperature Kelvin probe force microscopy technology. Contact potential difference profiles demonstrated that charge storage capability shrinks with decreasing annealing temperature from 1,000 to 500 °C and lower. Compared to 1,000 °C PDA, it was found that 500 °C PDA causes deeper effective trap energy level, suppresses lateral charge spreading, and improves the retention characteristics. It is concluded that low-temperature PDA can be adopted in 3D HfO2-based charge trap flash memory to improve the thermal treatment compatibility of the bottom peripheral logic and upper memory arrays.

  10. 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

  11. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

    DOE PAGESBeta

    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. PMID:27375939

  13. 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.%.

  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. 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.

  16. 3D imaging of biofilms on implants by detection of scattered light with a scanning laser optical tomograph

    PubMed Central

    Heidrich, Marko; Kühnel, Mark P.; Kellner, Manuela; Lorbeer, Raoul-Amadeus; Lange, Tineke; Winkel, Andreas; Stiesch, Meike; Meyer, Heiko; Heisterkamp, Alexander

    2011-01-01

    Biofilms – communities of microorganisms attached to surfaces – are a constant threat for long-term success in modern implantology. The application of laser scanning microscopy (LSM) has increased the knowledge about microscopic properties of biofilms, whereas a 3D imaging technique for the large scale visualization of bacterial growth and migration on curved and non-transparent surfaces is not realized so far. Towards this goal, we built a scanning laser optical tomography (SLOT) setup detecting scattered laser light to image biofilm on dental implant surfaces. SLOT enables the visualization of living biofilms in 3D by detecting the wavelength-dependent absorption of non-fluorescent stains like e.g. reduced triphenyltetrazolium chloride (TTC) accumulated within metabolically active bacterial cells. Thus, the presented system allows the large scale investigation of vital biofilm structure and in vitro development on cylindrical and non-transparent objects without the need for fluorescent vital staining. We suggest SLOT to be a valuable tool for the structural and volumetric investigation of biofilm formation on implants with sizes up to several millimeters. PMID:22076261

  17. 3D visualization of tissue microstructures using optical coherence tomography needle probes

    NASA Astrophysics Data System (ADS)

    Kirk, Rodney W.; McLaughlin, Robert A.; Quirk, Bryden C.; Curatolo, Andrea; Sampson, David D.

    2011-05-01

    Optical coherence tomography (OCT) needle probes use miniaturized focusing optics encased in a hypodermic needle. Needle probes can scan areas of the body that are too deep to be imaged by other OCT systems. This paper presents an OCT needle probe-based system that is capable of acquiring three-dimensional scans of tissue structures. The needle can be guided to a target area and scans acquired by rotating and pulling-back the probe. The system is demonstrated using ex vivo human lymph node and sheep lung samples. Multiplanar reconstructions are shown of both samples, as well as the first published 3D volume rendering of lung tissue acquired with an OCT needle probe.

  18. The Quantum Dynamics of a Dilute Gas in a 3D BCC Optical Lattice

    NASA Astrophysics Data System (ADS)

    Reichl, Linda; Boretz, Yingyue

    2015-03-01

    The classical and quantum dynamics of a dilute gas of rubidium atoms, in a 3D body-centered cubic optical lattice, is studied for a range of polarizations of the laser beams forming the lattice. The relative polarization of the lasers determines the the structure of the potential energy seen by the rubidium atoms. If three pairs of in-phase mutually perpendicular laser beams, with the same wavelength, form the lattice, only a limited range of possible couplings can be realized in the lab. We have determined the band structure of the BCC optical lattice for all theoretically possible couplings, and find that the band structure for lattices realizable in the lab, differs significantly from that expected for a BCC crystal. As coupling is increased, the lattice becomes increasingly chaotic and it becomes possible to produce band structure that has qualitative similarity to a BCC. Welch Foundation

  19. 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.

  20. Implementation of 3D prostrate ring-scanning mechanism for NIR diffuse optical imaging phantom validation

    NASA Astrophysics Data System (ADS)

    Yu, Jhao-Ming; Chen, Liang-Yu; Pan, Min-Cheng; Hsu, Ya-Fen; Pan, Min-Chun

    2015-03-01

    Diffuse optical imaging (DOI) providing functional information of tissues has drawn great attention for the last two decades. Near infrared (NIR) DOI systems composed of scanning bench, opt-electrical measurement module, system control, and data processing and image reconstruction schemes are developed for the screening and diagnosis of breast tumors. Mostly, the scanning bench belonging to fixed source-and-detector configuration limits computed image resolution to an extent. To cope with the issue, we propose, design and implement a 3D prostrate ring-scanning equipment for NIR DOI with flexible combinations of illumination and detection, and with the function of radial, circular and vertical movement without hard compression of breast tissue like the imaging system using or incorporating with X-ray mammographic bench. Especially, a rotation-sliding-and-moving mechanism was designed for the guidance of source- and detection-channel movement. Following the previous justification for synthesized image reconstruction, in the paper the validation using varied phantoms is further conducted and 3D image reconstruction for their absorption and scattering coefficients is illustrated through the computation of our in-house coded schemes. The source and detection NIR data are acquired to reconstruct the 3D images through the operation of scanning bench in the movement of vertical, radial and circular directions. Rather than the fixed configuration, the addressed screening/diagnosing equipment has the flexibility for optical-channel expansion with a compromise among construction cost, operation time, and spatial resolution of reconstructed μa and μs' images.

  1. 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.

  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. 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

  4. How effective can optical-CT 3D dosimetry be without refractive fluid matching?

    NASA Astrophysics Data System (ADS)

    Rankine, L.; Oldham

    2013-06-01

    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.

  5. 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. PMID:26529743

  6. 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.

  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. 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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. Enhancing 3-D cell structures in confocal and STED microscopy: a joint model for interpolation, deblurring and anisotropic smoothing

    NASA Astrophysics Data System (ADS)

    Persch, Nico; Elhayek, Ahmed; Welk, Martin; Bruhn, Andrés; Grewenig, Sven; Böse, Katharina; Kraegeloh, Annette; Weickert, Joachim

    2013-12-01

    This paper proposes an advanced image enhancement method that is specifically tailored towards 3-D confocal and STED microscopy imagery. Our approach unifies image denoising, deblurring and interpolation in one joint method to handle the typical weaknesses of these advanced microscopy techniques: out-of-focus blur, Poisson noise and low axial resolution. In detail, we propose the combination of (i) Richardson-Lucy deconvolution, (ii) image restoration and (iii) anisotropic inpainting in one single scheme. To this end, we develop a novel PDE-based model that realizes these three ideas. First we consider a basic variational image restoration functional that is turned into a joint interpolation scheme by extending the regularization domain. Next, we integrate the variational representation of Richardson-Lucy deconvolution into our model, and illustrate its relation to Poisson distributed noise. In the following step, we supplement the components of our model with sub-quadratic penalization strategies that increase the robustness of the overall method. Finally, we consider the associated minimality conditions, where we exchange the occurring scalar-valued diffusivity function by a so-called diffusion tensor. This leads to an anisotropic regularization that is aligned with structures in the evolving image. As a further contribution of this paper, we propose a more efficient and faster semi-implicit iteration scheme that also increases the stability. Our experiments on real data sets demonstrate that this joint model achieves a superior reconstruction quality of the recorded cell.

  14. 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

  15. 3D soliton-like bullets in nonlinear optics and Bose-Einstein condensates

    NASA Astrophysics Data System (ADS)

    Belyaeva, T. L.; Hasegawa, Akira; Kovachev, L. M.; Serkin, V. N.

    2010-10-01

    Mathematical similarities and parallels between two different physical objects, optical solitons and matter-wave solitons, both described by similar mathematical models: the nonlinear Schrodinger equation (NLSE) and the Gross-Pitaevskii equation (GPE) model, open the possibility to study both systems in parallel and because of the obvious complexity of experiments with matter-wave solitons, offer outstanding possibilities in studies of BEC system by performing experiments in the nonlinear optical system and vise versa. In this report we briefly overview recent theoretical studies of the existence and stability of 3D solitons. With contributions from major groups who have pioneered research in this field, the report describes the historical development of the subject, provides a background to the associated nonlinear optical processes, the generation mechanisms of soliton bullets. The main features of nonautonomous matter-wave solitons near the Feshbach resonance with continuously tuned scattering length are investigated. We focus on the most physically important situations where the applied magnetic field is varying in time linearly and periodically. In nonlinear optical applications, this kind of periodic graded-index nonlinear structure with alternating waveguiding and antiwaveguiding segments can be used to simulate different and complicated processes in the total scenario of matterwave soliton bullets generation.

  16. 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.

  17. Coherent nonlinear optical imaging: beyond fluorescence microscopy.

    PubMed

    Min, Wei; Freudiger, Christian W; Lu, Sijia; Xie, X Sunney

    2011-01-01

    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 excited-state absorption, stimulated emission, and ground-state depletion), provides new image contrasts for nonfluorescent species. Thanks to the high-frequency modulation transfer scheme, these imaging techniques 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 and excitation and detection schemes, as well as exemplary biomedical applications of this emerging class of molecular imaging techniques. PMID:21453061

  18. 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

  19. 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

  20. 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

  1. 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.

  2. 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

  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. Hybrid system of optics and computer for 3-D object recognition

    NASA Astrophysics Data System (ADS)

    Li, Qun Z.; Miao, Peng C.; He, Anzhi

    1992-03-01

    In this paper, a hybrid system of optics and computer for 3D object recognition is presented. The system consists of a Twyman-Green interferometer, a He-Ne laser, a computer, a TV camera, and an image processor. The structured light produced by a Twyman-Green interferometer is split in and illuminates objects in two directions at the same time. Moire contour is formed on the surface of object. In order to delete unwanted patterns in moire contour, we don't utilize the moire contour on the surface of object. We place a TV camera in the middle of the angle between two illuminating directions and take two groups of deformed fringes on the surface of objects. Two groups of deformed fringes are processed using the digital image processing system controlled and operated by XOR logic in the computer, moire fringes are then extracted from the complicated environment. 3D coordinates of points of the object are obtained after moire fringe is followed, and points belonging to the same fringe are given the same altitude. The object is described by its projected drawings in three coordinate planes. The projected drawings in three coordinate planes of the known objects are stored in the library of judgment. The object can be recognized by inquiring the library of judgment.

  5. 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.

  6. Optically directed molecular transport and 3D isoelectric positioning of amphoteric biomolecules

    SciTech Connect

    Hafeman, Dean G.; Harkins, James B.; WitkowskiII, Charles E.; Lewis, Nathan S.; Brown, Gilbert M; Warmack, Robert J Bruce; Thundat, Thomas George

    2006-01-01

    We demonstrate the formation of charged molecular packets and their transport within optically created electrical force-field traps in a pH-buffered electrolyte. We call this process photoelectrophoretic localization and transport (PELT). The electrolyte is in contact with a photoconductive semiconductor electrode and a counterelectrode that are connected through an external circuit. A light beam directed to coordinates on the photoconductive electrode surface produces a photocurrent within the circuit and electrolyte. Within the electrolyte, the photocurrent creates localized force-field traps centered at the illuminated coordinates. Charged molecules, including polypeptides and proteins, electrophoretically accumulate into the traps and subsequently can be transported in the electrolyte by moving the traps over the photoconductive electrode in response to movement of the light beam. The molecules in a single trap can be divided into aliquots, and the aliquots can be directed along multiple routes simultaneously by using multiple light beams. This photoelectrophoretic transport of charged molecules by PELT resembles the electrostatic transport of electrons within force-field wells of solid-state charge-coupled devices. The molecules, however, travel in a liquid electrolyte rather than a solid. Furthermore, we have used PELT to position amphoteric biomolecules in three dimensions. A 3D pH gradient was created in an electrolyte medium by controlling the illumination position on a photoconductive anode where protons were generated electrolytically. Photoelectrophoretic transport of amphoteric molecules through the pH gradient resulted in accumulation of the molecules at their apparent 3D isoelectric coordinates in the medium.

  7. 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%. PMID:26342493

  8. 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.

  9. Optical projection tomography as a tool for 3D imaging of hydrogels

    PubMed Central

    Figueiras, Edite; Soto, Ana M.; Jesus, Danilo; Lehti, M.; Koivisto, J.; Parraga, J. E.; Silva-Correia, J.; Oliveira, J. M.; Reis, R. L.; Kellomäki, M.; Hyttinen, J.

    2014-01-01

    An Optical Projection Tomography (OPT) system was developed and optimized to image 3D tissue engineered products based in hydrogels. We develop pre-reconstruction algorithms to get the best result from the reconstruction procedure, which include correction of the illumination and determination of sample center of rotation (CoR). Existing methods for CoR determination based on the detection of the maximum variance of reconstructed slices failed, so we develop a new CoR search method based in the detection of the variance sharpest local maximum. We show the capabilities of the system to give quantitative information of different types of hydrogels that may be useful in its characterization. PMID:25360363

  10. 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

  11. 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.

  12. 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

  13. 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

  14. 3D image restoration for confocal microscopy: toward a wavelet deconvolution for the study of complex biological structures

    NASA Astrophysics Data System (ADS)

    Boutet de Monvel, Jacques; Le Calvez, Sophie; Ulfendahl, Mats

    2000-05-01

    Image restoration algorithms provide efficient tools for recovering part of the information lost in the imaging process of a microscope. We describe recent progress in the application of deconvolution to confocal microscopy. The point spread function of a Biorad-MRC1024 confocal microscope was measured under various imaging conditions, and used to process 3D-confocal images acquired in an intact preparation of the inner ear developed at Karolinska Institutet. Using these experiments we investigate the application of denoising methods based on wavelet analysis as a natural regularization of the deconvolution process. Within the Bayesian approach to image restoration, we compare wavelet denoising with the use of a maximum entropy constraint as another natural regularization method. Numerical experiments performed with test images show a clear advantage of the wavelet denoising approach, allowing to `cool down' the image with respect to the signal, while suppressing much of the fine-scale artifacts appearing during deconvolution due to the presence of noise, incomplete knowledge of the point spread function, or undersampling problems. We further describe a natural development of this approach, which consists of performing the Bayesian inference directly in the wavelet domain.

  15. A computational 3D model for reconstruction of neural stem cells in bright-field time-lapse microscopy

    NASA Astrophysics Data System (ADS)

    Degerman, J.; Winterfors, E.; Faijerson, J.; Gustavsson, T.

    2007-02-01

    This paper describes a computational model for image formation of in-vitro adult hippocampal progenitor (AHP) cells, in bright-field time-lapse microscopy. Although this microscopymodality barely generates sufficient contrast for imaging translucent cells, we show that by using a stack of defocused image slices it is possible to extract position and shape of spherically shaped specimens, such as the AHP cells. This inverse problem was solved by modeling the physical objects and image formation system, and using an iterative nonlinear optimization algorithm to minimize the difference between the reconstructed and measured image stack. By assuming that the position and shape of the cells do not change significantly between two time instances, we can optimize these parameters using the previous time instance in a Bayesian estimation approach. The 3D reconstruction algorithm settings, such as focal sampling distance, and PSF, were calibrated using latex spheres of known size and refractive index. By using the residual between reconstructed and measured image intensities, we computed a peak signal-to-noise ratio (PSNR) to 28 dB for the sphere stack. A biological specimen analysis was done using an AHP cell, where reconstruction PSNR was 28 dB as well. The cell was immuno-histochemically stained and scanned in a confocal microscope, in order to compare our cell model to a ground truth. After convergence the modelled cell volume had an error of less than one percent.

  16. Association of intracellular and synaptic organization in cochlear inner hair cells revealed by 3D electron microscopy

    PubMed Central

    Bullen, Anwen; West, Timothy; Moores, Carolyn; Ashmore, Jonathan; Fleck, Roland A.; MacLellan-Gibson, Kirsty; Forge, Andrew

    2015-01-01

    ABSTRACT The ways in which cell architecture is modelled to meet cell function is a poorly understood facet of cell biology. To address this question, we have studied the cytoarchitecture of a cell with highly specialised organisation, the cochlear inner hair cell (IHC), using multiple hierarchies of three-dimensional (3D) electron microscopy analyses. We show that synaptic terminal distribution on the IHC surface correlates with cell shape, and the distribution of a highly organised network of membranes and mitochondria encompassing the infranuclear region of the cell. This network is juxtaposed to a population of small vesicles, which represents a potential new source of neurotransmitter vesicles for replenishment of the synapses. Structural linkages between organelles that underlie this organisation were identified by high-resolution imaging. Taken together, these results describe a cell-encompassing network of membranes and mitochondria present in IHCs that support efficient coding and transmission of auditory signals. Such techniques also have the potential for clarifying functionally specialised cytoarchitecture of other cell types. PMID:26045447

  17. Hyperlens-array-implemented optical microscopy

    NASA Astrophysics Data System (ADS)

    Iwanaga, Masanobu

    2014-08-01

    Limit of resolution of conventional optical microscopes has never reached below 100 nm under visible light illumination. We show that numerically designed high-transmittance hyperlens array (HLA) is implemented in an optical microscope and works in practice for achieving one-shot-recording optical images of in-situ placed objects with sub 50 nm resolution in lateral direction. Direct resolution test employing well-defined nanopatterns proves that the HLA-implemented imaging is super-resolution optical microscopy, which works even under nW/mm2 visible illumination for objects. The HLA implementation makes the resolution of conventional microscopes one-scale higher, leading to the 1/10 illumination wavelength range, that is, mesoscopic range.

  18. 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

  19. 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. PMID:11015933

  20. 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.

  1. Virtual k-Space Modulation Optical Microscopy.

    PubMed

    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. PMID:27447529

  2. Pulse front adaptive optics in multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Sun, B.; Salter, P. S.; Booth, M. J.

    2016-03-01

    The accurate focusing of ultrashort laser pulses is extremely important in multiphoton microscopy. Using adaptive optics to manipulate the incident ultrafast beam in either the spectral or spatial domain can introduce significant benefits when imaging. Here we introduce pulse front adaptive optics: manipulating an ultrashort pulse in both the spatial and temporal domains. A deformable mirror and a spatial light modulator are operated in concert to modify contours of constant intensity in space and time within an ultrashort pulse. Through adaptive control of the pulse front, we demonstrate an enhancement in the measured fluorescence from a two photon microscope.

  3. Multiple scattering in optical coherence microscopy.

    PubMed

    Yadlowsky, M J; Schmitt, J M; Bonner, R F

    1995-09-01

    We show that the multiple-scatter rejection provided by optical coherence microscopy (low-coherence interferometry) can be incomplete in optically turbid media and that multiple scattering manifests itself in two distinct ways. Multiple small-angle scattering results in an effective probe field that is stronger than expected from a first-order beam extinction model, but that contains a distorted wave front that enhances the apparent reflectance of small structures relative to those that are larger than the unscattered incident beam. Multiple wide-angle scattering produces a broad diffuse haze that reduces the contrast of subsequent features. PMID:21060400

  4. 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.

  5. 3D imaging of dental hard tissues with Fourier domain optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Chen, Yueli L.; Yang, Yi; Ma, Jing; Yan, Jun; Shou, Yuanxin; Wang, Tianheng; Ramesh, Aruna; Zhao, Jing; Zhu, Quing

    2011-03-01

    A fiber optical coherence tomography (OCT) probe is used for three dimensional dental imaging. The probe has a lightweight miniaturized design with a size of a pen to facilitate clinic in vivo diagnostics. The probe is interfaced with a swept-source / Fourier domain optical coherence tomography at 20K axial scanning rate. The tooth samples were scanned from occlusal, buccal, lingual, mesial, and distal orientations. Three dimensional imaging covers tooth surface area up to 10 mm x 10 mm with a depth about 5 mm, where a majority of caries affection occurs. OCT image provides better resolution and contrast compared to gold standard dental radiography (X-ray). In particular, the technology is well suited for occlusal caries detection. This is complementary to X-ray as occlusal caries affection is difficult to be detected due to the X-ray projectile scan geometry. The 3D topology of occlusal surface as well as the dentin-enamel junction (DEJ) surface inside the tooth can be visualized. The lesion area appears with much stronger back scattering signal intensity.

  6. Near-field optical microscopy nanoarray

    NASA Astrophysics Data System (ADS)

    Semin, David J.; Ambrose, W. Patrick; Goodwin, Peter M.; Wendt, Joel R.; Keller, Richard A.

    1997-04-01

    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 108 apertures were fabricated by electron beam lithography and reactive ion etching. The nanoarrays were characterized by atomic force microscopy and scanning electron microscopy. In this paper we utilize these nanoarrays in a laser-illuminated microscope with parallel detection on a charge-coupled device. Detection of B-phycoerythrin 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 micrometers ) within seconds.

  7. 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.

  8. Correlative super-resolution fluorescence microscopy combined with optical coherence microscopy

    NASA Astrophysics Data System (ADS)

    Kim, Sungho; Kim, Gyeong Tae; Jang, Soohyun; Shim, Sang-Hee; Bae, Sung Chul

    2015-03-01

    Recent development of super-resolution fluorescence imaging technique such as stochastic optical reconstruction microscopy (STORM) and photoactived localization microscope (PALM) has brought us beyond the diffraction limits. It allows numerous opportunities in biology because vast amount of formerly obscured molecular structures, due to lack of spatial resolution, now can be directly observed. A drawback of fluorescence imaging, however, is that it lacks complete structural information. For this reason, we have developed a super-resolution multimodal imaging system based on STORM and full-field optical coherence microscopy (FF-OCM). FF-OCM is a type of interferometry systems based on a broadband light source and a bulk Michelson interferometer, which provides label-free and non-invasive visualization of biological samples. The integration between the two systems is simple because both systems use a wide-field illumination scheme and a conventional microscope. This combined imaging system gives us both functional information at a molecular level (~20nm) and structural information at the sub-cellular level (~1μm). For thick samples such as tissue slices, while FF-OCM is readily capable of imaging the 3D architecture, STORM suffer from aberrations and high background fluorescence that substantially degrade the resolution. In order to correct the aberrations in thick tissues, we employed an adaptive optics system in the detection path of the STORM microscope. We used our multimodal system to obtain images on brain tissue samples with structural and functional information.

  9. 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.

  10. Optically directed molecular transport and 3D isoelectric positioning of amphoteric biomolecules

    PubMed Central

    Hafeman, Dean G.; Harkins, James B.; Witkowski, Charles E.; Lewis, Nathan S.; Warmack, Robert J.; Brown, Gilbert M.; Thundat, Thomas

    2006-01-01

    We demonstrate the formation of charged molecular packets and their transport within optically created electrical force-field traps in a pH-buffered electrolyte. We call this process photoelectrophoretic localization and transport (PELT). The electrolyte is in contact with a photoconductive semiconductor electrode and a counterelectrode that are connected through an external circuit. A light beam directed to coordinates on the photoconductive electrode surface produces a photocurrent within the circuit and electrolyte. Within the electrolyte, the photocurrent creates localized force-field traps centered at the illuminated coordinates. Charged molecules, including polypeptides and proteins, electrophoretically accumulate into the traps and subsequently can be transported in the electrolyte by moving the traps over the photoconductive electrode in response to movement of the light beam. The molecules in a single trap can be divided into aliquots, and the aliquots can be directed along multiple routes simultaneously by using multiple light beams. This photoelectrophoretic transport of charged molecules by PELT resembles the electrostatic transport of electrons within force-field wells of solid-state charge-coupled devices. The molecules, however, travel in a liquid electrolyte rather than a solid. Furthermore, we have used PELT to position amphoteric biomolecules in three dimensions. A 3D pH gradient was created in an electrolyte medium by controlling the illumination position on a photoconductive anode where protons were generated electrolytically. Photoelectrophoretic transport of amphoteric molecules through the pH gradient resulted in accumulation of the molecules at their apparent 3D isoelectric coordinates in the medium. PMID:16618926

  11. 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

  12. Multimodal photoacoustic and optical coherence tomography scanner using an all optical detection scheme for 3D morphological skin imaging

    PubMed Central

    Zhang, Edward Z.; Povazay, Boris; Laufer, Jan; Alex, Aneesh; Hofer, Bernd; Pedley, Barbara; Glittenberg, Carl; Treeby, Bradley; Cox, Ben; Beard, Paul; Drexler, Wolfgang

    2011-01-01

    A noninvasive, multimodal photoacoustic and optical coherence tomography (PAT/OCT) scanner for three-dimensional in vivo (3D) skin imaging is described. The system employs an integrated, all optical detection scheme for both modalities in backward mode utilizing a shared 2D optical scanner with a field-of-view of ~13 × 13 mm2. The photoacoustic waves were detected using a Fabry Perot polymer film ultrasound sensor placed on the surface of the skin. The sensor is transparent in the spectral range 590-1200 nm. This permits the photoacoustic excitation beam (670-680 nm) and the OCT probe beam (1050 nm) to be transmitted through the sensor head and into the underlying tissue thus providing a backward mode imaging configuration. The respective OCT and PAT axial resolutions were 8 and 20 µm and the lateral resolutions were 18 and 50-100 µm. The system provides greater penetration depth than previous combined PA/OCT devices due to the longer wavelength of the OCT beam (1050 nm rather than 829-870 nm) and by operating in the tomographic rather than the optical resolution mode of photoacoustic imaging. Three-dimensional in vivo images of the vasculature and the surrounding tissue micro-morphology in murine and human skin were acquired. These studies demonstrated the complementary contrast and tissue information provided by each modality for high-resolution 3D imaging of vascular structures to depths of up to 5 mm. Potential applications include characterizing skin conditions such as tumors, vascular lesions, soft tissue damage such as burns and wounds, inflammatory conditions such as dermatitis and other superficial tissue abnormalities. PMID:21833358

  13. In vivo tissue has non-linear rheological behavior distinct from 3D biomimetic hydrogels, as determined by AMOTIV microscopy.

    PubMed

    Blehm, Benjamin H; Devine, Alexus; Staunton, Jack R; Tanner, Kandice

    2016-03-01

    Variation in matrix elasticity has been shown to determine cell fate in both differentiation and development of malignant phenotype. The tissue microenvironment provides complex biochemical and biophysical signals in part due to the architectural heterogeneities found in extracellular matrices (ECMs). Three dimensional cell cultures can partially mimic in vivo tissue architecture, but to truly understand the role of viscoelasticity on cell fate, we must first determine in vivo tissue mechanical properties to improve in vitro models. We employed Active Microrheology by Optical Trapping InVivo (AMOTIV), using in situ calibration to measure in vivo zebrafish tissue mechanics. Previously used trap calibration methods overestimate complex moduli by ∼ 2-20 fold compared to AMOTIV. Applying differential microscale stresses and strains showed that hyaluronic acid (HA) gels display semi-flexible polymer behavior, while laminin-rich ECM hydrogels display flexible polymer behavior. In contrast, zebrafish tissues displayed different moduli at different stresses, with higher power law exponents at lower stresses, indicating that living tissue has greater stress dependence than the 3D hydrogels examined. To our knowledge, this work is the first vertebrate tissue rheological characterization performed in vivo. Our fundamental observations are important for the development and refinement of in vitro platforms. PMID:26773661

  14. 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

  15. 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

  16. 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

  17. Distributed microscopy: toward a 3D computer-graphic-based multiuser microscopic manipulation, imaging, and measurement system

    NASA Astrophysics Data System (ADS)

    Sulzmann, Armin; Carlier, Jerome; Jacot, Jacques

    1996-10-01

    The aim of this project is to telecontrol the movements in 3D-space of a microscope in order to manipulate and measure microsystems or micro parts aided by multi-user virtual reality (VR) environments. Presently microsystems are gaining in interest. Microsystems are small, independent modules, incorporating various functions, such as electronic, micro mechanical, data processing, optical, chemical, medical and biological functions. Though improving the manufacturing technologies, the measurement of the small structures to insure the quality of the process is a key information for the development. So far to measure the micro structures strong microscopes are needed. The use of highly magnifying computerized microscopes is expensive. To insure high quality measurements and distribute the acquired information to multi-user our proposed system is divided into three parts: the virtual reality microscopic environment (VRME)-based user-interface on a SGI workstation to prepare the manipulations and measurements. Secondly the computerized light microscope with the vision system inspecting the scene and getting the images of the specimen. Newly developed vision algorithms are used to analyze micro structures in the scene corresponding to the known a priori model. This vision is extracting position and shape of the objects and then transmitted as feedback to the user of the VRME-system to update his virtual environment. The internet demon is the third part of the system and distributes the information about the position of the micro structures, their shape and the images to the connected users who themselves may interact with the microscope (turn and displace the specimen on the back of a moving platform, or adding their structures to the scene and compare). The key idea behind our project VRME is to use the intuitiveness and the 3D visualization of VR environments coupled with a vision system to perform measurements of micro structures at a high accuracy. The direct

  18. 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

  19. 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.

  20. Exact surface registration of retinal surfaces from 3-D optical coherence tomography images.

    PubMed

    Lee, Sieun; Lebed, Evgeniy; Sarunic, Marinko V; Beg, Mirza Faisal

    2015-02-01

    Nonrigid registration of optical coherence tomography (OCT) images is an important problem in studying eye diseases, evaluating the effect of pharmaceuticals in treating vision loss, and performing group-wise cross-sectional analysis. High dimensional nonrigid registration algorithms required for cross-sectional and longitudinal analysis are still being developed for accurate registration of OCT image volumes, with the speckle noise in images presenting a challenge for registration. Development of algorithms for segmentation of OCT images to generate surface models of retinal layers has advanced considerably and several algorithms are now available that can segment retinal OCT images into constituent retinal surfaces. Important morphometric measurements can be extracted if accurate surface registration algorithm for registering retinal surfaces onto corresponding template surfaces were available. In this paper, we present a novel method to perform multiple and simultaneous retinal surface registration, targeted to registering surfaces extracted from ocular volumetric OCT images. This enables a point-to-point correspondence (homology) between template and subject surfaces, allowing for a direct, vertex-wise comparison of morphometric measurements across subject groups. We demonstrate that this approach can be used to localize and analyze regional changes in choroidal and nerve fiber layer thickness among healthy and glaucomatous subjects, allowing for cross-sectional population wise analysis. We also demonstrate the method's ability to track longitudinal changes in optic nerve head morphometry, allowing for within-individual tracking of morphometric changes. This method can also, in the future, be used as a precursor to 3-D OCT image registration to better initialize nonrigid image registration algorithms closer to the desired solution. PMID:25312906

  1. 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.

  2. Parallel robot for micro assembly with integrated innovative optical 3D-sensor

    NASA Astrophysics Data System (ADS)

    Hesselbach, Juergen; Ispas, Diana; Pokar, Gero; Soetebier, Sven; Tutsch, Rainer

    2002-10-01

    Recent advances in the fields of MEMS and MOEMS often require precise assembly of very small parts with an accuracy of a few microns. In order to meet this demand, a new approach using a robot based on parallel mechanisms in combination with a novel 3D-vision system has been chosen. The planar parallel robot structure with 2 DOF provides a high resolution in the XY-plane. It carries two additional serial axes for linear and rotational movement in/about z direction. In order to achieve high precision as well as good dynamic capabilities, the drive concept for the parallel (main) axes incorporates air bearings in combination with a linear electric servo motors. High accuracy position feedback is provided by optical encoders with a resolution of 0.1 μm. To allow for visualization and visual control of assembly processes, a camera module fits into the hollow tool head. It consists of a miniature CCD camera and a light source. In addition a modular gripper support is integrated into the tool head. To increase the accuracy a control loop based on an optoelectronic sensor will be implemented. As a result of an in-depth analysis of different approaches a photogrammetric system using one single camera and special beam-splitting optics was chosen. A pattern of elliptical marks is applied to the surfaces of workpiece and gripper. Using a model-based recognition algorithm the image processing software identifies the gripper and the workpiece and determines their relative position. A deviation vector is calculated and fed into the robot control to guide the gripper.

  3. 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.

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. Fast optical 3D form measurement of aspheres including determination of thickness and wedge and decenter errors

    NASA Astrophysics Data System (ADS)

    Stover, E.; Berger, G.; Wendel, M.; Petter, J.

    2015-10-01

    A method for non-contact 3D form testing of aspheric surfaces including determination of decenter and wedge errors and lens thickness is presented. The principle is based on the absolute measurement capability of multi-wavelength interferometry (MWLI). The approach produces high density 3D shape information and geometric parameters at high accuracy in short measurement times. The system allows inspection of aspheres without restrictions in terms of spherical departures, of segmented and discontinuous optics. The optics can be polished or ground and made of opaque or transparent materials.

  9. 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

  10. 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.

  11. 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.

  12. Nature of heterophase inclusions in high-purity optical fiber materials as studied with 3D laser ultramicroscopy

    NASA Astrophysics Data System (ADS)

    Ketkova, L. A.

    2015-09-01

    3D laser ultramicroscopy (3D LUM) is intended specially for determining the concentration and size distribution of submicron inclusions in the bulk samples of high-purity materials for visible and IR fiber optics. In this work the 3D LUM technique is shown to be able to identify the nature of individual inclusions detected. The measurement of the light scattered by an inclusion at a varied probe beam wavelength and polarization and at a varied scattered light collection angle makes it possible to determine the inclusion refractive index. The 3D LUM possibilities are illustrated by the example of studying the inclusion nature in the As2S3 glass samples prepared by the direct synthesis from elements in a quartz container at elevated temperatures.

  13. 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

  14. 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

  15. 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.

  16. Creation of quantum-degenerate gases of ytterbium in a compact 2D-/3D-magneto-optical trap setup.

    PubMed

    Dörscher, Sören; Thobe, Alexander; Hundt, Bastian; Kochanke, André; Le Targat, Rodolphe; Windpassinger, Patrick; Becker, Christoph; Sengstock, Klaus

    2013-04-01

    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 (1)S0 → (1)P1 transition captures ytterbium directly from a dispenser of atoms and loads a 3D-MOT on the narrow (1)S0 → (3)P1 intercombination transition. Subsequently, atoms are transferred to a crossed optical dipole trap and cooled evaporatively to quantum degeneracy. PMID:23635183

  17. Digital holographic microscopy combined with optical tweezers

    NASA Astrophysics Data System (ADS)

    Cardenas, Nelson; Yu, Lingfeng; Mohanty, Samarendra K.

    2011-02-01

    While optical tweezers have been widely used for the manipulation and organization of microscopic objects in three dimensions, observing the manipulated objects along axial direction has been quite challenging. In order to visualize organization and orientation of objects along axial direction, we report development of a Digital holographic microscopy combined with optical tweezers. Digital holography is achieved by use of a modified Mach-Zehnder interferometer with digital recording of interference pattern of the reference and sample laser beams by use of a single CCD camera. In this method, quantitative phase information is retrieved dynamically with high temporal resolution, only limited by frame rate of the CCD. Digital focusing, phase-unwrapping as well as online analysis and display of the quantitative phase images was performed on a software developed on LabView platform. Since phase changes observed in DHOT is very sensitive to optical thickness of trapped volume, estimation of number of particles trapped in the axial direction as well as orientation of non-spherical objects could be achieved with high precision. Since in diseases such as malaria and diabetics, change in refractive index of red blood cells occurs, this system can be employed to map such disease-specific changes in biological samples upon immobilization with optical tweezers.

  18. 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.

  19. 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.

  20. DMD-based LED-illumination Super-resolution and optical sectioning microscopy

    PubMed Central

    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. PMID:23346373

  1. 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.

  2. Using optically scanned 3D data in the restoration of Michelangelo's David

    NASA Astrophysics Data System (ADS)

    Scopigno, Roberto; Cignoni, Paolo; Callieri, Marco; Ganovelli, Fabio; Impoco, G.; Pingi, P.; Ponchio, F.

    2003-10-01

    Modern 3D scanning technologies allow to reconstruct 3D digital representations of Cultural Heritage artifacts in a semi-automatic way, characterized by very high accuracy and wealth of details. The availability of an accurate digital representation opens several possibilities of utilization to experts (restorers, archivists, museum curators), or to ordinary people (students, museum visitors). 3D scanned data are commonly used for the production of animations, interactive visualizations, or virtual reality applications. A much more exciting opportunity is to use these data in the restoration of Cultural Heritage artworks. The integration between 3D graphic and restoration represents an open research field where many new supporting tools are required; the David restoration project has given several starting points and guidelines to the definition and development of innovative solutions. Digital 3D models can be used in two different but not subsidiary modes: as an instrument for the execution of specific investigations and as a supporting media for the archival and integration of all the restoration-related information, gathered with the different studies and analysis performed on the artwork. In this paper we present some recent work done in the framework of the Michelangelo's David restoration project. A 3D model of the David was reconstructed by the Digital Michelangelo Project, using laser-based 3D scanning technology. We have developed some tools to make those data accessible and useful in the restoration. Preliminary results are reported here together with some directions for further research.

  3. 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

  4. 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.

  5. 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.

  6. All-optical photoacoustic microscopy using a MEMS scanning mirror

    NASA Astrophysics Data System (ADS)

    Chen, Sung-Liang; Xie, Zhixing; Ling, Tao; Wei, Xunbin; Guo, L. Jay; Wang, Xueding

    2013-03-01

    It has been studied that a potential marker to obtain prognostic information about bladder cancer is tumor neoangiogenesis, which can be quantified by morphometric characteristics such as microvascular density. Photoacoustic microscopy (PAM) can render sensitive three-dimensional (3D) mapping of microvasculature, providing promise to evaluate the neoangiogenesis that is closely related to the diagnosis of bladder cancer. To ensure good image quality, it is desired to acquire bladder PAM images from its inside via the urethra, like conventional cystoscope. Previously, we demonstrated all-optical PAM systems using polymer microring resonators to detect photoacoustic signals and galvanometer mirrors for laser scanning. In this work, we build a miniature PAM system using a microelectromechanical systems (MEMS) scanning mirror, demonstrating a prototype of an endoscopic PAM head capable of high imaging quality of the bladder. The system has high resolutions of 17.5 μm in lateral direction and 19 μm in the axial direction at a distance of 5.4 mm. Images of printed grids and the 3D structure of microvasculature in animal bladders ex vivo by the system are demonstrated.

  7. 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

  8. 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).

  9. 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.

  10. 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

  11. 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

  12. 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.

  13. Combined transmission and reflection optical microscopy of ice core sections

    NASA Astrophysics Data System (ADS)

    Binder, Tobias; Weikusat, Ilka; Kerst, Thomas; Eichler, Jan; Svensson, Anders; Bohleber, Pascal; Garbe, Christoph; Kipfstuhl, Sepp

    2013-04-01

    information and microscopy in reflection mode gains information on the surface, an "optimal" matching of both images contains displacements of grain boundary sites. We try to quantify this inaccuracy which can also be interpreted as orientation of the grain boundary surface in 3D. [1] T. Binder et al., 2013, Journal of Microscopy, in review [2] I. Weikusat et al., 2011, Journal of Glaciology, 57, 111-120

  14. Reducing depth induced spherical aberration in 3D widefield fluorescence microscopy by wavefront coding using the SQUBIC phase mask

    NASA Astrophysics Data System (ADS)

    Patwary, Nurmohammed; Doblas, Ana; King, Sharon V.; Preza, Chrysanthe

    2014-03-01

    Imaging thick biological samples introduces spherical aberration (SA) due to refractive index (RI) mismatch between specimen and imaging lens immersion medium. SA increases with the increase of either depth or RI mismatch. Therefore, it is difficult to find a static compensator for SA1. Different wavefront coding methods2,3 have been studied to find an optimal way of static wavefront correction to reduce depth-induced SA. Inspired by a recent design of a radially symmetric squared cubic (SQUBIC) phase mask that was tested for scanning confocal microscopy1 we have modified the pupil using the SQUBIC mask to engineer the point spread function (PSF) of a wide field fluorescence microscope. In this study, simulated images of a thick test object were generated using a wavefront encoded engineered PSF (WFEPSF) and were restored using space-invariant (SI) and depth-variant (DV) expectation maximization (EM) algorithms implemented in the COSMOS software4. Quantitative comparisons between restorations obtained with both the conventional and WFE PSFs are presented. Simulations show that, in the presence of SA, the use of the SIEM algorithm and a single SQUBIC encoded WFE-PSF can yield adequate image restoration. In addition, in the presence of a large amount of SA, it is possible to get adequate results using the DVEM with fewer DV-PSFs than would typically be required for processing images acquired with a clear circular aperture (CCA) PSF. This result implies that modification of a widefield system with the SQUBIC mask renders the system less sensitive to depth-induced SA and suitable for imaging samples at larger optical depths.

  15. Shape measurement by a multi-view methodology based on the remote tracking of a 3D optical scanner

    NASA Astrophysics Data System (ADS)

    Barone, Sandro; Paoli, Alessandro; Viviano Razionale, Armando

    2012-03-01

    Full field optical techniques can be reliably used for 3D measurements of complex shapes by multi-view processes, which require the computation of transformation parameters relating different views into a common reference system. Although, several multi-view approaches have been proposed, the alignment process is still the crucial step of a shape reconstruction. In this paper, a methodology to automatically align 3D views has been developed by integrating a stereo vision system and a full field optical scanner. In particular, the stereo vision system is used to remotely track the optical scanner within a working volume. The tracking system uses stereo images to detect the 3D coordinates of retro-reflective infrared markers rigidly connected to the scanner. Stereo correspondences are established by a robust methodology based on combining the epipolar geometry with an image spatial transformation constraint. The proposed methodology has been validated by experimental tests regarding both the evaluation of the measurement accuracy and the 3D reconstruction of an industrial shape.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. Options in virtual 3D, optical-impression-based planning of dental implants.

    PubMed

    Reich, Sven; Kern, Thomas; Ritter, Lutz

    2014-01-01

    If a 3D radiograph, which in today's dentistry often consists of a CBCT dataset, is available for computerized implant planning, the 3D planning should also consider functional prosthetic aspects. In a conventional workflow, the CBCT is done with a specially produced radiopaque prosthetic setup that makes the desired prosthetic situation visible during virtual implant planning. If an exclusively digital workflow is chosen, intraoral digital impressions are taken. On these digital models, the desired prosthetic suprastructures are designed. The entire datasets are virtually superimposed by a "registration" process on the corresponding structures (teeth) in the CBCTs. Thus, both the osseous and prosthetic structures are visible in one single 3D application and make it possible to consider surgical and prosthetic aspects. After having determined the implant positions on the computer screen, a drilling template is designed digitally. According to this design (CAD), a template is printed or milled in CAM process. This template is the first physically extant product in the entire workflow. The article discusses the options and limitations of this workflow. PMID:25098158

  2. 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

  3. Increasing the filling fraction of ultracold polar KRb molecules in a 3D optical lattice

    NASA Astrophysics Data System (ADS)

    Moses, Steven; Gadway, Bryce; Yan, Bo; Covey, Jacob; Jin, Deborah; Ye, Jun

    2013-05-01

    Gases of ultracold polar molecules with sufficiently low entropy are an ideal experimental scenario to look for signatures of long-range many-body interactions. Having a high filling fraction in a 3D lattice is one way to achieve a low entropy state. In earlier work, we showed that preformed pairs of K and Rb in a 3D lattice (sites that have exactly one K and one Rb) are converted to KRb Feshbach molecules with nearly 100% efficiency. Since the STIRAP transfer from Feshbach molecules to ground-state molecules is 90-100% efficient, loading a 3D lattice with a large fraction of preformed pairs will lead to a large filling fraction of ground-state molecules. Our scheme is to load a Mott insulator of Rb and band insulator of K. After we have loaded a lattice with a high filling fraction, we will investigate effects of dipolar interactions in a many-body system. We acknowledge funding from NIST, NSF, AFOSR-MURI, and the NDSEG Graduate Fellowship.

  4. Sub-cellular quantitative optical diffraction tomography with digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Charrière, Florian; Kühn, Jonas; Colomb, Tristan; Cuche, Etienne; Marquet, Pierre; Depeursinge, Christian

    2007-02-01

    Digital holographic microscopy (DHM) is an interferometric technique, providing quantitative mapping of the phase shift induced by semi-transparent microscopic specimens, such as cells, with sub-wavelength resolution along the optical axis. Thanks to actual PCs and CCDs, DHM provides nowadays cost-effective instruments for real-time measurements at very high acquisition rates, with sub-micron transverse resolution. However, DHM phase images do not reveal the threedimensional (3D) internal distribution of refractive index, but a phase shift resulting from a mean refractive index (RI) integrated over the cellular thickness. Standard optical diffraction tomography (ODT) techniques can be efficiently applied to reveal internal structures and to measure 3D RI spatial distributions, by recording 2D DHM phase data for different sample orientations or illumination beam direction, in order to fill up entirely the Ewald sphere in the Fourier space. The 3D refractive index can then be reconstructed, even in the direct space with backpropagation algorithms or from the Fourier space with inverse Fourier transform. The presented technique opens wide perspectives in 3D cell imaging: the DHM-based micro-tomography furnishes invaluable data on the cell components optical properties, potentially leading to information about organelles intracellular distribution. Results obtained on biological specimens will be presented. Morphometric measurements can be extracted from the tomographic data, by detection based on the refractive index contrast within the 3D reconstructions. Results and perspectives about sub-cellular organelles identification inside the cell will also be exposed.

  5. Improved optical fiber probes for scanning near field optical microscopy

    NASA Astrophysics Data System (ADS)

    Wheaton, Bryan R.

    2004-12-01

    The motivation behind this work stems from a combination of my interest in atomic force microscopy (AFM) and the need to apply AFM to several areas of glass research. AFM was used as the main characterization tool in the study of near-field scanning optical microscopy (NSOM) tip formation, evaluation of phase separation in glasses and copper oxide semiconductor film formation. The use of atomic force microscopy (AFM) to evaluate the evolving tip structure of an optical fiber probe for NSOM was studied. This study demonstrates the feasibility of predicting the final tip cone angle, without taking the etching process to completion. Cone angles reported in this study ranged from 58 to 152 degrees, depending on the fiber type and etch conditions. The ability to vary the probe cone angle, and utilize AFM to evaluate the cone angle that results from a set of etch conditions, are valuable additions to the development of NSOM fiber tips. The chemical and spatial variation of phase separated morphologies in glasses can range from a few angstroms to microns, often requiring very high magnification for detection. Historically phase separated glasses have been characterized by transmission electron microscopy (TEM), a time consuming and costly technique. Atomic force microscopy (AFM) provides an inexpensive alternative to TEM and has proven to be a powerful tool in the evaluation of type, degree and scale of phase separation in glasses down to the nanometer level. AFM was used to show that the thickness and uniformity of the CuO films grown in-situ on the surface of copper containing alkali borosilicate glasses increased with time and temperature, however an upper time limit was reached in which no further thickness increases were realized. Tenorite, cuprite and copper metal films were produced depending on the heat treatment environment. XPS was utilized to confirm that copper oxide film formation during heat treatments of glasses near Tg results from the oxidation of copper

  6. 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

  7. Scaled Heavy-Ball Acceleration of the Richardson-Lucy Algorithm for 3D Microscopy Image Restoration.

    PubMed

    Wang, Hongbin; Miller, Paul C

    2014-02-01

    The Richardson-Lucy algorithm is one of the most important in image deconvolution. However, a drawback is its slow convergence. A significant acceleration was obtained using the technique proposed by Biggs and Andrews (BA), which is implemented in the deconvlucy function of the image processing MATLAB toolbox. The BA method was developed heuristically with no proof of convergence. In this paper, we introduce the heavy-ball (H-B) method for Poisson data optimization and extend it to a scaled H-B method, which includes the BA method as a special case. The method has a proof of the convergence rate of O(K(-2)), where k is the number of iterations. We demonstrate the superior convergence performance, by a speedup factor of five, of the scaled H-B method on both synthetic and real 3D images. PMID:26270922

  8. Automated analysis of 3D morphology of human red blood cells via off-axis digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Moon, Inkyu

    2013-05-01

    In this paper we overview an automated method for the analysis of clinical parameters of human red blood cells (RBCs). The digital holograms of mature RBCs are recorded by CCD camera with off-axis interferometry setup and the quantitative phase images of RBCs are formed by a numerical reconstruction technique. For automated investigation of the 3D morphology and mean corpuscular hemoglobin of RBCs, the unnecessary background in the RBCs phase images are removed by marker-controlled watershed segmentation algorithm. Then, characteristic properties of each RBC such as projected cell surface, average phase, mean corpuscular hemoglobin (MCH) and (MCH) surface density is quantitatively measured. Finally, the equality of covariance matrixes and mean vectors of these features for different kinds of RBCs are experimentally analyzed using statistical test scheme. Results show that these characteristic parameters of RBCs can be used as feature pattern to discriminate between RBC populations that differ in shape and hemoglobin content.

  9. Interaction of coherent confined optical modes in neighboring 3D cylindrical ZnO microcavities

    SciTech Connect

    Gruzintsev, A. N. Volkov, V. T.; Knyazev, M. A.; Yakimov, E. E.

    2006-11-15

    Luminescent properties of pairs of neighboring variously spaced 3D cylindrical ZnO microcavities 1.8 {mu}m in diameter, produced by electron-beam lithography and reactive ion etching, are studied. Narrow luminescence peaks in the ZnO exciton spectral region, related to single-mode lasing, were observed. The energy of exchange interaction of coupled modes of two microcavities was calculated as a function of the intercavity distance. Broadening of the line of stimulated UV luminescence associated with coupled photonic modes of two microcavities was observed as the intercavity distance decreased.

  10. 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.

  11. 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…

  12. Numerical Simulation of Two-grid Ion Optics Using a 3D Code

    NASA Technical Reports Server (NTRS)

    Anderson, John R.; Katz, Ira; Goebel, Dan

    2004-01-01

    A three-dimensional ion optics code has been developed under NASA's Project Prometheus to model two grid ion optics systems. The code computes the flow of positive ions from the discharge chamber through the ion optics and into the beam downstream of the thruster. The rate at which beam ions interact with background neutral gas to form charge exchange ions is also computed. Charge exchange ion trajectories are computed to determine where they strike the ion optics grid surfaces and to determine the extent of sputter erosion they cause. The code has been used to compute predictions of the erosion pattern and wear rate on the NSTAR ion optics system; the code predicts the shape of the eroded pattern but overestimates the initial wear rate by about 50%. An example of use of the code to estimate the NEXIS thruster accelerator grid life is also presented.

  13. 3D-localization microscopy and tracking of FoF1-ATP synthases in living bacteria

    NASA Astrophysics Data System (ADS)

    Renz, Anja; Renz, Marc; Klütsch, Diana; Deckers-Hebestreit, Gabriele; Börsch, Michael

    2015-03-01

    FoF1-ATP synthases are membrane-embedded protein machines that catalyze the synthesis of adenosine triphosphate. Using photoactivation-based localization microscopy (PALM) in TIR-illumination as well as structured illumination microscopy (SIM), we explore the spatial distribution and track single FoF1-ATP synthases in living E. coli cells under physiological conditions at different temperatures. For quantitative diffusion analysis by mean-squared-displacement measurements, the limited size of the observation area in the membrane with its significant membrane curvature has to be considered. Therefore, we applied a 'sliding observation window' approach (M. Renz et al., Proc. SPIE 8225, 2012) and obtained the one-dimensional diffusion coefficient of FoF1-ATP synthase diffusing on the long axis in living E. coli cells.

  14. Do's and don'ts of cryo-electron microscopy: a primer on sample preparation and high quality data collection for macromolecular 3D reconstruction.

    PubMed

    Cabra, Vanessa; Samsó, Montserrat

    2015-01-01

    Cryo-electron microscopy (cryoEM) entails flash-freezing a thin layer of sample on a support, and then visualizing the sample in its frozen hydrated state by transmission electron microscopy (TEM). This can be achieved with very low quantity of protein and in the buffer of choice, without the use of any stain, which is very useful to determine structure-function correlations of macromolecules. When combined with single-particle image processing, the technique has found widespread usefulness for 3D structural determination of purified macromolecules. The protocol presented here explains how to perform cryoEM and examines the causes of most commonly encountered problems for rational troubleshooting; following all these steps should lead to acquisition of high quality cryoEM images. The technique requires access to the electron microscope instrument and to a vitrification device. Knowledge of the 3D reconstruction concepts and software is also needed for computerized image processing. Importantly, high quality results depend on finding the right purification conditions leading to a uniform population of structurally intact macromolecules. The ability of cryoEM to visualize macromolecules combined with the versatility of single particle image processing has proven very successful for structural determination of large proteins and macromolecular machines in their near-native state, identification of their multiple components by 3D difference mapping, and creation of pseudo-atomic structures by docking of x-ray structures. The relentless development of cryoEM instrumentation and image processing techniques for the last 30 years has resulted in the possibility to generate de novo 3D reconstructions at atomic resolution level. PMID:25651412

  15. Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

    PubMed Central

    Cabra, Vanessa; Samsó, Montserrat

    2015-01-01

    Cryo-electron microscopy (cryoEM) entails flash-freezing a thin layer of sample on a support, and then visualizing the sample in its frozen hydrated state by transmission electron microscopy (TEM). This can be achieved with very low quantity of protein and in the buffer of choice, without the use of any stain, which is very useful to determine structure-function correlations of macromolecules. When combined with single-particle image processing, the technique has found widespread usefulness for 3D structural determination of purified macromolecules. The protocol presented here explains how to perform cryoEM and examines the causes of most commonly encountered problems for rational troubleshooting; following all these steps should lead to acquisition of high quality cryoEM images. The technique requires access to the electron microscope instrument and to a vitrification device. Knowledge of the 3D reconstruction concepts and software is also needed for computerized image processing. Importantly, high quality results depend on finding the right purification conditions leading to a uniform population of structurally intact macromolecules. The ability of cryoEM to visualize macromolecules combined with the versatility of single particle image processing has proven very successful for structural determination of large proteins and macromolecular machines in their near-native state, identification of their multiple components by 3D difference mapping, and creation of pseudo-atomic structures by docking of x-ray structures. The relentless development of cryoEM instrumentation and image processing techniques for the last 30 years has resulted in the possibility to generate de novo 3D reconstructions at atomic resolution level. PMID:25651412

  16. Segmentation of Whole Cells and Cell Nuclei From 3-D Optical Microscope Images Using Dynamic Programming

    PubMed Central

    McCullough, Dean P.; Gudla, Prabhakar R.; Harris, Bradley S.; Collins, Jason A.; Meaburn, Karen J.; Nakaya, Masa-Aki; Yamaguchi, Terry P.; Misteli, Tom; Lockett, Stephen J.

    2009-01-01

    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. PMID:18450544

  17. 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

  18. Detecting the Chern number of topological Weyl semimetals in 3D optical lattices

    NASA Astrophysics Data System (ADS)

    Zhang, Dan-Wei; Cao, Shuai

    2016-06-01

    We propose a realistic scheme to directly probe the Chern number of topological Weyl semimetals in optical lattices. The Weyl semimetal states can be realized with ultracold fermionic atoms trapped in three-dimensional optical lattices, and are topologically characterized by k z -dependent Chern number, where k z is the out-of-plane quasimomentum. We demonstrate with numerical simulations that this characteristic topological invariant can be extracted from the shift of the hybrid Wannier center in the optical lattice, based on the particle pumping approach. Through in situ measurement of atomic density, the topological properties of the Weyl semimetal states are then directly revealed.

  19. 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.

  20. Brillouin Optical Microscopy for Corneal Biomechanics

    PubMed Central

    Scarcelli, Giuliano; Pineda, Roberto

    2012-01-01

    Purpose. The mechanical properties of corneal tissue are linked to prevalent ocular diseases and therapeutic procedures. Brillouin microscopy is a novel optical technology that enables three-dimensional mechanical imaging. In this study, the feasibility of this noncontact technique was tested for in situ quantitative assessment of the biomechanical properties of the cornea. Methods. Brillouin light-scattering involves a spectral shift proportional to the longitudinal modulus of elasticity of the tissue. A 532-nm single-frequency laser and a custom-developed ultrahigh-resolution spectrometer were used to measure the Brillouin frequency. Confocal scanning was used to perform Brillouin elasticity imaging of the corneas of whole bovine eyes. The longitudinal modulus of the bovine corneas was compared before and after riboflavin corneal collagen photo-cross-linking. The Brillouin measurements were then compared with conventional stress–strain mechanical test results. Results. High-resolution Brillouin images of the cornea were obtained, revealing a striking depth-dependent variation of the elastic modulus across the cornea. Along the central axis, the Brillouin frequency shift varied gradually from 8.2 GHz in the epithelium to 7.5 GHz near the endothelium. The coefficients of the down slope were measured to be approximately 1.09, 0.32, and 2.94 GHz/mm in the anterior, posterior, and innermost stroma, respectively. On riboflavin collagen cross-linking, marked changes in the axial Brillouin profiles (P < 0.001) were noted before and after cross-linking. Conclusions. Brillouin imaging can assess the biomechanical properties of cornea in situ with high spatial resolution. This novel technique has the potential for use in clinical diagnostics and treatment monitoring. PMID:22159012