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Sample records for 3d optical microscopy

  1. 3D high resolution pure optical photoacoustic microscopy

    NASA Astrophysics Data System (ADS)

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

    2012-02-01

    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 some refinedment of 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 of 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 5μm and an axial resolution of 8μm, 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 properties at depths in unfrozen tissue

  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. Cytology 3D structure formation based on optical microscopy images

    NASA Astrophysics Data System (ADS)

    Pronichev, A. N.; Polyakov, E. V.; Shabalova, I. P.; Djangirova, T. V.; Zaitsev, S. M.

    2017-01-01

    The article the article is devoted to optimization of the parameters of imaging of biological preparations in optical microscopy using a multispectral camera in visible range of electromagnetic radiation. A model for the image forming of virtual preparations was proposed. The optimum number of layers was determined for the object scan in depth and holistic perception of its switching according to the results of the experiment.

  4. 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.

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

    NASA Astrophysics Data System (ADS)

    Grover, Ginni

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

  6. 3D structured illumination microscopy

    NASA Astrophysics Data System (ADS)

    Dougherty, William M.; Goodwin, Paul C.

    2011-03-01

    Three-dimensional structured illumination microscopy achieves double the lateral and axial resolution of wide-field microscopy, using conventional fluorescent dyes, proteins and sample preparation techniques. A three-dimensional interference-fringe pattern excites the fluorescence, filling in the "missing cone" of the wide field optical transfer function, thereby enabling axial (z) discrimination. The pattern acts as a spatial carrier frequency that mixes with the higher spatial frequency components of the image, which usually succumb to the diffraction limit. The fluorescence image encodes the high frequency content as a down-mixed, moiré-like pattern. A series of images is required, wherein the 3D pattern is shifted and rotated, providing down-mixed data for a system of linear equations. Super-resolution is obtained by solving these equations. The speed with which the image series can be obtained can be a problem for the microscopy of living cells. Challenges include pattern-switching speeds, optical efficiency, wavefront quality and fringe contrast, fringe pitch optimization, and polarization issues. We will review some recent developments in 3D-SIM hardware with the goal of super-resolved z-stacks of motile cells.

  7. 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.

  8. Deep Learning Segmentation of Optical Microscopy Images Improves 3D Neuron Reconstruction.

    PubMed

    Li, Rongjian; Zeng, Tao; Peng, Hanchuan; Ji, Shuiwang

    2017-03-08

    Digital reconstruction, or tracing, of 3-dimensional (3D) neuron structure from microscopy images is a critical step toward reversing engineering the wiring and anatomy of a brain. Despite a number of prior attempts, this task remains very challenging, especially when images are contaminated by noises or have discontinued segments of neurite patterns. An approach for addressing such problems is to identify the locations of neuronal voxels using image segmentation methods prior to applying tracing or reconstruction techniques. This preprocessing step is expected to remove noises in the data, thereby leading to improved reconstruction results. In this work, we proposed to use 3D Convolutional neural networks (CNNs) for segmenting the neuronal microscopy images. Specifically, we designed a novel CNN architecture that takes volumetric images as the inputs and their voxel-wise segmentation maps as the outputs. The developed architecture allows us to train and predict using large microscopy images in an end-to-end manner. We evaluated the performance of our model on a variety of challenging 3D microscopy images from different organisms. Results showed that the proposed methods improved the tracing performance significantly when combined with different reconstruction algorithms.

  9. 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

  10. 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.

  11. Pico-projector-based optical sectioning microscopy for 3D chlorophyll fluorescence imaging of mesophyll cells

    NASA Astrophysics Data System (ADS)

    Chen, Szu-Yu; Hsu, Yu John; Yeh, Chia-Hua; Chen, S.-Wei; Chung, Chien-Han

    2015-03-01

    A pico-projector-based optical sectioning microscope (POSM) was constructed using a pico-projector to generate structured illumination patterns. A net rate of 5.8 × 106 pixel/s and sub-micron spatial resolution in three-dimensions (3D) were achieved. Based on the pico-projector’s flexibility in pattern generation, the characteristics of POSM with different modulation periods and at different imaging depths were measured and discussed. With the application of different modulation periods, 3D chlorophyll fluorescence imaging of mesophyll cells was carried out in freshly plucked leaves of four species without sectioning or staining. For each leaf, an average penetration depth of 120 μm was achieved. Increasing the modulation period along with the increment of imaging depth, optical sectioning images can be obtained with a compromise between the axial resolution and signal-to-noise ratio. After ∼30 min imaging on the same area, photodamage was hardly observed. Taking the advantages of high speed and low damages of POSM, the investigation of the dynamic fluorescence responses to temperature changes was performed under three different treatment temperatures. The three embedded blue, green and red light-emitting diode light sources were applied to observe the responses of the leaves with different wavelength excitation.

  12. Measurement of dynamic cell-induced 3D displacement fields in vitro for traction force optical coherence microscopy

    PubMed Central

    Mulligan, Jeffrey A.; Bordeleau, François; Reinhart-King, Cynthia A.; Adie, Steven G.

    2017-01-01

    Traction force microscopy (TFM) is a method used to study the forces exerted by cells as they sense and interact with their environment. Cell forces play a role in processes that take place over a wide range of spatiotemporal scales, and so it is desirable that TFM makes use of imaging modalities that can effectively capture the dynamics associated with these processes. To date, confocal microscopy has been the imaging modality of choice to perform TFM in 3D settings, although multiple factors limit its spatiotemporal coverage. We propose traction force optical coherence microscopy (TF-OCM) as a novel technique that may offer enhanced spatial coverage and temporal sampling compared to current methods used for volumetric TFM studies. Reconstructed volumetric OCM data sets were used to compute time-lapse extracellular matrix deformations resulting from cell forces in 3D culture. These matrix deformations revealed clear differences that can be attributed to the dynamic forces exerted by normal versus contractility-inhibited NIH-3T3 fibroblasts embedded within 3D Matrigel matrices. Our results are the first step toward the realization of 3D TF-OCM, and they highlight the potential use of OCM as a platform for advancing cell mechanics research. PMID:28271010

  13. 3D image reconstruction using optical sectioning in confocal scanning microscopy

    NASA Astrophysics Data System (ADS)

    Seo, Jungwoo; Kang, Dong Kyun; Park, Sunglim; Gweon, Dae gab

    2001-10-01

    Confocal scanning microscopy (CSM) has been used in biological application, materials science, semiconductor quality measurement and other non-destructive microscopic application. Small spot of light illuminates a sample, and a small detector that is ideally a point detector collects the reflected or transmitted light having the information of specimen. An image distribution can be reconstructed by a correlation analysis of spots with the high bandwidth. The mechanism for two-dimensional beam scanning and optical sectioning has an important role in CSM as the three-dimensional profiler. The parasitic motion of focus on the detector gives rise to the fatal distortion of an image profile named the extinction effect while using acousto-optical (AO) deflector. The intensity profile for the open loop scanning should be matched with its response for the standard. The non-linearity can be minimized with the optical sectioning or the optical probe of the closed loop control. This paper shows the mathematical expression of the light such as the extinction curve in the optical fields of system using AO deflector, the axial/lateral response experimentally when the error sources change, and the methods of optical sectioning. We propose the progressive methods for the high quality image as the following. At first, for having the corrected image, small spot and long scan range, this paper shows that the optimal design having the multi-objects can be used by choosing the unitary lens device in CSM. At second, in order to compensate for the intensity cancellation at the end profile that may be the cause of waviness for the optical image, this paper shows that it is efficient to schedule the frequency of scan. According to characteristics of the extinction curve and axial/lateral response having the error property, we can define the frequency and sensitivity of as their robustness. Finally, the axial response gives an important motive for the optical section, and the limit of

  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. Spatial light modulator phase mask implementation of wavefront encoded 3D computational-optical microscopy.

    PubMed

    King, Sharon V; Doblas, Ana; Patwary, Nurmohammed; Saavedra, Genaro; Martínez-Corral, Manuel; Preza, Chrysanthe

    2015-10-10

    Spatial light modulator (SLM) implementation of wavefront encoding enables various types of engineered point-spread functions (PSFs), including the generalized-cubic and squared-cubic phase mask wavefront encoded (WFE) PSFs, shown to reduce the impact of sample-induced spherical aberration in fluorescence microscopy. This investigation validates dynamic experimental parameter variation of these WFE-PSFs. We find that particular design parameter bounds exist, within which the divergence of computed and experimental WFE-PSFs is of the same order of magnitude as that of computed and experimental conventional PSFs, such that model-based approaches for solving the inverse imaging problem can be applied to a wide range of SLM-WFE systems. Interferometric measurements were obtained to evaluate the SLM implementation of the numeric mask. Agreement between experiment and theory in terms of a wrapped phase, 0-2π, validates the phase mask implementation and allows characterization of the SLM response. These measurements substantiate experimental practice of computational-optical microscope imaging with an SLM-engineered PSF.

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

    PubMed Central

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

    2015-01-01

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

  18. 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.

  19. 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

  20. Calibration-free absolute quantification of optical absorption coefficients using acoustic spectra in 3D photoacoustic microscopy of biological tissue.

    PubMed

    Guo, Zijian; Hu, Song; Wang, Lihong V

    2010-06-15

    Optical absorption is closely associated with many physiological important parameters, such as the concentration and oxygen saturation of hemoglobin, and it can be used to quantify the concentrations of nonfluorescent molecules. We propose a method to use acoustic spectra of photoacoustic signals to quantify the absolute optical absorption. This method is self-calibrating and thus insensitive to variations in the optical fluence. Factors such as system bandwidth and acoustic attenuation can affect the quantification but can be canceled by dividing the acoustic spectra measured at two optical wavelengths. Using optical-resolution photoacoustic microscopy, we quantified the absolute optical absorption of black ink samples with various concentrations. We also quantified both the concentration and oxygen saturation of hemoglobin in a live mouse in absolute units.

  1. Atomic resolution 3D electron diffraction microscopy

    SciTech Connect

    Miao, Jianwei; Ohsuna, Tetsu; Terasaki, Osamu; O'Keefe, Michael A.

    2002-03-01

    Electron lens aberration is the major barrier limiting the resolution of electron microscopy. Here we describe a novel form of electron microscopy to overcome electron lens aberration. By combining coherent electron diffraction with the oversampling phasing method, we show that the 3D structure of a 2 x 2 x 2 unit cell nano-crystal (framework of LTA [Al12Si12O48]8) can be ab initio determined at the resolution of 1 Angstrom from a series of simulated noisy diffraction pattern projections with rotation angles ranging from -70 degrees to +70 degrees in 5 degrees increments along a single rotation axis. This form of microscopy (which we call 3D electron diffraction microscopy) does not require any reference waves, and can image the 3D structure of nanocrystals, as well as non-crystalline biological and materials science samples, with the resolution limited only by the quality of sample diffraction.

  2. 3D differential phase contrast microscopy

    NASA Astrophysics Data System (ADS)

    Chen, Michael; Tian, Lei; Waller, Laura

    2016-03-01

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

  3. 3D Viscoelastic traction force microscopy.

    PubMed

    Toyjanova, Jennet; Hannen, Erin; Bar-Kochba, Eyal; Darling, Eric M; Henann, David L; Franck, Christian

    2014-10-28

    Native cell-material interactions occur on materials differing in their structural composition, chemistry, and physical compliance. While the last two decades have shown the importance of traction forces during cell-material interactions, they have been almost exclusively presented on purely elastic in vitro materials. Yet, most bodily tissue materials exhibit some level of viscoelasticity, which could play an important role in how cells sense and transduce tractions. To expand the realm of cell traction measurements and to encompass all materials from elastic to viscoelastic, this paper presents a general, and comprehensive approach for quantifying 3D cell tractions in viscoelastic materials. This methodology includes the experimental characterization of the time-dependent material properties for any viscoelastic material with the subsequent mathematical implementation of the determined material model into a 3D traction force microscopy (3D TFM) framework. Utilizing this new 3D viscoelastic TFM (3D VTFM) approach, we quantify the influence of viscosity on the overall material traction calculations and quantify the error associated with omitting time-dependent material effects, as is the case for all other TFM formulations. We anticipate that the 3D VTFM technique will open up new avenues of cell-material investigations on even more physiologically relevant time-dependent materials including collagen and fibrin gels.

  4. 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.

  5. Stroboscobic near-field scanning optical microscopy for 3D mapping of mode profiles of plasmonic nanostructures (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Dana, Aykutlu; Ozgur, Erol; Torunoglu, Gamze

    2016-09-01

    We present a dynamic approach to scanning near field optical microscopy that extends the measurement technique to the third dimension, by strobing the illumination in sync with the cantilever oscillation. Nitrogen vacancy (NV) centers in nanodiamonds placed on cantilever tips are used as stable emitters for emission enhancement. Local field enhancement and modulation of optical density states are mapped in three dimensions based on fluorescence intensity and spectrum changes as the tip is scanned over plasmonic nanostructures. The excitation of NV centers is done using a total internal reflection setup. Using a digital phase locked loop to pulse the excitation in various tip sample separations, 2D slices of fluorescence enhancement can be recorded. Alternatively, a conventional SNOM tip can be used to selectively couple wideband excitation to the collection path, with subdiffraction resolution of 60 nm in x and y and 10 nm in z directions. The approach solves the problem of tip-sample separation stabilization over extended periods of measurement time, required to collect data resolved in emission wavelength and three spatial dimensions. The method can provide a unique way of accessing the three dimensional field and mode profiles of nanophotonics structures.

  6. Polarized 3D Raman and nanoscale near-field optical microscopy of optically inscribed surface relief gratings: chromophore orientation in azo-doped polymer films.

    PubMed

    Di Florio, Giuseppe; Bründermann, Erik; Yadavalli, Nataraja Sekhar; Santer, Svetlana; Havenith, Martina

    2014-03-14

    We have used polarized confocal Raman microspectroscopy and scanning near-field optical microscopy with a resolution of 60 nm to characterize photoinscribed grating structures of azobenzene doped polymer films on a glass support. Polarized Raman microscopy allowed determining the reorientation of the chromophores as a function of the grating phase and penetration depth of the inscribing laser in three dimensions. We found periodic patterns, which are not restricted to the surface alone, but appear also well below the surface in the bulk of the material. Near-field optical microscopy with nanoscale resolution revealed lateral two-dimensional optical contrast, which is not observable by atomic force and Raman microscopy.

  7. Simple buffers for 3D STORM microscopy.

    PubMed

    Olivier, Nicolas; Keller, Debora; Rajan, Vinoth Sundar; Gönczy, Pierre; Manley, Suliana

    2013-06-01

    3D STORM is one of the leading methods for super-resolution imaging, with resolution down to 10 nm in the lateral direction, and 30-50 nm in the axial direction. However, there is one important requirement to perform this type of imaging: making dye molecules blink. This usually relies on the utilization of complex buffers, containing different chemicals and sensitive enzymatic systems, limiting the reproducibility of the method. We report here that the commercial mounting medium Vectashield can be used for STORM of Alexa-647, and yields images comparable or superior to those obtained with more complex buffers, especially for 3D imaging. We expect that this advance will promote the versatile utilization of 3D STORM by removing one of its entry barriers, as well as provide a more reproducible way to compare optical setups and data processing algorithms.

  8. Axial Plane Optical Microscopy

    PubMed Central

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

    2014-01-01

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

  9. Three-dimensional scanning near field optical microscopy (3D-SNOM) imaging of random arrays of copper nanoparticles: implications for plasmonic solar cell enhancement.

    PubMed

    Ezugwu, Sabastine; Ye, Hanyang; Fanchini, Giovanni

    2015-01-07

    In order to investigate the suitability of random arrays of nanoparticles for plasmonic enhancement in the visible-near infrared range, we introduced three-dimensional scanning near-field optical microscopy (3D-SNOM) imaging as a useful technique to probe the intensity of near-field radiation scattered by random systems of nanoparticles at heights up to several hundred nm from their surface. We demonstrated our technique using random arrays of copper nanoparticles (Cu-NPs) at different particle diameter and concentration. Bright regions in the 3D-SNOM images, corresponding to constructive interference of forward-scattered plasmonic waves, were obtained at heights Δz ≥ 220 nm from the surface for random arrays of Cu-NPs of ∼ 60-100 nm in diameter. These heights are too large to use Cu-NPs in contact of the active layer for light harvesting in thin organic solar cells, which are typically no thicker than 200 nm. Using a 200 nm transparent spacer between the system of Cu-NPs and the solar cell active layer, we demonstrate that forward-scattered light can be conveyed in 200 nm thin film solar cells. This architecture increases the solar cell photoconversion efficiency by a factor of 3. Our 3D-SNOM technique is general enough to be suitable for a large number of other applications in nanoplasmonics.

  10. Three-dimensional scanning near field optical microscopy (3D-SNOM) imaging of random arrays of copper nanoparticles: implications for plasmonic solar cell enhancement

    NASA Astrophysics Data System (ADS)

    Ezugwu, Sabastine; Ye, Hanyang; Fanchini, Giovanni

    2014-11-01

    In order to investigate the suitability of random arrays of nanoparticles for plasmonic enhancement in the visible-near infrared range, we introduced three-dimensional scanning near-field optical microscopy (3D-SNOM) imaging as a useful technique to probe the intensity of near-field radiation scattered by random systems of nanoparticles at heights up to several hundred nm from their surface. We demonstrated our technique using random arrays of copper nanoparticles (Cu-NPs) at different particle diameter and concentration. Bright regions in the 3D-SNOM images, corresponding to constructive interference of forward-scattered plasmonic waves, were obtained at heights Δz >= 220 nm from the surface for random arrays of Cu-NPs of ~60-100 nm in diameter. These heights are too large to use Cu-NPs in contact of the active layer for light harvesting in thin organic solar cells, which are typically no thicker than 200 nm. Using a 200 nm transparent spacer between the system of Cu-NPs and the solar cell active layer, we demonstrate that forward-scattered light can be conveyed in 200 nm thin film solar cells. This architecture increases the solar cell photoconversion efficiency by a factor of 3. Our 3D-SNOM technique is general enough to be suitable for a large number of other applications in nanoplasmonics.In order to investigate the suitability of random arrays of nanoparticles for plasmonic enhancement in the visible-near infrared range, we introduced three-dimensional scanning near-field optical microscopy (3D-SNOM) imaging as a useful technique to probe the intensity of near-field radiation scattered by random systems of nanoparticles at heights up to several hundred nm from their surface. We demonstrated our technique using random arrays of copper nanoparticles (Cu-NPs) at different particle diameter and concentration. Bright regions in the 3D-SNOM images, corresponding to constructive interference of forward-scattered plasmonic waves, were obtained at heights Δz >= 220

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

    PubMed

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

    2002-04-01

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

  12. Resolution in 3D in multifocal plane microscopy

    NASA Astrophysics Data System (ADS)

    Chao, Jerry; Ram, Sripad; Abraham, Anish V.; Ward, E. Sally; Ober, Raimund J.

    2008-02-01

    Using single molecule microscopy, biological interactions can be imaged and studied at the level of individual biomolecules. When characterizing an imaged biological interaction, the distance separating the two participating biomolecules can provide valuable information. Therefore, the resolvability of an imaging setup is of practical significance in the analysis of the acquired image data. Importantly, the resolvability of the imaging setup needs evaluation in the 3D context, since in general biomolecules reside in 3D space within the cellular environment. We recently introduced an information-theoretic 2D resolution measure which shows that the resolution limit due to Rayleigh's criterion can be overcome. This new result predicts that the resolution of optical microscopes is not limited, but rather can be improved with increased photon counts detected from the single molecules. The 2D result was subsequently extended to the 3D context, and the proposed information-theoretic 3D resolution measure can readily be used to determine the resolvability of a conventional single focal plane imaging setup. Here, we consider the 3D resolution measure for a multifocal plane microscope setup, an imaging system which allows the concurrent imaging of multiple focal planes within a specimen. The technique is useful in applications such as the tracking of subcellular objects in 3D. By comparing their 3D resolution measures, we find a two-plane setup to outperform a comparable conventional single-plane setup in resolvability over a range of axial locations for the single molecule pair. Moreover, we investigate and compare the impact of noise on the resolvability of the two setups.

  13. 3D differential phase contrast microscopy

    PubMed Central

    Chen, Michael; Tian, Lei; Waller, Laura

    2016-01-01

    We demonstrate 3D phase and absorption recovery from partially coherent intensity images captured with a programmable LED array source. Images are captured through-focus with four different illumination patterns. Using first Born and weak object approximations (WOA), a linear 3D differential phase contrast (DPC) model is derived. The partially coherent transfer functions relate the sample’s complex refractive index distribution to intensity measurements at varying defocus. Volumetric reconstruction is achieved by a global FFT-based method, without an intermediate 2D phase retrieval step. Because the illumination is spatially partially coherent, the transverse resolution of the reconstructed field achieves twice the NA of coherent systems and improved axial resolution. PMID:27867705

  14. Frames-Based Denoising in 3D Confocal Microscopy Imaging.

    PubMed

    Konstantinidis, Ioannis; Santamaria-Pang, Alberto; Kakadiaris, Ioannis

    2005-01-01

    In this paper, we propose a novel denoising method for 3D confocal microscopy data based on robust edge detection. Our approach relies on the construction of a non-separable frame system in 3D that incorporates the Sobel operator in dual spatial directions. This multidirectional set of digital filters is capable of robustly detecting edge information by ensemble thresholding of the filtered data. We demonstrate the application of our method to both synthetic and real confocal microscopy data by comparing it to denoising methods based on separable 3D wavelets and 3D median filtering, and report very encouraging results.

  15. Motility imaging via optical coherence phase microscopy enables label-free monitoring of tissue growth and viability in 3D tissue-engineering scaffolds.

    PubMed

    Holmes, Christina; Tabrizian, Maryam; Bagnaninchi, Pierre O

    2015-05-01

    As the field of tissue engineering continues to progress, there is a deep need for non-invasive, label-free imaging technologies that can monitor tissue growth and health within thick three-dimensional (3D) constructs. Amongst the many imaging modalities under investigation, optical coherence tomography (OCT) has emerged as a promising tool, enabling non-destructive in situ characterization of scaffolds and engineered tissues. However, the lack of optical contrast between cells and scaffold materials using this technique remains a challenge. In this communication, we show that mapping the optical phase fluctuations resulting from cellular viability and motility allows for the distinction of live cells from their surrounding scaffold environment. Motility imaging was performed via a common-path optical coherence phase microscope (OCPM), an OCT modality that has been shown to be sensitive to nanometer-level fluctuations. More specifically, we examined the development of human adipose-derived stem cells and/or murine pre-osteoblasts within two distinct scaffold systems, commercially available alginate sponges and custom-microfabricated poly(d, l-lactic-co-glycolic acid) fibrous scaffolds. Cellular motility is demonstrated as an endogenous source of contrast for OCPM, enabling real-time, label-free monitoring of 3D engineered tissue development.

  16. 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.

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

    PubMed Central

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

    2012-01-01

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

  18. 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

  19. 3D imaging of neutron tracks using confocal microscopy

    NASA Astrophysics Data System (ADS)

    Gillmore, Gavin; Wertheim, David; Flowers, Alan

    2016-04-01

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

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

    SciTech Connect

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

    2013-10-01

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

  1. Optical microcavity scanning 3D tomography.

    PubMed

    Di Donato, Andrea; Criante, Luigino; LoTurco, Sara; Farina, Marco

    2014-10-01

    A scanning optical microcavity is exploited to achieve lens-free 3D tomography of microfluidic channels. The microcavity, powered by a low-coherence source, is realized by approaching a cleaved fiber to few tens of micrometers over the sample. The interference of scattered waves inside the cavity shapes the transverse field distribution by focusing the beam and overcoming the diffraction limit due to the optical-fiber numerical aperture. The focusing effect is also preserved in the inner layers of the sample, allowing optical 3D tomography. Analysis of microfluidic channels was demonstrated through this noninvasive technique. Although the experimental setup recalls the well-known fiber-optic Fourier-domain common-path optical coherence tomography, the proposed method has intrinsic characteristics that distinguish it from the former one.

  2. 3D Image Analysis of Geomaterials using Confocal Microscopy

    NASA Astrophysics Data System (ADS)

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

    2009-05-01

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

  3. 3D integral imaging with optical processing

    NASA Astrophysics Data System (ADS)

    Martínez-Corral, Manuel; Martínez-Cuenca, Raúl; Saavedra, Genaro; Javidi, Bahram

    2008-04-01

    Integral imaging (InI) systems are imaging devices that provide auto-stereoscopic images of 3D intensity objects. Since the birth of this new technology, InI systems have faced satisfactorily many of their initial drawbacks. Basically, two kind of procedures have been used: digital and optical procedures. The "3D Imaging and Display Group" at the University of Valencia, with the essential collaboration of Prof. Javidi, has centered its efforts in the 3D InI with optical processing. Among other achievements, our Group has proposed the annular amplitude modulation for enlargement of the depth of field, dynamic focusing for reduction of the facet-braiding effect, or the TRES and MATRES devices to enlarge the viewing angle.

  4. 3D nanopillar optical antenna photodetectors.

    PubMed

    Senanayake, Pradeep; Hung, Chung-Hong; Shapiro, Joshua; Scofield, Adam; Lin, Andrew; Williams, Benjamin S; Huffaker, Diana L

    2012-11-05

    We demonstrate 3D surface plasmon photoresponse in nanopillar arrays resulting in enhanced responsivity due to both Localized Surface Plasmon Resonances (LSPRs) and Surface Plasmon Polariton Bloch Waves (SPP-BWs). The LSPRs are excited due to a partial gold shell coating the nanopillar which acts as a 3D Nanopillar Optical Antenna (NOA) in focusing light into the nanopillar. Angular photoresponse measurements show that SPP-BWs can be spectrally coincident with LSPRs to result in a x2 enhancement in responsivity at 1180 nm. Full-wave Finite Difference Time Domain (FDTD) simulations substantiate both the spatial and spectral coupling of the SPP-BW / LSPR for enhanced absorption and the nature of the LSPR. Geometrical control of the 3D NOA and the self-aligned metal hole lattice allows the hybridization of both localized and propagating surface plasmon modes for enhanced absorption. Hybridized plasmonic modes opens up new avenues in optical antenna design in nanoscale photodetectors.

  5. 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.

  6. 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

  7. 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.

  8. Fluorescence in situ hybridization applications for super-resolution 3D structured illumination microscopy.

    PubMed

    Markaki, Yolanda; Smeets, Daniel; Cremer, Marion; Schermelleh, Lothar

    2013-01-01

    Fluorescence in situ hybridization on three-dimensionally preserved cells (3D-FISH) is an efficient tool to analyze the subcellular localization and spatial arrangement of targeted DNA sequences and RNA transcripts at the single cell level. 3D reconstructions from serial optical sections obtained by confocal laser scanning microscopy (CLSM) have long been considered the gold standard for 3D-FISH analyses. Recent super-resolution techniques circumvent the diffraction-limit of optical resolution and have defined a new state-of-the-art in bioimaging. Three-dimensional structured illumination microscopy (3D-SIM) represents one of these technologies. Notably, 3D-SIM renders an eightfold improved volumetric resolution over conventional imaging, and allows the simultaneous visualization of differently labeled target structures. These features make this approach highly attractive for the analysis of spatial relations and substructures of nuclear targets that escape detection by conventional light microscopy. Here, we focus on the application of 3D-SIM for the visualization of subnuclear 3D-FISH preparations. In comparison with conventional fluorescence microscopy, the quality of 3D-SIM data is dependent to a much greater extent on the optimal sample preparation, labeling and acquisition conditions. We describe typical problems encountered with super-resolution imaging of in situ hybridizations in mammalian tissue culture cells and provide optimized DNA-/(RNA)-FISH protocols including combinations with immunofluorescence staining (Immuno-FISH) and DNA replication labeling using click chemistry.

  9. Hybrid wide-field and scanning microscopy for high-speed 3D imaging.

    PubMed

    Duan, Yubo; Chen, Nanguang

    2015-11-15

    Wide-field optical microscopy is efficient and robust in biological imaging, but it lacks depth sectioning. In contrast, scanning microscopic techniques, such as confocal microscopy and multiphoton microscopy, have been successfully used for three-dimensional (3D) imaging with optical sectioning capability. However, these microscopic techniques are not very suitable for dynamic real-time imaging because they usually take a long time for temporal and spatial scanning. Here, a hybrid imaging technique combining wide-field microscopy and scanning microscopy is proposed to accelerate the image acquisition process while maintaining the 3D optical sectioning capability. The performance was demonstrated by proof-of-concept imaging experiments with fluorescent beads and zebrafish liver.

  10. 3D super-resolution microscopy of bacterial division machinery

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

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

  11. 3D super-resolution imaging by localization microscopy.

    PubMed

    Magenau, Astrid; Gaus, Katharina

    2015-01-01

    Fluorescence microscopy is an important tool in all fields of biology to visualize structures and monitor dynamic processes and distributions. Contrary to conventional microscopy techniques such as confocal microscopy, which are limited by their spatial resolution, super-resolution techniques such as photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM) have made it possible to observe and quantify structure and processes on the single molecule level. Here, we describe a method to image and quantify the molecular distribution of membrane-associated proteins in two and three dimensions with nanometer resolution.

  12. 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

  13. Quantitative 3D imaging of whole, unstained cells by using X-ray diffraction microscopy.

    PubMed

    Jiang, Huaidong; Song, Changyong; Chen, Chien-Chun; Xu, Rui; Raines, Kevin S; Fahimian, Benjamin P; Lu, Chien-Hung; Lee, Ting-Kuo; Nakashima, Akio; Urano, Jun; Ishikawa, Tetsuya; Tamanoi, Fuyuhiko; Miao, Jianwei

    2010-06-22

    Microscopy has greatly advanced our understanding of biology. Although significant progress has recently been made in optical microscopy to break the diffraction-limit barrier, reliance of such techniques on fluorescent labeling technologies prohibits quantitative 3D imaging of the entire contents of cells. Cryoelectron microscopy can image pleomorphic structures at a resolution of 3-5 nm, but is only applicable to thin or sectioned specimens. Here, we report quantitative 3D imaging of a whole, unstained cell at a resolution of 50-60 nm by X-ray diffraction microscopy. We identified the 3D morphology and structure of cellular organelles including cell wall, vacuole, endoplasmic reticulum, mitochondria, granules, nucleus, and nucleolus inside a yeast spore cell. Furthermore, we observed a 3D structure protruding from the reconstructed yeast spore, suggesting the spore germination process. Using cryogenic technologies, a 3D resolution of 5-10 nm should be achievable by X-ray diffraction microscopy. This work hence paves a way for quantitative 3D imaging of a wide range of biological specimens at nanometer-scale resolutions that are too thick for electron microscopy.

  14. Quantitative 3D structured illumination microscopy of nuclear structures.

    PubMed

    Kraus, Felix; Miron, Ezequiel; Demmerle, Justin; Chitiashvili, Tsotne; Budco, Alexei; Alle, Quentin; Matsuda, Atsushi; Leonhardt, Heinrich; Schermelleh, Lothar; Markaki, Yolanda

    2017-05-01

    3D structured illumination microscopy (3D-SIM) is the super-resolution technique of choice for multicolor volumetric imaging. Here we provide a validated sample preparation protocol for labeling nuclei of cultured mammalian cells, image acquisition and registration practices, and downstream image analysis of nuclear structures and epigenetic marks. Using immunostaining and replication labeling combined with image segmentation, centroid mapping and nearest-neighbor analyses in open-source environments, 3D maps of nuclear structures are analyzed in individual cells and normalized to fluorescence standards on the nanometer scale. This protocol fills an unmet need for the application of 3D-SIM to the technically challenging nuclear environment, and subsequent quantitative analysis of 3D nuclear structures and epigenetic modifications. In addition, it establishes practical guidelines and open-source solutions using ImageJ/Fiji and the TANGO plugin for high-quality and routinely comparable data generation in immunostaining experiments that apply across model systems. From sample preparation through image analysis, the protocol can be executed within one week.

  15. 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.

  16. Multi-resolution optical 3D sensor

    NASA Astrophysics Data System (ADS)

    Kühmstedt, Peter; Heinze, Matthias; Schmidt, Ingo; Breitbarth, Martin; Notni, Gunther

    2007-06-01

    A new multi resolution self calibrating optical 3D measurement system using fringe projection technique named "kolibri FLEX multi" will be presented. It can be utilised to acquire the all around shape of small to medium objects, simultaneously. The basic measurement principle is the phasogrammetric approach /1,2,3/ in combination with the method of virtual landmarks for the merging of the 3D single views. The system consists in minimum of two fringe projection sensors. The sensors are mounted on a rotation stage illuminating the object from different directions. The measurement fields of the sensors can be chosen different, here as an example 40mm and 180mm in diameter. In the measurement the object can be scanned at the same time with these two resolutions. Using the method of virtual landmarks both point clouds are calculated within the same world coordinate system resulting in a common 3D-point cloud. The final point cloud includes the overview of the object with low point density (wide field) and a region with high point density (focussed view) at the same time. The advantage of the new method is the possibility to measure with different resolutions at the same object region without any mechanical changes in the system or data post processing. Typical parameters of the system are: the measurement time is 2min for 12 images and the measurement accuracy is below 3μm up to 10 μm. The flexibility makes the measurement system useful for a wide range of applications such as quality control, rapid prototyping, design and CAD/CAM which will be shown in the paper.

  17. Unsupervised noise removal algorithms for 3-D confocal fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Roysam, Badrinath; Bhattacharjya, Anoop K.; Srinivas, Chukka; Szarowski, Donald H.; Turner, James N.

    1992-06-01

    Fast algorithms are presented for effective removal of the noise artifact in 3-D confocal fluorescence microscopy images of extended spatial objects such as neurons. The algorithms are unsupervised in the sense that they automatically estimate and adapt to the spatially and temporally varying noise level in the microscopy data. An important feature of the algorithms is the fact that a 3-D segmentation of the field emerges jointly with the intensity estimate. The role of the segmentation is to limit any smoothing to the interiors of regions and hence avoid the blurring that is associated with conventional noise removal algorithms. Fast computation is achieved by parallel computation methods, rather than by algorithmic or modelling compromises. The noise-removal proceeds iteratively, starting from a set of approximate user- supplied, or default initial guesses of the underlying random process parameters. An expectation maximization algorithm is used to obtain a more precise characterization of these parameters, that are then input to a hierarchical estimation algorithm. This algorithm computes a joint solution of the related problems corresponding to intensity estimation, segmentation, and boundary-surface estimation subject to a combination of stochastic priors and syntactic pattern constraints. Three-dimensional stereoscopic renderings of processed 3-D images of murine hippocampal neurons are presented to demonstrate the effectiveness of the method. The processed images exhibit increased contrast and significant smoothing and reduction of the background intensity while avoiding any blurring of the neuronal structures.

  18. Optical 3D surface digitizing in forensic medicine: 3D documentation of skin and bone injuries.

    PubMed

    Thali, Michael J; Braun, Marcel; Dirnhofer, Richard

    2003-11-26

    Photography process reduces a three-dimensional (3D) wound to a two-dimensional level. If there is a need for a high-resolution 3D dataset of an object, it needs to be three-dimensionally scanned. No-contact optical 3D digitizing surface scanners can be used as a powerful tool for wound and injury-causing instrument analysis in trauma cases. The 3D skin wound and a bone injury documentation using the optical scanner Advanced TOpometric Sensor (ATOS II, GOM International, Switzerland) will be demonstrated using two illustrative cases. Using this 3D optical digitizing method the wounds (the virtual 3D computer model of the skin and the bone injuries) and the virtual 3D model of the injury-causing tool are graphically documented in 3D in real-life size and shape and can be rotated in the CAD program on the computer screen. In addition, the virtual 3D models of the bone injuries and tool can now be compared in a 3D CAD program against one another in virtual space, to see if there are matching areas. Further steps in forensic medicine will be a full 3D surface documentation of the human body and all the forensic relevant injuries using optical 3D scanners.

  19. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

    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 3D 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.

  1. Phase mask optimization for 3D parallax EDF microscopy

    NASA Astrophysics Data System (ADS)

    Beckers, Ingeborg E.; Gierlack, Michael; Höppel, Robert; Landskron, Jürgen

    2014-03-01

    Extended depth-of-field (EDF) microscopy is a well-investigated and very simple method to obtain projection images with an extended depth of focus. Despite its advantages of being a real-time method applicable to any microscopic mode with high lateral resolution that can be simply realized by extending a commercial microscope, the lack of z-correlation is still a problem. In this work we present a combined technique of EDF and stereomicroscopy. By cross-correlation depth information is obtained. Finally, 3D images are reconstructed for best phase masks and simulation results are evaluated experimentally.

  2. Brain morphology imaging by 3D microscopy and fluorescent Nissl staining.

    PubMed

    Lazutkin, A A; Komissarova, N V; Toptunov, D M; Anokhin, K V

    2013-07-01

    Modern optical methods (multiphoton and light-sheet fluorescent microscopy) allow 3D imaging of large specimens of the brain with cell resolution. It is therefore essential to refer the resultant 3D pictures of expression of transgene, protein, and other markers in the brain to the corresponding structures in the atlas. This implies counterstaining of specimens with morphological dyes. However, there are no methods for contrasting large samples of the brain without their preliminary slicing. We have developed a method for fluorescent Nissl staining of whole brain samples. 3D reconstructions of specimens of the hippocampus, olfactory bulbs, and cortex were created. The method can be used for morphological control and evaluation of the effects of various factors on the brain using 3D microscopy technique.

  3. 3D scanning Hall probe microscopy with 700 nm resolution

    NASA Astrophysics Data System (ADS)

    Dede, M.; Akram, R.; Oral, A.

    2016-10-01

    In this report, we present a three dimensional (3D) imaging of magnetic field vector B → (x,y,z) emanating from the magnetic material surfaces using a scanning Hall probe microscopy (3D-SHPM) down to a 700 nm spatial resolution. The Hall probe is used to measure Bz(x,y) on the specimen surface at different heights with the step size of Δz = 250 nm, as we move away from the surface in z direction, until the field decays to zero. These set of images are then used to get ∂Bz(x,y)/∂x and ∂Bz(x,y)/∂y at different z by numerical differentiation. Using the Maxwell's equations in the source free region, Bx(x,y) and By(x,y) can be calculated by integrating ∂Bz(x,y)/∂x and ∂Bz(x,y)/∂y in the z direction. Alternatively, the gradients can also be measured in the Hall gradiometer configuration directly. The operation of the 3D-SHPM is demonstrated by imaging Bx(x,y), By(x,y) and Bz(x,y) on a hard disk specimen at a 700 nm resolution, using both of these methods at 77 K. The system is capable of operating from 300 K down to 4 K range.

  4. Optical scanning holographic microscopy

    NASA Astrophysics Data System (ADS)

    Poon, Ting-Chung; Doh, Kyu B.; Schilling, Bradley W.; Wu, Ming H.; Shinoda, Kazunori K.; Suzuki, Yoshiji

    1995-03-01

    We first review a newly developed 3D imaging technique called optical scanning holography (OSH), and discuss recording and reconstruction of a point object using the principle of OSH. We then derive 3D holographic magnification, using three points configured as a 3D object. Finally, we demonstrated 3D imaging capability of OSH by holographically recording two planar objects at different depths and reconstructing the hologram digitally.

  5. An Automated Pipeline for Dendrite Spine Detection and Tracking of 3D Optical Microscopy Neuron Images of In Vivo Mouse Models

    PubMed Central

    Fan, Jing; Zhou, Xiaobo; Dy, Jennifer G.; Zhang, Yong; Wong, Stephen T. C.

    2009-01-01

    The variations in dendritic branch morphology and spine density provide insightful information about the brain function and possible treatment to neurodegenerative disease, for example investigating structural plasticity during the course of Alzheimer's disease. Most automated image processing methods aiming at analyzing these problems are developed for in vitro data. However, in vivo neuron images provide real time information and direct observation of the dynamics of a disease process in a live animal model. This paper presents an automated approach for detecting spines and tracking spine evolution over time with in vivo image data in an animal model of Alzheimer's disease. We propose an automated pipeline starting with curvilinear structure detection to determine the medial axis of the dendritic backbone and spines connected to the backbone. We, then, propose the adaptive local binary fitting (aLBF) energy level set model to accurately locate the boundary of dendritic structures using the central line of curvilinear structure as initialization. To track the growth or loss of spines, we present a maximum likelihood based technique to find the graph homomorphism between two image graph structures at different time points. We employ dynamic programming to search for the optimum solution. The pipeline enables us to extract dynamically changing information from real time in vivo data. We validate our proposed approach by comparing with manual results generated by neurologists. In addition, we discuss the performance of 3D based segmentation and conclude that our method is more accurate in identifying weak spines. Experiments show that our approach can quickly and accurately detect and quantify spines of in vivo neuron images and is able to identify spine elimination and formation. PMID:19434521

  6. Optimum conditions for high-quality 3D reconstruction in confocal scanning microscopy

    NASA Astrophysics Data System (ADS)

    Kim, Taehoon; Kim, Taejoong; Lee, SeungWoo; Gweon, Dae-Gab; Seo, Jungwoo

    2006-02-01

    Confocal Scanning Microscopy (CSM) is very useful to reconstruct 3D image of Bio-cells and the objects that have specification shape in higher axial and lateral resolution and widely used as measurement instrument. A 3D reconstruction is used to visualize confocal images and consists of following processes. The First process is to get 3D data by collecting a series of images at regular focus intervals (Optical Sectioning). The Second process is to fit a curve to a series of 3D data points each pixel. The Third process is to search height information that has maximum value from curve-fitting. However, because of various systematic errors (NOISE) occurred when collecting the information of images through Optical Sectioning and large peak deviation occurred from curve-fitting error, high quality 3D reconstruction is not expected. Also, it takes much time to 3d Reconstruction by using many 3D data in order to acquire high quality and much cost to improve signal-to-noise (SNR) using a higher power laser. So, we are going to define SNR, peak deviation and the order of curve-fitting as important factors and simulate the relation between the factors in order to find a optimum condition for high quality 3D reconstruction in Confoal Scanning Microscopy. If we use optimum condition obtained by this simulation, using a suitable SNR and the suitable number of data and the suitable n-th order curve-fitting, small peak deviation is expected and then, 3D reconstruction of little better quality is expected. Also, it is expected to save.

  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

    2013-05-03

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

  8. 3D geometry-based quantification of colocalizations in multichannel 3D microscopy images of human soft tissue tumors.

    PubMed

    Wörz, Stefan; Sander, Petra; Pfannmöller, Martin; Rieker, Ralf J; Joos, Stefan; Mechtersheimer, Gunhild; Boukamp, Petra; Lichter, Peter; Rohr, Karl

    2010-08-01

    We introduce a new model-based approach for automatic quantification of colocalizations in multichannel 3D microscopy images. The approach uses different 3D parametric intensity models in conjunction with a model fitting scheme to localize and quantify subcellular structures with high accuracy. The central idea is to determine colocalizations between different channels based on the estimated geometry of the subcellular structures as well as to differentiate between different types of colocalizations. A statistical analysis was performed to assess the significance of the determined colocalizations. This approach was used to successfully analyze about 500 three-channel 3D microscopy images of human soft tissue tumors and controls.

  9. Imaging the behavior of molecules in biological systems: breaking the 3D speed barrier with 3D multi-resolution microscopy.

    PubMed

    Welsher, Kevin; Yang, Haw

    2015-01-01

    The overwhelming effort in the development of new microscopy methods has been focused on increasing the spatial and temporal resolution in all three dimensions to enable the measurement of the molecular scale phenomena at the heart of biological processes. However, there exists a significant speed barrier to existing 3D imaging methods, which is associated with the overhead required to image large volumes. This overhead can be overcome to provide nearly unlimited temporal precision by simply focusing on a single molecule or particle via real-time 3D single-particle tracking and the newly developed 3D Multi-resolution Microscopy (3D-MM). Here, we investigate the optical and mechanical limits of real-time 3D single-particle tracking in the context of other methods. In particular, we investigate the use of an optical cantilever for position sensitive detection, finding that this method yields system magnifications of over 3000×. We also investigate the ideal PID control parameters and their effect on the power spectrum of simulated trajectories. Taken together, these data suggest that the speed limit in real-time 3D single particle-tracking is a result of slow piezoelectric stage response as opposed to optical sensitivity or PID control.

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

    PubMed

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

    2013-08-12

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

  11. 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.

  12. High-resolution 3D structural and optical analyses of hybrid or composite materials by means of scanning probe microscopy combined with the ultramicrotome technique: an example of application to engineering of liquid crystals doped with fluorescent quantum dots

    NASA Astrophysics Data System (ADS)

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

    2013-05-01

    Combination of nanometer-scale 3D structural analysis with optical characterization of the same material is a challenging task. Its results may be important for nanophotonics, materials science, and quality control. We have developed a new technique for complementary high-resolution structural and optical characterization followed by optical spectroscopic and microscopic measurements accompanied by reconstruction of the 3D structure in the same area of the sample. The 3D structure is reconstructed by combination of ultramicrotomic and SPM techniques allowing the study of the 3D distribution of implanted nanoparticles and their effect on the matrix structure. The combination of scanning probe nanotomography (SPN) and optical microspectroscopy makes it possible to direct estimate how the 3D structural characteristics of materials affect their macroscopic optical properties. The technique developed has been applied to the engineering of materials made from cholesteric liquid crystals and fluorescent quantum dots (QDs). These materials permit photochemical patterning and image recording through the changes in the dissymmetry factor of circular polarization of QD emission. The differences in the polarisation images and morphological characteristics of the liquid crystal matrix have proved to be correlated with the arrangement of the areas of homogeneous distribution and nonhomogeneous clustering of QDs. The reconstruction of the 3D structure of the liquid crystal matrix in the areas of homogeneous QD distribution has shown that QDs embedded into cholesteric liquid crystal matrices do not perturb their periodic planar texture. The combined optical/SPM/ultramicrotome technique will be indispensable for evaluating the effects of inorganic nanoparticles on the organisation of organic and liquid crystal matrices, biomedical materials, cells, and tissues.

  13. 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.

  14. 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.

  15. Applications of Digital Holography: From Microscopy to 3D-Television

    NASA Astrophysics Data System (ADS)

    Kreis, T.

    2012-03-01

    The paper gives an overview of the applications of digital holography based on the one hand on CCD-recording, computer storage, and numerical reconstruction of the wave fields, and on the other hand on numerical calculation of computer generated holograms (CGH) and the transfer of these CGHs to spatial light modulators (SLM) for optical reconstruction of the wave fields. The first mentioned type of digital holography finds applications in digital holographic microscopy, particle analysis, and interferometric form and deformation measurement, while the second type constitutes the basis for holographic 3D TV. The space-bandwidth-problem occuring in this context is addressed and first partial solutions are presented.

  16. 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.

  17. Investigation of osteoblast cells behavior in polymeric 3D micropatterned scaffolds using digital holographic microscopy.

    PubMed

    Mihailescu, M; Popescu, R C; Matei, A; Acasandrei, A; Paun, I A; Dinescu, M

    2014-08-01

    The effect of micropatterned polymeric scaffolds on the features of the cultured cells at different time intervals after seeding was investigated by digital holographic microscopy. Both parallel and perpendicular walls, with different heights, were fabricated using two-photon lithography on photopolymers. The walls were subsequently coated with polypyrrole-based thin films using the matrix assisted pulsed laser evaporation technique. Osteoblast-like cells, MG-63 line, were cultured on these polymeric 3D micropatterned scaffolds. To analyze these scaffolds with/without cultured cells, an inverted digital holographic microscope, which provides 3D images, was used. Information about the samples' refractive indices and heights was obtained from the phase shift introduced in the optical path. Characteristics of cell adhesion, alignment, orientation, and morphology as a function of the wall heights and time from seeding were highlighted.

  18. Optical 3D imaging and visualization of concealed objects

    NASA Astrophysics Data System (ADS)

    Berginc, G.; Bellet, J.-B.; Berechet, I.; Berechet, S.

    2016-09-01

    This paper gives new insights on optical 3D imagery. In this paper we explore the advantages of laser imagery to form a three-dimensional image of the scene. 3D laser imaging can be used for three-dimensional medical imaging and surveillance because of ability to identify tumors or concealed objects. We consider the problem of 3D reconstruction based upon 2D angle-dependent laser images. The objective of this new 3D laser imaging is to provide users a complete 3D reconstruction of objects from available 2D data limited in number. The 2D laser data used in this paper come from simulations that are based on the calculation of the laser interactions with the different meshed objects of the scene of interest or from experimental 2D laser images. We show that combining the Radom transform on 2D laser images with the Maximum Intensity Projection can generate 3D views of the considered scene from which we can extract the 3D concealed object in real time. With different original numerical or experimental examples, we investigate the effects of the input contrasts. We show the robustness and the stability of the method. We have developed a new patented method of 3D laser imaging based on three-dimensional reflective tomographic reconstruction algorithms and an associated visualization method. In this paper we present the global 3D reconstruction and visualization procedures.

  19. Research of 3D display using anamorphic optics

    NASA Astrophysics Data System (ADS)

    Matsumoto, Kenji; Honda, Toshio

    1997-05-01

    This paper describes the auto-stereoscopic display which can reconstruct more reality and viewer friendly 3-D image by increasing the number of parallaxes and giving motion parallax horizontally. It is difficult to increase number of parallaxes to give motion parallax to the 3-D image without reducing the resolution, because the resolution of display device is insufficient. The magnification and the image formation position can be selected independently in horizontal direction and the vertical direction by projecting between the display device and the 3-D image with the anamorphic optics. The anamorphic optics is an optics system with different magnification in horizontal direction and the vertical direction. It consists of the combination of cylindrical lenses with different focal length. By using this optics, even if we use a dynamic display such as liquid crystal display (LCD), it is possible to display the realistic 3-D image having motion parallax. Motion parallax is obtained by assuming width of the single parallax at the viewing position to be about the same size as the pupil diameter of viewer. In addition, because the focus depth of the 3-D image is deep in this method, conflict of accommodation and convergence is small, and natural 3-D image can be displayed.

  20. 3D reconstruction of cortical microtubules using multi-angle total internal reflection fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Jin, Luhong; Xiu, Peng; Zhou, Xiaoxu; Fan, Jiannan; Kuang, Cuifang; Liu, Xu; Xu, Yingke

    2017-01-01

    Total internal reflection fluorescence microscopy (TIRFM) has been widely used in biomedical research to visualize cellular processes near the cell surface. In this study, a novel multi-angle ring-illuminated TIRFM system, equipped with two galvo mirrors that are on conjugate plan of a 4f optical system was developed. Multi-angle TIRFM generates images with different penetration depths through the controlled variation of the incident angle of illuminating laser. We presented a method to perform three-dimensional (3-D) reconstruction of microtubules from multi-angle TIRFM images. The performance of our method was validated in simulated microtubules with variable signal-to-noise ratios (SNR) and the axial resolution and accuracy of reconstruction were evaluated in selecting different numbers of illumination angles or in different SNR conditions. In U373 cells, we reconstructed the 3-D localization of microtubules near the cell surface with high resolution using over a hundred different illumination angles. Theoretically, the presented TIRFM setup and 3-D reconstruction method can achieve 40 nm axial resolution in experimental conditions where SNR is as low as 2, with 35 different illumination angles. Moreover, our system and reconstruction method have the potential to be used in live cells to track membrane dynamics in 3-D.

  1. 3D simulation for solitons used in optical fibers

    NASA Astrophysics Data System (ADS)

    Vasile, F.; Tebeica, C. M.; Schiopu, P.; Vladescu, M.

    2016-12-01

    In this paper is described 3D simulation for solitions used in optical fibers. In the scientific works is started from nonlinear propagation equation and the solitons represents its solutions. This paper presents the simulation of the fundamental soliton in 3D together with simulation of the second order soliton in 3D. These simulations help in the study of the optical fibers for long distances and in the interactions between the solitons. This study helps the understanding of the nonlinear propagation equation and for nonlinear waves. These 3D simulations are obtained using MATLAB programming language, and we can observe fundamental difference between the soliton and the second order/higher order soliton and in their evolution.

  2. 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.

  3. High-speed optical 3D sensing and its applications

    NASA Astrophysics Data System (ADS)

    Watanabe, Yoshihiro

    2016-12-01

    This paper reviews high-speed optical 3D sensing technologies for obtaining the 3D shape of a target using a camera. The focusing speed is from 100 to 1000 fps, exceeding normal camera frame rates, which are typically 30 fps. In particular, contactless, active, and real-time systems are introduced. Also, three example applications of this type of sensing technology are introduced, including surface reconstruction from time-sequential depth images, high-speed 3D user interaction, and high-speed digital archiving.

  4. 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.

  5. Optical imaging. Expansion microscopy.

    PubMed

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

    2015-01-30

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

  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. Progresses in 3D integral imaging with optical processing

    NASA Astrophysics Data System (ADS)

    Martínez-Corral, Manuel; Martínez-Cuenca, Raúl; Saavedra, Genaro; Navarro, Héctor; Pons, Amparo; Javidi, Bahram

    2008-11-01

    Integral imaging is a promising technique for the acquisition and auto-stereoscopic display of 3D scenes with full parallax and without the need of any additional devices like special glasses. First suggested by Lippmann in the beginning of the 20th century, integral imaging is based in the intersection of ray cones emitted by a collection of 2D elemental images which store the 3D information of the scene. This paper is devoted to the study, from the ray optics point of view, of the optical effects and interaction with the observer of integral imaging systems.

  8. 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.

  9. Optical Approach to Resin Formulation for 3D Printed Microfluidics†

    PubMed Central

    Gong, Hua; Beauchamp, Michael; Perry, Steven; Woolley, Adam T.

    2015-01-01

    Microfluidics imposes different requirements on 3D printing compared to many applications because the critical features for microfluidics consist of internal microvoids. Resins for general 3D printing applications, however, are not necessarily formulated to meet the requirements of microfluidics and minimize the size of fabricated voids. In this paper we use an optical approach to guide custom formulation of resins to minimize the cross sectional size of fabricated flow channels as exemplars of such voids. We focus on stereolithgraphy (SL) 3D printing with Digital Light Processing (DLP) based on a micromirror array and use a commercially available 3D printer. We develop a mathematical model for the optical dose delivered through the thickness of a 3D printed part, including the effect of voids. We find that there is a fundamental trade-off between the homogeneity of the optical dose within individual layers and how far the critical dose penetrates into a flow channel during fabrication. We also experimentally investigate the practical limits of flow channel miniaturization given the optical properties of a resin and find that the minimum flow channel height is ~3.5–5.5ha where ha is the optical penetration depth of the resin, and that the minimum width is 4 pixels in the build plane. We also show that the ratio of the build layer thickness to ha should be in the range 0.3–1.0 to obtain the minimum flow channel height for a given resin. The minimum flow channel size that we demonstrate for a custom resin is 60 μm × 108 μm for a 10 μm build layer thickness. This work lays the foundation for 3D printing of <100 μm microfluidic features. PMID:26744624

  10. 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.

  11. Two-Layer Elastographic 3-D Traction Force Microscopy

    NASA Astrophysics Data System (ADS)

    Álvarez-González, Begoña; Zhang, Shun; Gómez-González, Manuel; Meili, Ruedi; Firtel, Richard A.; Lasheras, Juan C.; Del Álamo, Juan C.

    2017-01-01

    Cellular traction force microscopy (TFM) requires knowledge of the mechanical properties of the substratum where the cells adhere to calculate cell-generated forces from measurements of substratum deformation. Polymer-based hydrogels are broadly used for TFM due to their linearly elastic behavior in the range of measured deformations. However, the calculated stresses, particularly their spatial patterns, can be highly sensitive to the substratum’s Poisson’s ratio. We present two-layer elastographic TFM (2LETFM), a method that allows for simultaneously measuring the Poisson’s ratio of the substratum while also determining the cell-generated forces. The new method exploits the analytical solution of the elastostatic equation and deformation measurements from two layers of the substratum. We perform an in silico analysis of 2LETFM concluding that this technique is robust with respect to TFM experimental parameters, and remains accurate even for noisy measurement data. We also provide experimental proof of principle of 2LETFM by simultaneously measuring the stresses exerted by migrating Physarum amoeboae on the surface of polyacrylamide substrata, and the Poisson’s ratio of the substrata. The 2LETFM method could be generalized to concurrently determine the mechanical properties and cell-generated forces in more physiologically relevant extracellular environments, opening new possibilities to study cell-matrix interactions.

  12. Two-Layer Elastographic 3-D Traction Force Microscopy

    PubMed Central

    Álvarez-González, Begoña; Zhang, Shun; Gómez-González, Manuel; Meili, Ruedi; Firtel, Richard A.; Lasheras, Juan C.; del Álamo, Juan C.

    2017-01-01

    Cellular traction force microscopy (TFM) requires knowledge of the mechanical properties of the substratum where the cells adhere to calculate cell-generated forces from measurements of substratum deformation. Polymer-based hydrogels are broadly used for TFM due to their linearly elastic behavior in the range of measured deformations. However, the calculated stresses, particularly their spatial patterns, can be highly sensitive to the substratum’s Poisson’s ratio. We present two-layer elastographic TFM (2LETFM), a method that allows for simultaneously measuring the Poisson’s ratio of the substratum while also determining the cell-generated forces. The new method exploits the analytical solution of the elastostatic equation and deformation measurements from two layers of the substratum. We perform an in silico analysis of 2LETFM concluding that this technique is robust with respect to TFM experimental parameters, and remains accurate even for noisy measurement data. We also provide experimental proof of principle of 2LETFM by simultaneously measuring the stresses exerted by migrating Physarum amoeboae on the surface of polyacrylamide substrata, and the Poisson’s ratio of the substrata. The 2LETFM method could be generalized to concurrently determine the mechanical properties and cell-generated forces in more physiologically relevant extracellular environments, opening new possibilities to study cell-matrix interactions. PMID:28074837

  13. 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

  14. 3D optical measuring technologies for dimensional inspection

    NASA Astrophysics Data System (ADS)

    Chugui, Yu V.

    2005-01-01

    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, development of hole inspection method on the base of diffractive optical elements. Ensuring the safety of nuclear reactors and running trains as well as their high exploitation reliability takes a noncontact inspection of geometrical parameters of their components. For this tasks we have developed methods and produced the technical vision measuring systems LMM, CONTROL, PROFILE, and technologies for non-contact 3D dimensional inspection of grid spacers and fuel elements for the nuclear reactor VVER-1000 and VVER-440, as well as automatic laser diagnostic system COMPLEX for noncontact inspection of geometrical parameters of running freight car wheel pairs. The performances of these systems and the results of the industrial testing at atomic and railway companies are presented.

  15. 3D optical measuring technologies and systems for industrial applications

    NASA Astrophysics Data System (ADS)

    Chugui, Yu. V.

    2005-06-01

    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, development of hole inspection method on the base of diffractive optical elements. 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, RADAR, 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.

  16. Unity Occupation of Sites in a 3D Optical Lattice

    NASA Astrophysics Data System (ADS)

    Depue, Marshall T.; McCormick, Colin; Winoto, S. Lukman; Oliver, Steven; Weiss, David S.

    1999-03-01

    An average filling factor of one atom per lattice site has been obtained in a submicron scale far-off-resonance optical lattice (FORL). High site occupation is obtained through a compression sequence that includes laser cooling in a 3D FORL and adiabatic toggling between the 3D FORL and a 1D FORL trap. After the highest filling factor is achieved, laser cooling causes collisional loss from lattice sites with more than one atom. Ultimately 44% of the sites have a single atom cooled to near its vibrational ground state. A theoretical model of site occupation based on Poisson statistics agrees well with the experimental results.

  17. Rapid reconstruction of 3D neuronal morphology from light microscopy images with augmented rayburst sampling.

    PubMed

    Ming, Xing; Li, Anan; Wu, Jingpeng; Yan, Cheng; Ding, Wenxiang; Gong, Hui; Zeng, Shaoqun; Liu, Qian

    2013-01-01

    Digital reconstruction of three-dimensional (3D) neuronal morphology from light microscopy images provides a powerful technique for analysis of neural circuits. It is time-consuming to manually perform this process. Thus, efficient computer-assisted approaches are preferable. In this paper, we present an innovative method for the tracing and reconstruction of 3D neuronal morphology from light microscopy images. The method uses a prediction and refinement strategy that is based on exploration of local neuron structural features. We extended the rayburst sampling algorithm to a marching fashion, which starts from a single or a few seed points and marches recursively forward along neurite branches to trace and reconstruct the whole tree-like structure. A local radius-related but size-independent hemispherical sampling was used to predict the neurite centerline and detect branches. Iterative rayburst sampling was performed in the orthogonal plane, to refine the centerline location and to estimate the local radius. We implemented the method in a cooperative 3D interactive visualization-assisted system named flNeuronTool. The source code in C++ and the binaries are freely available at http://sourceforge.net/projects/flneurontool/. We validated and evaluated the proposed method using synthetic data and real datasets from the Digital Reconstruction of Axonal and Dendritic Morphology (DIADEM) challenge. Then, flNeuronTool was applied to mouse brain images acquired with the Micro-Optical Sectioning Tomography (MOST) system, to reconstruct single neurons and local neural circuits. The results showed that the system achieves a reasonable balance between fast speed and acceptable accuracy, which is promising for interactive applications in neuronal image analysis.

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

    PubMed

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

    2016-09-16

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-09-01

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

  20. 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.

  1. Continuous Optical 3D Printing of Green Aliphatic Polyurethanes.

    PubMed

    Pyo, Sang-Hyun; Wang, Pengrui; Hwang, Henry H; Zhu, Wei; Warner, John; Chen, Shaochen

    2017-01-11

    Photosensitive diurethanes were prepared from a green chemistry synthesis pathway based on methacrylate-functionalized six-membered cyclic carbonate and biogenic amines. A continuous optical 3D printing method for the diurethanes was developed to create user-defined gradient stiffness and smooth complex surface microstructures in seconds. The green chemistry-derived polyurethane (gPU) showed high optical transparency, and we demonstrate the ability to tune the material stiffness of the printed structure along a gradient by controlling the exposure time and selecting various amine compounds. High-resolution 3D biomimetic structures with smooth curves and complex contours were printed using our gPU. High cell viability (over 95%) was demonstrated during cytocompatibility testing using C3H 10T1/2 cells seeded directly on the printed structures.

  2. 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

  3. Dual-Color 3D Superresolution Microscopy by Combined Spectral-Demixing and Biplane Imaging

    PubMed Central

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

    2015-01-01

    Multicolor three-dimensional (3D) superresolution techniques allow important insight into the relative organization of cellular structures. While a number of innovative solutions have emerged, multicolor 3D techniques still face significant technical challenges. In this Letter we provide a straightforward approach to single-molecule localization microscopy imaging in three dimensions and two colors. We combine biplane imaging and spectral-demixing, which eliminates a number of problems, including color cross-talk, chromatic aberration effects, and problems with color registration. We present 3D dual-color images of nanoscopic structures in hippocampal neurons with a 3D compound resolution routinely achieved only in a single color. PMID:26153696

  4. 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.

  5. 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

  6. 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.

  7. Total body irradiation with a compensator fabricated using a 3D optical scanner and a 3D printer.

    PubMed

    Park, So-Yeon; Kim, Jung-In; Joo, Yoon Ha; Lee, Jung Chan; Park, Jong Min

    2017-05-07

    We propose bilateral total body irradiation (TBI) utilizing a 3D printer and a 3D optical scanner. We acquired surface information of an anthropomorphic phantom with the 3D scanner and fabricated the 3D compensator with the 3D printer, which could continuously compensate for the lateral missing tissue of an entire body from the beam's eye view. To test the system's performance, we measured doses with optically stimulated luminescent dosimeters (OSLDs) as well as EBT3 films with the anthropomorphic phantom during TBI without a compensator, conventional bilateral TBI, and TBI with the 3D compensator (3D TBI). The 3D TBI showed the most uniform dose delivery to the phantom. From the OSLD measurements of the 3D TBI, the deviations between the measured doses and the prescription dose ranged from  -6.7% to 2.4% inside the phantom and from  -2.3% to 0.6% on the phantom's surface. From the EBT3 film measurements, the prescription dose could be delivered to the entire body of the phantom within  ±10% accuracy, except for the chest region, where tissue heterogeneity is extreme. The 3D TBI doses were much more uniform than those of the other irradiation techniques, especially in the anterior-to-posterior direction. The 3D TBI was advantageous, owing to its uniform dose delivery as well as its efficient treatment procedure.

  8. 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.

  9. 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

  10. 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

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

    PubMed

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

    2013-10-21

    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.

  13. 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

  14. 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.

  15. 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.

  16. Optical characterization of different types of 3D displays

    NASA Astrophysics Data System (ADS)

    Boher, Pierre; Leroux, Thierry; Bignon, Thibault; Collomb-Patton, Véronique

    2012-03-01

    All 3D displays have the same intrinsic method to induce depth perception. They provide different images in the left and right eye of the observer to obtain the stereoscopic effect. The three most common solutions already available on the market are active glass, passive glass and auto-stereoscopic 3D displays. The three types of displays are based on different physical principle (polarization, time selection or spatial emission) and consequently require different measurement instruments and techniques. In the proposed paper, we present some of these solutions and the technical characteristics that can be obtained to compare the displays. We show in particular that local and global measurements can be made in the three cases to access to different characteristics. We also discuss the new technologies currently under development and their needs in terms of optical characterization.

  17. Confocal laser scanning microscopy and 3-D reconstructions of neuronal structures in human brain cortex.

    PubMed

    Belichenko, P V; Dahlström, A

    1995-09-01

    Human brain material was studied with Lucifer yellow (LY) microinjections, indirect Texas red immunofluorescence, and confocal laser scanning microscopy (CLSM). The scanned images were transferred to a Silicon Graphics (SG) IRIS computer equipped with software for reconstructing the 3-D architecture of cells. By employing dual channel CLSM (Bio-Rad MRC 600), LY-injected cells and Texas red immunofluorescence could be studied simultaneously. Autopsy material with 2- to 48-h postmortem delays (6 control and 2 Rett's syndrome cases) as well as biopsy material (14 cases with therapy-resistant partial epilepsy--TRPE--undergoing neurosurgery) were used. In each specimen, 100-200 pyramidal and nonpyramidal neurons were visualized by LY microinjection. Single neurons were imaged and 2-D reconstructions of each neuron were made using z-projections of serial optical images; 3-D reconstructions and rotations were computed using the SG workstation, with VoxelView software from Vital Images (UK), and stored in a "neuronal library" on laser or magnetic optical disks. In Ret's syndrome cases and in patients with TRPE various abnormalities in the dendritic geometry of pyramidal and nonpyramidal cells have been found. The combination of LY injections with immunofluorescence allows the investigation of transmitter-related substances around the LY-injected cells. Using antibodies to synaptic vesicle proteins, presynaptic elements docking onto individual spines have been demonstrated. This approach may contribute to the understanding of different neurological and psychiatric disorders and may be useful in the Mapping of the Human Brain project. It may also be integrated with functional imaging by PET scan and with the human genome project.

  18. 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.

  19. Extraterrestrial optical microscopy.

    PubMed

    Soffen, G A

    1969-07-01

    An examination of the literature concerned with the use of microscopy for planetary investigation reveals a serious deficiency of current efforts. Many scientists have recommended the use of a microscope for planetary investigation [Biology and the Exploration of Mars, C. S. Pittendrigh, W. Vishniac, and J. P. T. Pearman, Eds. (National Academy of Science-National Research Council, Washington, D. C., 1966), (a) D. Mazia, p. 31; (b) J. Lederberg, p. 137; (c) S. Fox, pp. 219, 226; (d) D. Glaser, p. 326; (e) D. Glaser, J. McCarthy, and M. Minsky, pp. 333, 341; (f) D. G. Rea, pp. 347-426; (g) P. G. Conger, pp. 409-414; (h) M. H. Fernandez, pp. 414-425; (i) D. Schwartz, pp.425-426 . H. P. Klein, Some Biological Problems in the Search for Extraterrestrial Life (American Astronautical Society, Washington, D. C., 1968).] but few are involved in developing the experiment. Since this is a particularly timely period for the preparation of planetary lander experiments, the reasons for this lack of effort would appear to be limited resources or an unclear course of action, rather than lack of interest. Microscopy used for planetary investigation is chiefly the interest of the biologist and the mineralogist. In both cases the desire to use magnifying optics in order to observe objects of submillimeter size is based upon the rich body of knowledge we have acquired from observing the terrestrial microcosm. In addition to purely imaging, certain special optical techniques, e.g., polarimetry, colorimetry, phase contrast, etc., can be used to enhance the interpretation of microscopic imaging data. This interaction of the optical with the chemical or structural aspects of nature can be used to great advantage in the exploration of extraterrestrial biology and mineralogy.

  20. Structural and functional imaging of 3D microfluidic mixers using optical coherence tomography.

    PubMed

    Xi, Chuanwu; Marks, Daniel L; Parikh, Devang S; Raskin, Lutgarde; Boppart, Stephen A

    2004-05-18

    To achieve high mixing efficiency in microfluidic devices, complex designs are often required. Microfluidic devices have been evaluated with light and confocal microscopy, but fluid-flow characteristics at different depths are difficult to separate from the en face images produced. By using optical coherence tomography (OCT), an imaging modality capable of imaging 3D microstructures at micrometer-scale resolutions over millimeter-size scales, we obtained 3D dynamic functional and structural data for three representative microfluidic mixers: a Y channel mixer, a 3D serpentine mixer, and a vortex mixer. In the serpentine mixer, OCT image analysis revealed that the mixing efficiency was linearly dependent on the Reynolds number, whereas it appeared to have exponential dependence when imaged with light microscopy. The visual overlap of fluid flows in light-microscopy images leads to an overestimation of the mixing efficiency, an effect that was eliminated with OCT imaging. Doppler OCT measurements determined velocity profiles at various points in the serpentine mixer. Mixing patterns in the vortex mixer were compared with light-microscopy and OCT image analysis. These results demonstrate that OCT can significantly improve the characterization of 3D microfluidic device structure and function.

  1. Ultracold Heteronuclear Molecules in a 3D Optical Lattice

    SciTech Connect

    Ospelkaus, C.; Ospelkaus, S.; Humbert, L.; Ernst, P.; Sengstock, K.; Bongs, K.

    2006-09-22

    We report on the creation of ultracold heteronuclear molecules assembled from fermionic {sup 40}K and bosonic {sup 87}Rb atoms in a 3D optical lattice. Molecules are produced at a heteronuclear Feshbach resonance on both the attractive and the repulsive sides of the resonance. We precisely determine the binding energy of the heteronuclear molecules from rf spectroscopy across the Feshbach resonance. We characterize the lifetime of the molecular sample as a function of magnetic field and measure lifetimes between 20 and 120 ms. The efficiency of molecule creation via rf association is measured and is found to decrease as expected for more deeply bound molecules.

  2. 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.

  3. 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.

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

    PubMed

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

    2013-10-22

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

  5. Quantitative IR microscopy and spectromics open the way to 3D digital pathology.

    PubMed

    Bobroff, Vladimir; Chen, Hsiang-Hsin; Delugin, Maylis; Javerzat, Sophie; Petibois, Cyril

    2016-06-01

    Currently, only mass-spectrometry (MS) microscopy brings a quantitative analysis of chemical contents of tissue samples in 3D. Here, the reconstruction of a 3D quantitative chemical images of a biological tissue by FTIR spectro-microscopy is reported. An automated curve-fitting method is developed to extract all intense absorption bands constituting IR spectra. This innovation benefits from three critical features: (1) the correction of raw IR spectra to make them quantitatively comparable; (2) the automated and iterative data treatment allowing to transfer the IR-absorption spectrum into a IR-band spectrum; (3) the reconstruction of an 3D IR-band matrix (x, y, z for voxel position and a 4(th) dimension with all IR-band parameters). Spectromics, which is a new method for exploiting spectral data for tissue metadata reconstruction, is proposed to further translate the related chemical information in 3D, as biochemical and anatomical tissue parameters. An example is given with oxidative stress distribution and the reconstruction of blood vessels in tissues. The requirements of IR microscopy instrumentation to propose 3D digital histology as a clinical routine technology is briefly discussed.

  6. Novel scanning electron microscopy methods for analyzing the 3D structure of the Golgi apparatus.

    PubMed

    Koga, Daisuke; Ushiki, Tatsuo; Watanabe, Tsuyoshi

    2017-01-01

    The structure of the Golgi apparatus has been extensively examined by light and electron microscopy, but details of its three-dimensional (3D) structure have remained unclear because of the technical limitations of conventional microscopy techniques. To overcome this problem, we have developed several novel scanning electron microscopy (SEM) methods for observing the 3D structure of subcellular organelles including the Golgi apparatus: (1) an osmium maceration method that facilitates SEM observation of membranous organelles, including the Golgi apparatus, by selectively removing soluble cytoplasmic proteins, (2) an osmium impregnation/maceration method that combines an osmium impregnation method with the osmium maceration method to determine the polarity of the Golgi apparatus by SEM, (3) a correlative light and SEM method that combines a cryosectioning technique with the osmium maceration method to enable correlation of the immunocytochemical distribution of molecules with the 3D ultrastructure of the Golgi apparatus, and (4) array tomography based on the systematic collection and integration of SEM images of serial ultrathin sections on glass slides for revealing the 3D ultrastructure of the entire Golgi apparatus. Together, the novel SEM techniques listed above can reveal the complete 3D structure of the Golgi apparatus in different cell types.

  7. 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

  8. 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.

  9. Ultracold polar molecules in a 3D optical lattice

    NASA Astrophysics Data System (ADS)

    Yan, Bo

    2015-05-01

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

  10. 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.

  11. 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

  12. 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

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

    PubMed

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

    2015-07-01

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

  14. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

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

  16. Staining and embedding of human chromosomes for 3-d serial block-face scanning electron microscopy.

    PubMed

    Yusuf, Mohammed; Chen, Bo; Hashimoto, Teruo; Estandarte, Ana Katrina; Thompson, George; Robinson, Ian

    2014-12-01

    The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial block-face scanning electron microscopy (SBFSEM). Polyamine chromosomes are first separated using a simple filtration method and then stained with heavy metal. We show that the DNA-specific platinum blue provides higher contrast than osmium tetroxide. A two-step procedure for embedding chromosomes in resin is then used to concentrate the chromosome samples. After stacking the SBFSEM images, a familiar X-shaped chromosome was observed in 3-D.

  17. New data-driven method from 3D confocal microscopy for calculating phytoplankton cell biovolume.

    PubMed

    Roselli, L; Paparella, F; Stanca, E; Basset, A

    2015-06-01

    Confocal laser scanner microscopy coupled with an image analysis system was used to directly determine the shape and calculate the biovolume of phytoplankton organisms by constructing 3D models of cells. The study was performed on Biceratium furca (Ehrenberg) Vanhoeffen, which is one of the most complex-shaped phytoplankton. Traditionally, biovolume is obtained from a standardized set of geometric models based on linear dimensions measured by light microscopy. However, especially in the case of complex-shaped cells, biovolume is affected by very large errors associated with the numerous manual measurements that this entails. We evaluate the accuracy of these traditional methods by comparing the results obtained using geometric models with direct biovolume measurement by image analysis. Our results show cell biovolume measurement based on decomposition into simple geometrical shapes can be highly inaccurate. Although we assume that the most accurate cell shape is obtained by 3D direct biovolume measurement, which is based on voxel counting, the intrinsic uncertainty of this method is explored and assessed. Finally, we implement a data-driven formula-based approach to the calculation of biovolume of this complex-shaped organism. On one hand, the model is obtained from 3D direct calculation. On the other hand, it is based on just two linear dimensions which can easily be measured by hand. This approach has already been used for investigating the complexities of morphology and for determining the 3D structure of cells. It could also represent a novel way to generalize scaling laws for biovolume calculation.

  18. Photometry unlocks 3D information from 2D localization microscopy data.

    PubMed

    Franke, Christian; Sauer, Markus; van de Linde, Sebastian

    2017-01-01

    We developed a straightforward photometric method, temporal, radial-aperture-based intensity estimation (TRABI), that allows users to extract 3D information from existing 2D localization microscopy data. TRABI uses the accurate determination of photon numbers in different regions of the emission pattern of single emitters to generate a z-dependent photometric parameter. This method can determine fluorophore positions up to 600 nm from the focal plane and can be combined with biplane detection to further improve axial localization.

  19. Using 3D Super-Resolution Microscopy to Probe Breast Cancer Stem Cells and Their Microenvironment

    DTIC Science & Technology

    2014-05-01

    microenvironments on breast cancer by creating arrays of polydimethlysiloxane (PDMS) microposts of different stiffness and sizes and seeded them with MCF-7 cells...of MCF-7s. Finally, with QPI, we investigated the real-time response of breast- cancer cells to different microenvironmental cues . We thus have...controls this cellular phenotype. To realize this goal, we had proposed to use 3D super-resolution microscopy to visualize how individual breast CaSCs

  20. Quantitative analyses of the 3D nuclear landscape recorded with super-resolved fluorescence microscopy.

    PubMed

    Schmid, Volker J; Cremer, Marion; Cremer, Thomas

    2017-03-18

    Recent advancements of super-resolved fluorescence microscopy have revolutionized microscopic studies of cells, including the exceedingly complex structural organization of cell nuclei in space and time. In this paper we describe and discuss tools for (semi-) automated, quantitative 3D analyses of the spatial nuclear organization. These tools allow the quantitative assessment of highly resolved different chromatin compaction levels in individual cell nuclei, which reflect functionally different regions or sub-compartments of the 3D nuclear landscape, and measurements of absolute distances between sites of different chromatin compaction. In addition, these tools allow 3D mapping of specific DNA/RNA sequences and nuclear proteins relative to the 3D chromatin compaction maps and comparisons of multiple cell nuclei. The tools are available in the free and open source R packages nucim and bioimagetools. We discuss the use of masks for the segmentation of nuclei and the use of DNA stains, such as DAPI, as a proxy for local differences in chromatin compaction. We further discuss the limitations of 3D maps of the nuclear landscape as well as problems of the biological interpretation of such data.

  1. All-in-one 3D printed microscopy chamber for multidimensional imaging, the UniverSlide.

    PubMed

    Alessandri, Kevin; Andrique, Laetitia; Feyeux, Maxime; Bikfalvi, Andreas; Nassoy, Pierre; Recher, Gaëlle

    2017-02-10

    While live 3D high resolution microscopy techniques are developing rapidly, their use for biological applications is partially hampered by practical difficulties such as the lack of a versatile sample chamber. Here, we propose the design of a multi-usage observation chamber adapted for live 3D bio-imaging. We show the usefulness and practicality of this chamber, which we named the UniverSlide, for live imaging of two case examples, namely multicellular systems encapsulated in sub-millimeter hydrogel shells and zebrafish larvae. We also demonstrate its versatility and compatibility with all microscopy devices by using upright or inverted microscope configurations after loading the UniverSlide with fixed or living samples. Further, the device is applicable for medium/high throughput screening and automatized multi-position image acquisition, providing a constraint-free but stable and parallelized immobilization of the samples. The frame of the UniverSlide is fabricated using a stereolithography 3D printer, has the size of a microscopy slide, is autoclavable and sealed with a removable lid, which makes it suitable for use in a controlled culture environment. We describe in details how to build this chamber and we provide all the files necessary to print the different pieces in the lab.

  2. All-in-one 3D printed microscopy chamber for multidimensional imaging, the UniverSlide

    PubMed Central

    Alessandri, Kevin; Andrique, Laetitia; Feyeux, Maxime; Bikfalvi, Andreas; Nassoy, Pierre; Recher, Gaëlle

    2017-01-01

    While live 3D high resolution microscopy techniques are developing rapidly, their use for biological applications is partially hampered by practical difficulties such as the lack of a versatile sample chamber. Here, we propose the design of a multi-usage observation chamber adapted for live 3D bio-imaging. We show the usefulness and practicality of this chamber, which we named the UniverSlide, for live imaging of two case examples, namely multicellular systems encapsulated in sub-millimeter hydrogel shells and zebrafish larvae. We also demonstrate its versatility and compatibility with all microscopy devices by using upright or inverted microscope configurations after loading the UniverSlide with fixed or living samples. Further, the device is applicable for medium/high throughput screening and automatized multi-position image acquisition, providing a constraint-free but stable and parallelized immobilization of the samples. The frame of the UniverSlide is fabricated using a stereolithography 3D printer, has the size of a microscopy slide, is autoclavable and sealed with a removable lid, which makes it suitable for use in a controlled culture environment. We describe in details how to build this chamber and we provide all the files necessary to print the different pieces in the lab. PMID:28186188

  3. All-in-one 3D printed microscopy chamber for multidimensional imaging, the UniverSlide

    NASA Astrophysics Data System (ADS)

    Alessandri, Kevin; Andrique, Laetitia; Feyeux, Maxime; Bikfalvi, Andreas; Nassoy, Pierre; Recher, Gaëlle

    2017-02-01

    While live 3D high resolution microscopy techniques are developing rapidly, their use for biological applications is partially hampered by practical difficulties such as the lack of a versatile sample chamber. Here, we propose the design of a multi-usage observation chamber adapted for live 3D bio-imaging. We show the usefulness and practicality of this chamber, which we named the UniverSlide, for live imaging of two case examples, namely multicellular systems encapsulated in sub-millimeter hydrogel shells and zebrafish larvae. We also demonstrate its versatility and compatibility with all microscopy devices by using upright or inverted microscope configurations after loading the UniverSlide with fixed or living samples. Further, the device is applicable for medium/high throughput screening and automatized multi-position image acquisition, providing a constraint-free but stable and parallelized immobilization of the samples. The frame of the UniverSlide is fabricated using a stereolithography 3D printer, has the size of a microscopy slide, is autoclavable and sealed with a removable lid, which makes it suitable for use in a controlled culture environment. We describe in details how to build this chamber and we provide all the files necessary to print the different pieces in the lab.

  4. Optical 3D sensor for large objects in industrial application

    NASA Astrophysics Data System (ADS)

    Kuhmstedt, Peter; Heinze, Matthias; Himmelreich, Michael; Brauer-Burchardt, Christian; Brakhage, Peter; Notni, Gunther

    2005-06-01

    A new self calibrating optical 3D measurement system using fringe projection technique named "kolibri 1500" is presented. It can be utilised to acquire the all around shape of large objects. The basic measuring principle is the phasogrammetric approach introduced by the authors /1, 2/. The "kolibri 1500" consists of a stationary system with a translation unit for handling of objects. Automatic whole body measurement is achieved by using sensor head rotation and changeable object position, which can be done completely computer controlled. Multi-view measurement is realised by using the concept of virtual reference points. In this way no matching procedures or markers are necessary for the registration of the different images. This makes the system very flexible to realise different measurement tasks. Furthermore, due to self calibrating principle mechanical alterations are compensated. Typical parameters of the system are: the measurement volume extends from 400 mm up to 1500 mm max. length, the measurement time is between 2 min for 12 images up to 20 min for 36 images and the measurement accuracy is below 50μm.The flexibility makes the measurement system useful for a wide range of applications such as quality control, rapid prototyping, design and CAD/CAM which will be shown in the paper.

  5. 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.

  6. Optical tweezers for confocal microscopy

    NASA Astrophysics Data System (ADS)

    Hoffmann, A.; Meyer zu Hörste, G.; Pilarczyk, G.; Monajembashi, S.; Uhl, V.; Greulich, K. O.

    2000-11-01

    In confocal laser scanning microscopes (CLSMs), lasers can be used for image formation as well as tools for the manipulation of microscopic objects. In the latter case, in addition to the imaging lasers, the light of an extra laser has to be focused into the object plane of the CLSM, for example as optical tweezers. Imaging as well as trapping by optical tweezers can be done using the same objective lens. In this case, z-sectioning for 3D imaging shifts the optical tweezers with the focal plane of the objective along the optical axis, so that a trapped object remains positioned in the focal plane. Consequently, 3D imaging of trapped objects is impossible without further measures. We present an experimental set-up keeping the axial trapping position of the optical tweezers at its intended position whilst the focal plane can be axially shifted over a distance of about 15 μm. It is based on fast-moving correctional optics synchronized with the objective movement. First examples of application are the 3D imaging of chloroplasts of Elodea densa (Canadian waterweed) in a vigorous cytoplasmic streaming and the displacement of zymogen granules in pancreatic cancer cells (AR42 J).

  7. Two-Photon Microscopy Analysis of Gold Nanoparticle Uptake in 3D Cell Spheroids

    PubMed Central

    Rane, Tushar D.; Armani, Andrea M.

    2016-01-01

    Nanomaterials can be synthesized from a wide range of material systems in numerous morphologies, creating an extremely diverse portfolio. As result of this tunability, these materials are emerging as a new class of nanotherapeutics and imaging agents. One particularly interesting nanomaterial is the gold nanoparticle. Due to its inherent biocompatibility and tunable photothermal behavior, it has made a rapid transition from the lab setting to in vivo testing. In most nanotherapeutic applications, the efficacy of the agent is directly related to the target of interest. However, the optimization of the AuNP size and shape for efficacy in vitro, prior to testing in in vivo models of a disease, has been largely limited to two dimensional monolayers of cells. Two dimensional cell cultures are unable to reproduce conditions experienced by AuNP in the body. In this article, we systematically investigate the effect of different properties of AuNP on the penetration depth into 3D cell spheroids using two-photon microscopy. The 3D spheroids are formed from the HCT116 cell line, a colorectal carcinoma cell line. In addition to studying different sizes and shapes of AuNPs, we also study the effect of an oligo surface chemistry. There is a significant difference between AuNP uptake profiles in the 2D monolayers of cells as compared to the 3D cell spheroids. Additionally, the range of sizes and shapes studied here also exhibit marked differences in uptake penetration depth and efficacy. Finally, our results demonstrate that two-photon microscopy enables quantitative AuNP localization and concentration data to be obtained at the single spheroid level without fluorescent labeling of the AuNP, thus, providing a viable technique for large scale screening of AuNP properties in 3D cell spheroids as compared to tedious and time consuming techniques like electron microscopy. PMID:27936027

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

    NASA Astrophysics Data System (ADS)

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

    2012-03-01

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

  9. 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

  10. Point scanning confocal microscopy facilitates 3D human hair follicle imaging in tissue sections.

    PubMed

    Kloepper, Jennifer E; Bíró, Tamás; Paus, Ralf; Cseresnyés, Zoltán

    2010-07-01

    Efficiency is a key factor in determining whether a scientific method becomes widely accepted in practical applications. In dermatology, morphological characterisation of intact hair follicles by traditional methods can be rather inefficient. Samples are embedded, sliced, imaged and digitally reconstructed, which can be time-consuming. Confocal microscopy, on the other hand, is more efficient and readily applicable to study intact hair follicles. Modern confocal microscopes deliver and collect light very efficiently and thus allow high spatial resolution imaging of relatively thick samples. In this letter, we report that we successfully imaged entire intact human hair follicles using point scanning confocal microscopy. Light delivery and light-collection were further improved by preparing the samples in 2,2'-Thiodiethanol (TDE), thus reducing refractive index gradients. The relatively short total scan times and the high quality of the acquired 3D images make confocal microscopy a desirable method for studying intact hair follicles under normal and pathological conditions.

  11. Optical microscopy aims deep

    NASA Astrophysics Data System (ADS)

    Gigan, Sylvain

    2017-01-01

    A new set of imaging techniques that take advantage of scattered light may soon lead to key advances in biomedical optics, providing access to depths well beyond what is currently possible with ballistic light.

  12. Hydrothermal synthesis, characterization and optical properties of 3D flower like indium sulfide nanostructures

    NASA Astrophysics Data System (ADS)

    Ghaderi Sheikhi abadi, Parvaneh; Salavati-Niasari, Masoud; Davar, Fatemeh

    2013-01-01

    High-quality and high-yield 3D flower like indium sulfide (In2S3) nanostructures with cubic structure were synthesized by a wet chemical route, without using any surfactant and organic solvents at 160 °C for 12 h, by using InCl3 and 2-aminothiophenol (2-ATP) as starting reagents. The obtained In2S3 with different morphologies and size was characterized by X-ray diffraction pattern (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), and ultraviolet-visible (UV-vis) spectroscopy. The effects of reaction parameters, such as temperature, precursor concentration and reaction time on the morphology, and particle size of products were investigated. Our experimental results showed that temperature and time reaction played key roles in the final morphology of In2S3. The morphology of In2S3 structures could be changed from one-dimensional (1D) structures to three-dimensional (3D) structures by increasing reaction time to 24 h. In the present study the optical properties 3D In2S3 structures were investigated.

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

    PubMed

    Esquivel-Rodríguez, Juan; Kihara, Daisuke

    2013-10-01

    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 3Å. 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.

  14. 3D structure tensor analysis of light microscopy data for validating diffusion MRI

    PubMed Central

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

    2015-01-01

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

  15. Utilization of 3D printing for an intravital microscopy platform to study the intestinal microcirculation.

    PubMed

    Burkovskiy, I; Lehmann, C; Jiang, C; Zhou, J

    2016-11-01

    Intravital microscopy of the intestine is a sophisticated technique that allows qualitative and quantitative in vivo observation of dynamic cellular interactions and blood flow at a high resolution. Physiological conditions of the animal and in particular of the observed organ, such as temperature and moisture are crucial for intravital imaging. Often, the microscopy stage with the animal or the organ of interest imposes limitations on how well the animal can be maintained. In addition, the access for additional oxygen supply or drug administration during the procedure is rather restricted. To address these limitations, we developed a novel intravital microscopy platform, allowing us to have improved access to the animal during the intravital microscopy procedure, as well as improved microenvironmental maintenance. The production process of this prototype platform is based on 3D printing of device parts in a single-step process. The simplicity of production and the advantages of this versatile and customizable design are shown and discussed in this paper. Our design potentially represents a major step forward in facilitating intestinal intravital imaging using fluorescent microscopy.

  16. 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

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

    PubMed

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

    2015-09-01

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

  18. X-ray Laue Diffraction Microscopy in 3D at the Advanced Photon Source

    SciTech Connect

    Liu, W.; Zschack, P.; Tischler, Jonathan Zachary; Ice, Gene E; Larson, Ben C

    2011-01-01

    Studies of materials on mesoscopic length-scales require a penetrating structural probe with submicron point-to-point spatial resolution. The principle research activities at beamline 34-ID-E of the Advanced Photon Source (APS) involve development of exciting new micro-/nano-diffraction techniques for characterization and microscopy in support of both applied engineering and fundamental materials research. Taking advantage of the high brightness of the source, advanced focusing mirrors, a novel depth profiling technique, and high-speed area detectors, three-dimensional scanning Laue diffraction microscopy provides detailed local structural information of crystalline materials, such as crystallographic orientation, orientation gradients, and strain tensors. It is general and applicable to single-crystal, polycrystalline, composite, deformed, and functionally graded materials. Applications include 3D diffraction investigations for a diverse and growing user community with interests in materials deformation, electro-migration, recrystallization, fatigue, solid-solution precipitation, high-pressure environments, and condensed matter physics.

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

    PubMed

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

    2015-05-20

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

  20. 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.

  1. 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.

  2. 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.

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

    PubMed Central

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

    2009-01-01

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

  4. 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.

  5. 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.

  6. Comparison of 3D Orientation Distribution Functions Measured with Confocal Microscopy and Diffusion MRI

    PubMed Central

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

    2016-01-01

    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 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

  7. 3D-confocal microscopy for surface analysis of microstructured materials

    NASA Astrophysics Data System (ADS)

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

    2002-06-01

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

  8. Fast 3-D temporal focusing microscopy using an electrically tunable lens.

    PubMed

    Jiang, Jun; Zhang, Dapeng; Walker, Steven; Gu, Chenglin; Ke, Ya; Yung, Wing Ho; Chen, Shih-chi

    2015-09-21

    In this paper, we present a 3-D temporal focusing microscope based on an electrically tunable lens (ETL) and a femtosecond regenerative laser amplifier. The focus-tunable lens provides a fast and compact way to perform non-mechanical z-scanning and resolves the blurry image issue compared with GVD-based z-scanning methods. The optical performance of the temporal focusing system, including z-scanning characteristics, the associated the magnification variation, and the lateral and axial resolution, has been studied and characterized using calibrated Rhodamine-6G thin film sample, fluorescent beads, and pollen samples. Lastly, we demonstrate the optical cross-sectioning and z-scanning capability with an in vivo experiment, where Ca(2+) imaging of neurons in GaCamp6 labeled zebrafish was performed.

  9. 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.

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

    PubMed

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

    2010-01-01

    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.

  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. 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

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

    PubMed

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

    2015-01-01

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

  14. Automatic segmentation and analysis of fibrin networks in 3D confocal microscopy images

    NASA Astrophysics Data System (ADS)

    Liu, Xiaomin; Mu, Jian; Machlus, Kellie R.; Wolberg, Alisa S.; Rosen, Elliot D.; Xu, Zhiliang; Alber, Mark S.; Chen, Danny Z.

    2012-02-01

    Fibrin networks are a major component of blood clots that provides structural support to the formation of growing clots. Abnormal fibrin networks that are too rigid or too unstable can promote cardiovascular problems and/or bleeding. However, current biological studies of fibrin networks rarely perform quantitative analysis of their structural properties (e.g., the density of branch points) due to the massive branching structures of the networks. In this paper, we present a new approach for segmenting and analyzing fibrin networks in 3D confocal microscopy images. We first identify the target fibrin network by applying the 3D region growing method with global thresholding. We then produce a one-voxel wide centerline for each fiber segment along which the branch points and other structural information of the network can be obtained. Branch points are identified by a novel approach based on the outer medial axis. Cells within the fibrin network are segmented by a new algorithm that combines cluster detection and surface reconstruction based on the α-shape approach. Our algorithm has been evaluated on computer phantom images of fibrin networks for identifying branch points. Experiments on z-stack images of different types of fibrin networks yielded results that are consistent with biological observations.

  15. 3D tracking the Brownian motion of colloidal particles using digital holographic microscopy and joint reconstruction.

    PubMed

    Verrier, Nicolas; Fournier, Corinne; Fournel, Thierry

    2015-06-01

    In-line digital holography is a valuable tool for sizing, locating, and tracking micro- or nano-objects in a volume. When a parametric imaging model is available, inverse problem approaches provide a straightforward estimate of the object parameters by fitting data with the model, thereby allowing accurate reconstruction. As recently proposed and demonstrated, combining pixel super-resolution techniques with inverse problem approaches improves the estimation of particle size and 3D position. Here, we demonstrate the accurate tracking of colloidal particles in Brownian motion. Particle size and 3D position are jointly optimized from video holograms acquired with a digital holographic microscopy setup based on a low-end microscope objective (×20, NA 0.5). Exploiting information redundancy makes it possible to characterize particles with a standard deviation of 15 nm in size and a theoretical resolution of 2×2×5  nm3 for position under additive white Gaussian noise assumption.

  16. 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.

  17. 3D Analysis of Porosity in a Ceramic Coating Using X-ray Microscopy

    NASA Astrophysics Data System (ADS)

    Klement, Uta; Ekberg, Johanna; Kelly, Stephen T.

    2017-02-01

    Suspension plasma spraying (SPS) is a new, innovative plasma spray technique using a feedstock consisting of fine powder particles suspended in a liquid. Using SPS, ceramic coatings with columnar microstructures have been produced which are used as topcoats in thermal barrier coatings. The microstructure contains a wide pore size range consisting of inter-columnar spacings, micro-pores and nano-pores. Hence, determination of total porosity and pore size distribution is a challenge. Here, x-ray microscopy (XRM) has been applied for describing the complex pore space of the coatings because of its capability to image the (local) porosity within the coating in 3D at a resolution down to 50 nm. The possibility to quantitatively segment the analyzed volume allows analysis of both open and closed porosity. For an yttria-stabilized zirconia coating with feathery microstructure, both open and closed porosity were determined and it could be revealed that 11% of the pore volumes (1.4% of the total volume) are closed pores. The analyzed volume was reconstructed to illustrate the distribution of open and closed pores in 3D. Moreover, pore widths and pore volumes were determined. The results on the complex pore space obtained by XRM are discussed in connection with other porosimetry techniques.

  18. Fluorescence fluctuation microscopy to reveal 3D architecture and function in the cell nucleus.

    PubMed

    Lenser, Thorsten; Weisshart, Klaus; Ulbricht, Tobias; Klement, Karolin; Hemmerich, Peter

    2010-01-01

    The three-dimensional (3D) architecture of the cell nucleus is determined not only by the presence of subnuclear domains, such as the nuclear envelope, chromosome territories, and nuclear bodies, but also by smaller domains which form in response to specific functions, such as RNA transcription, DNA replication, and DNA repair. Since both stable and dynamic structures contribute to nuclear morphology, it is important to study the biophysical principles of the formation of macromolecular assemblies within the nucleus. For this purpose, a variety of fluorescence fluctuation microscopy techniques can be applied. Here, we summarize our current knowledge on the 3D architecture of the mammalian cell nucleus and describe in detail how the assembly of functional nuclear protein complexes can be analyzed in living cells using fluorescence bleaching techniques, fluorescence correlation spectroscopy, raster image correlation spectroscopy, and mathematical modeling. In conclusion, the application of all these techniques in combination is a powerful tool to assess the full spectrum of nuclear protein dynamics and to understand the biophysical principles underlying nuclear structure and function.

  19. Automated Atom-By-Atom Three-Dimensional (3D) Reconstruction of Field Ion Microscopy Data.

    PubMed

    Dagan, Michal; Gault, Baptiste; Smith, George D W; Bagot, Paul A J; Moody, Michael P

    2017-03-20

    An automated procedure has been developed for the reconstruction of field ion microscopy (FIM) data that maintains its atomistic nature. FIM characterizes individual atoms on the specimen's surface, evolving subject to field evaporation, in a series of two-dimensional (2D) images. Its unique spatial resolution enables direct imaging of crystal defects as small as single vacancies. To fully exploit FIM's potential, automated analysis tools are required. The reconstruction algorithm developed here relies on minimal assumptions and is sensitive to atomic coordinates of all imaged atoms. It tracks the atoms across a sequence of images, allocating each to its respective crystallographic plane. The result is a highly accurate 3D lattice-resolved reconstruction. The procedure is applied to over 2000 tungsten atoms, including ion-implanted planes. The approach is further adapted to analyze carbides in a steel matrix, demonstrating its applicability to a range of materials. A vast amount of information is collected during the experiment that can underpin advanced analyses such as automated detection of "out of sequence" events, subangstrom surface displacements and defects effects on neighboring atoms. These analyses have the potential to reveal new insights into the field evaporation process and contribute to improving accuracy and scope of 3D FIM and atom probe characterization.

  20. A toolbox for ab initio 3-D reconstructions in single-particle electron microscopy.

    PubMed

    Voss, Neil R; Lyumkis, Dmitry; Cheng, Anchi; Lau, Pick-Wei; Mulder, Anke; Lander, Gabriel C; Brignole, Edward J; Fellmann, Denis; Irving, Christopher; Jacovetty, Erica L; Leung, Albert; Pulokas, James; Quispe, Joel D; Winkler, Hanspeter; Yoshioka, Craig; Carragher, Bridget; Potter, Clinton S

    2010-03-01

    Structure determination of a novel macromolecular complex via single-particle electron microscopy depends upon overcoming the challenge of establishing a reliable 3-D reconstruction using only 2-D images. There are a variety of strategies that deal with this issue, but not all of them are readily accessible and straightforward to use. We have developed a "toolbox" of ab initio reconstruction techniques that provide several options for calculating 3-D volumes in an easily managed and tightly controlled work-flow that adheres to standard conventions and formats. This toolbox is designed to streamline the reconstruction process by removing the necessity for bookkeeping, while facilitating transparent data transfer between different software packages. It currently includes procedures for calculating ab initio reconstructions via random or orthogonal tilt geometry, tomograms, and common lines, all of which have been tested using the 50S ribosomal subunit. Our goal is that the accessibility of multiple independent reconstruction algorithms via this toolbox will improve the ease with which models can be generated, and provide a means of evaluating the confidence and reliability of the final reconstructed map.

  1. Analysis of incomplete excisions of basal-cell carcinomas after breadloaf microscopy compared with 3D-microscopy: a prospective randomized and blinded study.

    PubMed

    Boehringer, Alexandra; Adam, Patrick; Schnabl, Saskia; Häfner, Hans-Martin; Breuninger, Helmut

    2015-08-01

    Basal-cell carcinomas may show irregular, asymmetric subclinical growth. This study analyzed the efficacy of 'breadloaf' microscopy (serial sectioning) and three-dimensional (3D) microscopy in detecting positive tumor margins. Two hundred eighty-three (283) tumors (51.2%) were put into the breadloaf microscopy group; 270 tumors (48.8%) into the 3D microscopy group. The position of any detected tumor outgrowths was identified in clock face fashion. The time required for cutting and embedding the specimens and the examination of the microscopic slides was measured. Patient/tumor characteristics and surgical margins did not differ significantly. Tumor outgrowths at the excision margin were found in 62 of 283 cases (21.9%) in the breadloaf microscopy group and in 115 of 270 cases (42.6%) in the 3D microscopy group, constituting a highly significant difference (p < 0.001). This difference held true with incomplete excision of fibrosing (infiltrative/sclerosing/morpheaform) tumors [32.9% in the breadloaf microscopy group and 57.5% in the 3D microscopy group (p = 0.003)] and also with solid (nodular) tumors [16.1 and 34.2%, respectively (p < 0.001)]. The mean overall examination time required showed no important difference. In summary, for detection of tumor outgrowths, 3D microscopy has almost twice the sensitivity of breadloaf microscopy, particularly in the situation of aggressive/infiltrative carcinomas.

  2. Optical 3D shape, surface, and material analysis

    NASA Astrophysics Data System (ADS)

    Tiziani, Hans J.

    2001-06-01

    Different techniques are available for macro- and micro- topometry. The methods are basically known but their industrial implementation requires robust measuring systems, where calibration is an important necessity. Different techniques will be presented. New elements such as liquid crystal displays and micromirror devices are available leading to new applications to be discussed. Combinative methods and integration in measuring systems becomes interesting. The state of the art and new developments will be presented. Together with calibration for 3D-shock or vibration analysis an object shape measuring systems will be directly combined with a vibration measuring system.

  3. Volumetric label-free imaging and 3D reconstruction of mammalian cochlea based on two-photon excitation fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Zhang, Xianzeng; Geng, Yang; Ye, Qing; Zhan, Zhenlin; Xie, Shusen

    2013-11-01

    The visualization of the delicate structure and spatial relationship of intracochlear sensory cells has relied on the laborious procedures of tissue excision, fixation, sectioning and staining for light and electron microscopy. Confocal microscopy is advantageous for its high resolution and deep penetration depth, yet disadvantageous due to the necessity of exogenous labeling. In this study, we present the volumetric imaging of rat cochlea without exogenous dyes using a near-infrared femtosecond laser as the excitation mechanism and endogenous two-photon excitation fluorescence (TPEF) as the contrast mechanism. We find that TPEF exhibits strong contrast, allowing cellular and even subcellular resolution imaging of the cochlea, differentiating cell types, visualizing delicate structures and the radial nerve fiber. Our results further demonstrate that 3D reconstruction rendered with z-stacks of optical sections enables better revealment of fine structures and spatial relationships, and easily performed morphometric analysis. The TPEF-based optical biopsy technique provides great potential for new and sensitive diagnostic tools for hearing loss or hearing disorders, especially when combined with fiber-based microendoscopy.

  4. 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.

  5. Polysilane-based 3D waveguides for optical interconnects

    NASA Astrophysics Data System (ADS)

    Ogura, Kouhei; Oka, Takeshi; Watanabe, Emi; Aoi, Kazunori; Tsushima, Hiroshi; Okano, Hiroaki; Suzuki, Shuji; Hiramatsu, Seiki

    2008-02-01

    We have been developing the optical waveguide for the multimode using the photo-bleaching property of polysilane. The refractive index of polysilane can be easily changed by exposing to UV light as photobleaching. Using this property, we can make waveguide with simple processes as spin coating, exposing and annealing. We found that this waveguide has thermal adhesive property with glass substrate. And we applied this feature to fabricate multilayered optical waveguides that have three-dimensional structure and can change the optical light at right angle.

  6. 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.

  7. Three-dimensional optical-resolution photoacoustic microscopy.

    PubMed

    Hu, Song; Maslov, Konstantin; Wang, Lihong V

    2011-05-03

    Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As the mainstays of in vivo three-dimensional (3-D) optical microscopy, single-/multi-photon fluorescence microscopy and optical coherence tomography (OCT) have demonstrated their extraordinary sensitivities to fluorescence and optical scattering contrasts, respectively. However, the optical absorption contrast of biological tissues, which encodes essential physiological/pathological information, has not yet been assessable. The emergence of biomedical photoacoustics has led to a new branch of optical microscopy optical-resolution photoacoustic microscopy (OR-PAM), where the optical irradiation is focused to the diffraction limit to achieve cellular or even subcellular level lateral resolution. As a valuable complement to existing optical microscopy technologies, OR-PAM brings in at least two novelties. First and most importantly, OR-PAM detects optical absorption contrasts with extraordinary sensitivity (i.e., 100%). Combining OR-PAM with fluorescence microscopy or with optical-scattering-based OCT (or with both) provides comprehensive optical properties of biological tissues. Second, OR-PAM encodes optical absorption into acoustic waves, in contrast to the pure optical processes in fluorescence microscopy and OCT, and provides background-free detection. The acoustic detection in OR-PAM mitigates the impacts of optical scattering on signal degradation and naturally eliminates possible interferences (i.e., crosstalks) between excitation and detection, which is a common problem in fluorescence microscopy due to the overlap between the excitation and fluorescence spectra. Unique for optical absorption imaging, OR-PAM has demonstrated broad biomedical applications since its invention, including, but not limited to, neurology, ophthalmology, vascular biology, and dermatology. In this video, we teach the system

  8. Kinect the dots: 3D control of optical tweezers

    NASA Astrophysics Data System (ADS)

    Shaw, Lucy; Preece, Daryl; Rubinsztein-Dunlop, Halina

    2013-07-01

    Holographically generated optical traps confine micron- and sub-micron sized particles close to the center of focused light beams. They also provide a way of trapping multiple particles and moving them in three dimensions. However, in many systems the user interface is not always advantageous or intuitive especially for collaborative work and when depth information is required. We discuss and evaluate a set of multi-beam optical tweezers that utilize off the shelf gaming technology to facilitate user interaction. We use the Microsoft Kinect sensor bar as a way of getting the user input required to generate arbitrary optical force fields and control optically trapped particles. We demonstrate that the system can also be used for dynamic light control.

  9. An Optically-Assisted 3-D Cellular Array Machine

    DTIC Science & Technology

    1993-11-05

    Presented by: Physical Optics Corporation 0 Research & Development Division 20600 Gramercy Place, Suite 103 Torrance, California 90501 Principal...Computer Machine (Constructed Hardware) (Planned Hardware Design) Processing Techniques Digital Only Digital and Analog Analog Processor N/A Celular Neural

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

    PubMed

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

    2014-09-29

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

  11. Super-resolution Imaging of the Cytokinetic Z Ring in Live Bacteria Using Fast 3D-Structured Illumination Microscopy (f3D-SIM)

    PubMed Central

    Liew, Andrew T. F.; Monahan, Leigh G.; Whitchurch, Cynthia B.; Harry, Elizabeth J.

    2014-01-01

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

  12. 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.

  13. 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.

  14. Laminar optical tomography: high-resolution 3D functional imaging of superficial tissues

    NASA Astrophysics Data System (ADS)

    Hillman, Elizabeth M. C.; Devor, Anna; Dunn, Andrew K.; Boas, David A.

    2006-03-01

    Laminar Optical Tomography (LOT) is a new medical imaging modality for high-resolution, depth-resolved, functional imaging of superficial tissue such as rodent cortex, skin and the retina. LOT uses visible laser light to image to depths of >2mm (far deeper than microscopy) and is highly sensitive to absorption and fluorescence contrast, enabling spectroscopic functional information such as hemoglobin oxygenation to be imaged with 100-200 micron resolution. LOT has been used to image the hemodynamic response to stimulus in the somatosensory cortex of rats. The resulting three-dimensional (3D) images through the depth of the cortex can be used to delineate the arterial, capillary and venous responses, revealing new information about the intricacies of the oxygenation and blood flow dynamics related to neuronal activation. Additional applications of LOT are being explored, including the integration of 3D Voltage Sensitive Dye fluorescence imaging. LOT imaging uses a system similar to a confocal microscope, quickly scanning a focused beam of light over the surface of the tissue (~8Hz frame rate). Light is detected from both the focus of the scanning beam, and also at increasing distances from the beam's focus. This scattered light has penetrated more deeply into the tissue, and allows features at different depths to be distinguished. An algorithm that includes photon migration modeling of light scattering converts the raw data into 3D images. The motivation for functional optical imaging will be outlined, the basic principles of LOT imaging will be described, and the latest in-vivo results will be presented.

  15. Programmable Bidirectional Folding of Metallic Thin Films for 3D Chiral Optical Antennas.

    PubMed

    Mao, Yifei; Zheng, Yun; Li, Can; Guo, Lin; Pan, Yini; Zhu, Rui; Xu, Jun; Zhang, Weihua; Wu, Wengang

    2017-03-10

    3D structures with characteristic lengths ranging from nanometer to micrometer scale often exhibit extraordinary optical properties, and have been becoming an extensively explored field for building new generation nanophotonic devices. Albeit a few methods have been developed for fabricating 3D optical structures, constructing 3D structures with nanometer accuracy, diversified materials, and perfect morphology is an extremely challenging task. This study presents a general 3D nanofabrication technique, the focused ion beam stress induced deformation process, which allows a programmable and accurate bidirectional folding (-70°-+90°) of various metal and dielectric thin films. Using this method, 3D helical optical antennas with different handedness, improved surface smoothness, and tunable geometries are fabricated, and the strong optical rotation effects of single helical antennas are demonstrated.

  16. Streaming level set algorithm for 3D segmentation of confocal microscopy images.

    PubMed

    Gouaillard, Alexandre; Mosaliganti, Kishore; Gelas, Arnaud; Souhait, Lydie; Obholzer, Nikolaus; Megason, Sean

    2009-01-01

    We present a high performance variant of the popular geodesic active contours which are used for splitting cell clusters in microscopy images. Previously, we implemented a linear pipelined version that incorporates as many cues as possible into developing a suitable level-set speed function so that an evolving contour exactly segments a cell/nuclei blob. We use image gradients, distance maps, multiple channel information and a shape model to drive the evolution. We also developed a dedicated seeding strategy that uses the spatial coherency of the data to generate an over complete set of seeds along with a quality metric which is further used to sort out which seed should be used for a given cell. However, the computational performance of any level-set methodology is quite poor when applied to thousands of 3D data-sets each containing thousands of cells. Those data-sets are common in confocal microscopy. In this work, we explore methods to stream the algorithm in shared memory, multi-core environments. By partitioning the input and output using spatial data structures we insure the spatial coherency needed by our seeding algorithm as well as improve drastically the speed without memory overhead. Our results show speed-ups up to a factor of six.

  17. Immuno- and correlative light microscopy-electron tomography methods for 3D protein localization in yeast.

    PubMed

    Mari, Muriel; Geerts, Willie J C; Reggiori, Fulvio

    2014-10-01

    Compartmentalization of eukaryotic cells is created and maintained through membrane rearrangements that include membrane transport and organelle biogenesis. Three-dimensional reconstructions with nanoscale resolution in combination with protein localization are essential for an accurate molecular dissection of these processes. The yeast Saccharomyces cerevisiae is a key model system for identifying genes and characterizing pathways essential for the organization of cellular ultrastructures. Electron microscopy studies of yeast, however, have been hampered by the presence of a cell wall that obstructs penetration of resins and cryoprotectants, and by the protein dense cytoplasm, which obscures the membrane details. Here we present an immuno-electron tomography (IET) method, which allows the determination of protein distribution patterns on reconstructed organelles from yeast. In addition, we extend this IET approach into a correlative light microscopy-electron tomography procedure where structures positive for a specific protein localized through a fluorescent signal are resolved in 3D. These new investigative tools for yeast will help to advance our understanding of the endomembrane system organization in eukaryotic cells.

  18. Parallel beam optical tomography apparatus for 3D radiation dosimetry

    NASA Astrophysics Data System (ADS)

    Krstajic, Nikola; Doran, Simon J.

    2005-06-01

    Since the discovery of X rays radiotherapy has had the same aim - to deliver a precisely measured dose of radiation to a defined tumour volume with minimal damage to surrounding healthy tissue. Recent developments in radiotherapy such as intensity modulated radiotherapy (IMRT) can generate complex shapes of dose distributions. Until recently it has not been possible to verify that the delivered dose matches the planned dose. However, one often wants to know the real three-dimensional dose distribution. Three-dimensional radiation dosimeters have been developed since the early 1980s. Most chemical formulations involve a radiosensitive species immobilised in space by gelling agent. Magnetic Resonance Imaging (MRI) and optical techniques have been the most successful gel scanning techniques so far. Optical techniques rely on gels changing colour once irradiated. Parallel beam optical tomography has been developed at the University of Surrey since the late 1990s. The apparatus involves light emitting diode light source collimated to a wide (12cm) parallel beam. The beam is attenuated or scattered (depending on the chemical formulation) as it passes through the gel. Focusing optics projects the beam onto a CCD chip. The dosimeter sits on a rotation stage. The tomography scan involves continuously rotating the dosimeter and taking CCD images. Once the dosimeter has been rotated over 180 degrees the images are processed by filtered back projection. The work presented discusses the optics of the apparatus in more detail.

  19. 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

  20. 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.

  1. 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.

  2. Subtractive 3D Printing of Optically Active Diamond Structures

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-05-01

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

  4. Rotating and Precessing Dissipative-Optical-Topological-3D Solitons

    NASA Astrophysics Data System (ADS)

    Veretenov, N. A.; Rosanov, N. N.; Fedorov, S. V.

    2016-10-01

    We predict and study a new type of three-dimensional soliton: asymmetric rotating and precessing stable topological-dissipative-optical localized structures in homogeneous media with saturable amplification and absorption. The crucial factor determining their dynamics is the ratio of the diffusion coefficients characterizing the frequency dispersion and angular selectivity (dichroism) of the scheme. These vortex solitons exist and are stable for overcritical values of the selectivity coefficients and can be realized in lasers of large sizes with saturable absorption.

  5. 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

  6. 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

  7. Rapid Prototyping across the Spectrum: RF to Optical 3D Electromagnetic Structures

    DTIC Science & Technology

    2015-11-17

    fabricated using 3D printer . The fill factor decreases radially outwards and the voids are visible in the unit cells as you approach the periphery of the...with thin walls) [29]. Figure 6: Examples of lenses fabricated with AM (a) GRIN lens fabricated using 3D printer . The fill factor decreases...AFRL-RW-EG-TP-2015-002 Rapid Prototyping across the Spectrum: RF to Optical 3D Electromagnetic Structures Jeffery W. Allen Monica S. Allen Brett

  8. High-Resolution Solid Modeling of Biological Samples Imaged with 3D Fluorescence Microscopy

    PubMed Central

    Ferko, Michael C.; Patterson, Brian W.; Butler, Peter J.

    2011-01-01

    Optical-sectioning, digital fluorescence microscopy provides images representing temporally- and spatially-resolved molecular-scale details of the substructures of living cells. To render such images into solid models for further computational analyses, we have developed an integrated system of image acquisition, processing, and rendering, which includes a new empirical technique to correct for axial distortions inherent in fluorescence microscopy due to refractive index mismatches between microscope objective immersion medium, coverslip glass, and water. This system takes advantage of the capabilities of ultra-high numerical aperture objectives (e.g. total internal reflection fluorescence microscopy) and enables faithful three-dimensional rendering of living cells into solid models amenable to further computational analysis. An example of solid modeling of bovine aortic endothelial cells and their nuclei is presented. Since many cellular level events are temporally and spatially confined, such integrated image acquisition, processing, rendering, and computational analysis, will enable, in silico, the generation of new computational models for cell mechanics and signaling. PMID:16758474

  9. 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.

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

    PubMed

    Guo, Fei; Jiang, Wen

    2014-01-01

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

  11. 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

  12. 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.

  13. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

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

    PubMed Central

    Guo, Fei; Jiang, Wen

    2014-01-01

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

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

    SciTech Connect

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

    2016-01-15

    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.

  17. 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.

  18. 3D Printing Optical Engine for Controlling Material Microstructure

    NASA Astrophysics Data System (ADS)

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

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

  19. Model-based segmentation and quantification of subcellular structures in 2D and 3D fluorescent microscopy images

    NASA Astrophysics Data System (ADS)

    Wörz, Stefan; Heinzer, Stephan; Weiss, Matthias; Rohr, Karl

    2008-03-01

    We introduce a model-based approach for segmenting and quantifying GFP-tagged subcellular structures of the Golgi apparatus in 2D and 3D microscopy images. The approach is based on 2D and 3D intensity models, which are directly fitted to an image within 2D circular or 3D spherical regions-of-interest (ROIs). We also propose automatic approaches for the detection of candidates, for the initialization of the model parameters, and for adapting the size of the ROI used for model fitting. Based on the fitting results, we determine statistical information about the spatial distribution and the total amount of intensity (fluorescence) of the subcellular structures. We demonstrate the applicability of our new approach based on 2D and 3D microscopy images.

  20. Optical rotation compensation for a holographic 3D display with a 360 degree horizontal viewing zone.

    PubMed

    Sando, Yusuke; Barada, Daisuke; Yatagai, Toyohiko

    2016-10-20

    A method for a continuous optical rotation compensation in a time-division-based holographic three-dimensional (3D) display with a rotating mirror is presented. Since the coordinate system of wavefronts after the mirror reflection rotates about the optical axis along with the rotation angle, compensation or cancellation is absolutely necessary to fix the reconstructed 3D object. In this study, we address this problem by introducing an optical image rotator based on a right-angle prism that rotates synchronously with the rotating mirror. The optical and continuous compensation reduces the occurrence of duplicate images, which leads to the improvement of the quality of reconstructed images. The effect of the optical rotation compensation is experimentally verified and a demonstration of holographic 3D display with the optical rotation compensation is presented.

  1. 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.

  2. 3D objects enlargement technique using an optical system and multiple SLMs for electronic holography.

    PubMed

    Yamamoto, Kenji; Ichihashi, Yasuyuki; Senoh, Takanori; Oi, Ryutaro; Kurita, Taiichiro

    2012-09-10

    One problem in electronic holography, which is caused by the display performance of spatial light modulators (SLM), is that the size of reconstructed 3D objects is small. Although methods for increasing the size using multiple SLMs have been considered, they typically had the problem that some parts of 3D objects were missing as a result of the gap between adjacent SLMs or 3D objects lost the vertical parallax. This paper proposes a method of resolving this problem by locating an optical system containing a lens array and other components in front of multiple SLMs. We used an optical system and 9 SLMs to construct a device equivalent to an SLM with approximately 74,600,000 pixels and used this to reconstruct 3D objects in both the horizontal and vertical parallax with an image size of 63 mm without losing any part of 3D objects.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2005-08-01

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

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

    PubMed Central

    2011-01-01

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

  9. 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

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

    PubMed

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

    2012-07-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2005-06-01

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

  12. Cryogenic optical localization provides 3D protein structure data with Angstrom resolution.

    PubMed

    Weisenburger, Siegfried; Boening, Daniel; Schomburg, Benjamin; Giller, Karin; Becker, Stefan; Griesinger, Christian; Sandoghdar, Vahid

    2017-02-01

    We introduce Cryogenic Optical Localization in 3D (COLD), a method to localize multiple fluorescent sites within a single small protein with Angstrom resolution. We demonstrate COLD by determining the conformational state of the cytosolic Per-ARNT-Sim domain from the histidine kinase CitA of Geobacillus thermodenitrificans and resolving the four biotin sites of streptavidin. COLD provides quantitative 3D information about small- to medium-sized biomolecules on the Angstrom scale and complements other techniques in structural biology.

  13. Design of extended viewing zone at autostereoscopic 3D display based on diffusing optical element

    NASA Astrophysics Data System (ADS)

    Kim, Min Chang; Hwang, Yong Seok; Hong, Suk-Pyo; Kim, Eun Soo

    2012-03-01

    In this paper, to realize a non-glasses type 3D display as next step from the current glasses-typed 3D display, it is suggested that a viewing zone is designed for the 3D display using DOE (Diffusing Optical Element). Viewing zone of proposed method is larger than that of the current parallax barrier method or lenticular method. Through proposed method, it is shown to enable the expansion and adjustment of the area of viewing zone according to viewing distance.

  14. Optimized data processing for an optical 3D sensor based on flying triangulation

    NASA Astrophysics Data System (ADS)

    Ettl, Svenja; Arold, Oliver; Häusler, Gerd; Gurov, Igor; Volkov, Mikhail

    2013-05-01

    We present data processing methods for an optical 3D sensor based on the measurement principle "Flying Triangulation". The principle enables a motion-robust acquisition of the 3D shape of even complex objects: A hand-held sensor is freely guided around the object while real-time feedback of the measurement progress is delivered during the captioning. Although of high precision, the resulting 3D data usually may exhibit some weaknesses: e.g. outliers might be present and the data size might be too large. We describe the measurement principle and the data processing and conclude with measurement results.

  15. Liquid crystal lens array for 3D microscopy and endoscope application

    NASA Astrophysics Data System (ADS)

    Huang, Yi-Pai; Hsieh, Po-Yuan; Hassanfiroozi, Amir; Chu, Chao-Yu; Hsuan, Yun; Martinez, Manuel; Javidi, Bahram

    2016-06-01

    In this paper, we demonstrate two liquid crystal (LC) lens array devices for 3D microscope and 3D endoscope applications respectively. Compared with the previous 3D biomedical system, the proposed LC lens arrays are not only switchable between 2D and 3D modes, but also are able to adjust focus in both modes. The multi-function liquid crystal lens (MFLC-lens) array with dual layer electrode has diameter 1.42 mm, which is much smaller than the conventional 3D endoscope with double fixed lenses. The hexagonal liquid crystal micro-lens array (HLC-MLA) instead of fixed micro-lens array in 3D light field microscope can extend the effective depth of field from 60 um to 780 um. To achieve the LC lens arrays, a high-resistance layer needs to be coated on the electrodes to generate an ideal gradient electric-field distribution, which can induce a lens-like form of LC molecules. The parameters and characteristics of high-resistance layer are investigated and discussed with an aim to optimize the performance of liquid crystal lens arrays.

  16. Simultaneous calculation of three optical surfaces in the 3D SMS freeform RXI optic

    NASA Astrophysics Data System (ADS)

    Sorgato, Simone; Chaves, Julio; Mohedano, Rubén.; Hernández, Maikel; Blen, José; Benitez, Pablo; Miñano, Juan C.; Grabovickic, Dejan; Thienpont, Hugo; Duerr, Fabian

    2016-09-01

    The Freeform RXI collimator is a remarkable example of advanced nonimaging device designed with the 3D Simultaneous Multiple Surface (SMS) Method. In the original design, two (the front refracting surface and the back mirror) of the three optical surfaces of the RXI are calculated simultaneously and one (the cavity surrounding the source) is fixed by the designer. As a result, the RXI perfectly couples two input wavefronts (coming from the edges of the extended LED source) with two output wavefronts (defining the output beam). This allows for LED lamps able to produce controlled intensity distributions, which can and have been successfully applied to demanding applications like high- and low-beams for Automotive Lighting. Nevertheless, current trends in this field are moving towards smaller headlamps with more shape constraints driven by car design. We present an improved version of the 3D RXI in which also the cavity surface is computed during the design, so that there are three freeform surfaces calculated simultaneously and an additional degree of freedom for controlling the light emission: now the RXI can perfectly couple three input wavefronts with three output wavefronts. The enhanced control over ray beams allows for improved light homogeneity and better pattern definition.

  17. A Microscopic Optically Tracking Navigation System That Uses High-resolution 3D Computer Graphics.

    PubMed

    Yoshino, Masanori; Saito, Toki; Kin, Taichi; Nakagawa, Daichi; Nakatomi, Hirofumi; Oyama, Hiroshi; Saito, Nobuhito

    2015-01-01

    Three-dimensional (3D) computer graphics (CG) are useful for preoperative planning of neurosurgical operations. However, application of 3D CG to intraoperative navigation is not widespread because existing commercial operative navigation systems do not show 3D CG in sufficient detail. We have developed a microscopic optically tracking navigation system that uses high-resolution 3D CG. This article presents the technical details of our microscopic optically tracking navigation system. Our navigation system consists of three components: the operative microscope, registration, and the image display system. An optical tracker was attached to the microscope to monitor the position and attitude of the microscope in real time; point-pair registration was used to register the operation room coordinate system, and the image coordinate system; and the image display system showed the 3D CG image in the field-of-view of the microscope. Ten neurosurgeons (seven males, two females; mean age 32.9 years) participated in an experiment to assess the accuracy of this system using a phantom model. Accuracy of our system was compared with the commercial system. The 3D CG provided by the navigation system coincided well with the operative scene under the microscope. Target registration error for our system was 2.9 ± 1.9 mm. Our navigation system provides a clear image of the operation position and the surrounding structures. Systems like this may reduce intraoperative complications.

  18. 3D printing optical watermark algorithms based on the combination of DWT and Fresnel transformation

    NASA Astrophysics Data System (ADS)

    Hu, Qi; Duan, Jin; Zhai, Di; Wang, LiNing

    2016-10-01

    With the continuous development of industrialization, 3D printing technology steps into individuals' lives gradually, however, the consequential security issue has become the urgent problem which is imminent. This paper proposes the 3D printing optical watermark algorithms based on the combination of DWT and Fresnel transformation and utilizes authorized key to restrict 3D model printing's permissions. Firstly, algorithms put 3D model into affine transform, and take the distance from the center of gravity to the vertex of 3D object in order to generate a one-dimensional discrete signal; then make this signal into wavelet transform and put the transformed coefficient into Fresnel transformation. Use math model to embed watermark information into it and finally generate 3D digital model with watermarking. This paper adopts VC++.NET and DIRECTX 9.0 SDK for combined developing and testing, and the results show that in fixed affine space, achieve the robustness in translation, revolving and proportion transforms of 3D model and better watermark-invisibility. The security and authorization of 3D model have been protected effectively.

  19. Structured light imaging system for structural and optical characterization of 3D tissue-simulating phantoms

    NASA Astrophysics Data System (ADS)

    Liu, Songde; Smith, Zach; Xu, Ronald X.

    2016-10-01

    There is a pressing need for a phantom standard to calibrate medical optical devices. However, 3D printing of tissue-simulating phantom standard is challenged by lacking of appropriate methods to characterize and reproduce surface topography and optical properties accurately. We have developed a structured light imaging system to characterize surface topography and optical properties (absorption coefficient and reduced scattering coefficient) of 3D tissue-simulating phantoms. The system consisted of a hyperspectral light source, a digital light projector (DLP), a CMOS camera, two polarizers, a rotational stage, a translation stage, a motion controller, and a personal computer. Tissue-simulating phantoms with different structural and optical properties were characterized by the proposed imaging system and validated by a standard integrating sphere system. The experimental results showed that the proposed system was able to achieve pixel-level optical properties with a percentage error of less than 11% for absorption coefficient and less than 7% for reduced scattering coefficient for phantoms without surface curvature. In the meanwhile, 3D topographic profile of the phantom can be effectively reconstructed with an accuracy of less than 1% deviation error. Our study demonstrated that the proposed structured light imaging system has the potential to characterize structural profile and optical properties of 3D tissue-simulating phantoms.

  20. 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

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

    NASA Astrophysics Data System (ADS)

    Bezrukova, Alexandra G.

    2005-04-01

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

  2. 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

  3. Design, Simulation and Optimisation of a Fibre-optic 3D Accelerometer

    NASA Astrophysics Data System (ADS)

    Yang, Zhen; Fang, Xiao-Yong; Zhou, Yan; Li, Ya-lin; Yuan, Jie; Cao, Mao-Sheng

    2013-07-01

    Using an inertia pendulum comprised of two prisms, flexible beams and an elastic flake, we present a novel fibre-optic 3D accelerometer design. The total reverse reflection of the cube-corner prism and the spectroscopic property of an orthogonal holographic grating enable the measurement of the two transverse components of the 3D acceleration simultaneously, while the longitudinal component can be determined from the elastic deformation of the flake. Due to optical interferometry, this sensor may provide a wider range, higher sensitivity and better resolving power than other accelerometers. Moreover, we use finite element analysis to study the performance and to optimise the structural design of the sensor.

  4. A virtually imaged defocused array (VIDA) for high-speed 3D microscopy.

    PubMed

    Schonbrun, Ethan; Di Caprio, Giuseppe

    2016-10-01

    We report a method to capture a multifocus image stack based on recording multiple reflections generated by imaging through a custom etalon. The focus stack is collected in a single camera exposure and consequently the information needed for 3D reconstruction is recorded in the camera integration time, which is only 100 µs. We have used the VIDA microscope to temporally resolve the multi-lobed 3D morphology of neutrophil nuclei as they rotate and deform through a microfluidic constriction. In addition, we have constructed a 3D imaging flow cytometer and quantified the nuclear morphology of nearly a thousand white blood cells flowing at a velocity of 3 mm per second. The VIDA microscope is compact and simple to construct, intrinsically achromatic, and the field-of-view and stack number can be easily reconfigured without redesigning diffraction gratings and prisms.

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

  6. 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.

  7. 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

  8. Atomic force microscopy combined with optical microscopy for cells investigation.

    PubMed

    Cascione, Mariafrancesca; de Matteis, Valeria; Rinaldi, Rosaria; Leporatti, Stefano

    2017-01-01

    This review reports on the combined use of the atomic force microscopy (AFM) and several type of optical/fluorescence/laser scanning microscopy for investigating cells. It is shown that the hybrid systems of AFM with optical-derived microscopies enable to study in detail cell surface properties (such as topography), their mechanical properties (e.g., Young's modulus) mechanotransduction phenomena and allow to gain insight into biological-related pathways and mechanisms in the complex nanoworld of cells. Microsc. Res. Tech. 80:109-123, 2017. © 2016 Wiley Periodicals, Inc.

  9. The study of plant tissue by optical coherent microscopy method

    NASA Astrophysics Data System (ADS)

    Chirskaya, V. V.; Margaryants, N. B.; Zhukova, E. V.

    2016-08-01

    The article presents the results of application of the optical coherent microscopy technique using a high-resolution automatic Linnik interference microscope to study the structure of plant tissues exemplified by surface periderm layers of a tuberous nightshade (solánum tuberosum) bulb. The results of 3D visualization of the structure of the sample under examination are provided. Scanning depth was 32 µm, with axial and lateral resolution of the device 1 µm.

  10. Fluorescent stereo microscopy for 3D surface profilometry and deformation mapping.

    PubMed

    Hu, Zhenxing; Luo, Huiyang; Du, Yingjie; Lu, Hongbing

    2013-05-20

    Recently, mechanobiology has received increased attention. For investigation of biofilm and cellular tissue, measurements of the surface topography and deformation in real-time are a pre-requisite for understanding the growth mechanisms. In this paper, a novel three-dimensional (3D) fluorescent microscopic method for surface profilometry and deformation measurements is developed. In this technique a pair of cameras are connected to a binocular fluorescent microscope to acquire micrographs from two different viewing angles of a sample surface doped or sprayed with fluorescent microparticles. Digital image correlation technique is used to search for matching points in the pairing fluorescence micrographs. After calibration of the system, the 3D surface topography is reconstructed from the pair of planar images. When the deformed surface topography is compared with undeformed topography using fluorescent microparticles for movement tracking of individual material points, the full field deformation of the surface is determined. The technique is demonstrated on topography measurement of a biofilm, and also on surface deformation measurement of the biofilm during growth. The use of 3D imaging of the fluorescent microparticles eliminates the formation of bright parts in an image caused by specular reflections. The technique is appropriate for non-contact, full-field and real-time 3D surface profilometry and deformation measurements of materials and structures at the microscale.

  11. 3D mapping of elastic modulus using shear wave optical micro-elastography

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

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

  12. 3D mapping of elastic modulus using shear wave optical micro-elastography

    PubMed Central

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

    2016-01-01

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

  13. Monocular accommodation condition in 3D display types through geometrical optics

    NASA Astrophysics Data System (ADS)

    Kim, Sung-Kyu; Kim, Dong-Wook; Park, Min-Chul; Son, Jung-Young

    2007-09-01

    Eye fatigue or strain phenomenon in 3D display environment is a significant problem for 3D display commercialization. The 3D display systems like eyeglasses type stereoscopic or auto-stereoscopic multiview, Super Multi-View (SMV), and Multi-Focus (MF) displays are considered for detail calculation about satisfaction level of monocular accommodation by geometrical optics calculation means. A lens with fixed focal length is used for experimental verification about numerical calculation of monocular defocus effect caused by accommodation at three different depths. And the simulation and experiment results consistently show relatively high level satisfaction about monocular accommodation at MF display condition. Additionally, possibility of monocular depth perception, 3D effect, at monocular MF display is discussed.

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

    PubMed

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

    2014-12-02

    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.

  17. 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

  18. Innovative simultaneous confocal full-field 3D surface profilometry for in situ automatic optical inspection (AOI)

    NASA Astrophysics Data System (ADS)

    Chen, Liang-Chia; Chang, Yi-Wei

    2010-06-01

    Rapid acquisition of surface 3D contour information using optical detection has attracted tremendous interest in the field of automatic optical inspection (AOI) and how to avoid or minimize environmental vibration or disturbance has become a critical issue in in situ inspection. Owing to its high longitudinal measurability and excellent vertical resolution, optical confocal microscopy has become extremely important for surface profilometry. This study presents a novel simultaneous confocal full-field 3D surface profilometer using structured fringe projection. The developed confocal optical system is capable of acquiring multiple images at various object depths to perform surface 3D reconstruction by a single image shot without the need for time-consuming vertical scanning. In this method, four conjugate image-sensing modules are configured at four different designated focusing positions, which are controlled by a specially designed beam-splitting optical module. A focal-depth response (FDR) curve can be established by fitting the four focus measurements obtained from these designated positions to achieve simultaneous confocal vertical scanning. In addition, using the principle of optical grating projection, a structured fringe pattern is generated for lateral scanning to enhance the spatial measurement resolution. To examine the performance of the developed system, an accurate step-height target and some industrial micro semiconductor components were measured. The results show that the depth measurement resolution can reach up to 0.1 µm and the maximum measurement error is within 1.5% of the overall range, indicating both accuracy and repeatability of the proposed confocal measurement approach.

  19. Disposable optics for microscopy diagnostics.

    PubMed

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

    2015-11-20

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

  20. 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

  1. 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

  2. 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.

  3. 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

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

    PubMed

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

    2005-06-01

    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 60 nm 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.

  5. High resolution 3D dosimetry for microbeam radiation therapy using optical CT

    NASA Astrophysics Data System (ADS)

    McErlean, C.; Bräuer-Krisch, E.; Adamovics, J.; Leach, M. O.; Doran, S. J.

    2015-01-01

    Optical Computed Tomography (CT) is a promising technique for dosimetry of Microbeam Radiation Therapy (MRT), providing high resolution 3D dose maps. Here different MRT irradiation geometries are visualised showing the potential of Optical CT as a tool for future MRT trials. The Peak-to-Valley dose ratio (PVDR) is calculated to be 7 at a depth of 3mm in the radiochromic dosimeter PRESAGE®. This is significantly lower than predicted values and possible reasons for this are discussed.

  6. Visualization of Mesenchymal Stromal Cells in 2Dand 3D-Cultures by Scanning Electron Microscopy with Lanthanide Contrasting.

    PubMed

    Novikov, I A; Vakhrushev, I V; Antonov, E N; Yarygin, K N; Subbot, A M

    2017-02-01

    Mesenchymal stromal cells from deciduous teeth in 2D- and 3D-cultures on culture plastic, silicate glass, porous polystyrene, and experimental polylactoglycolide matrices were visualized by scanning electron microscopy with lanthanide contrasting. Supravital staining of cell cultures with a lanthanide-based dye (neodymium chloride) preserved normal cell morphology and allowed assessment of the matrix properties of the carriers. The developed approach can be used for the development of biomaterials for tissue engineering.

  7. 3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy

    PubMed Central

    Eum, Juneyong; Kwak, Jina; Kim, Hee Joung; Ki, Seoyoung; Lee, Kooyeon; Raslan, Ahmed A.; Park, Ok Kyu; Chowdhury, Md Ashraf Uddin; Her, Song; Kee, Yun; Kwon, Seung-Hae; Hwang, Byung Joon

    2016-01-01

    Environmental contamination by trinitrotoluene is of global concern due to its widespread use in military ordnance and commercial explosives. Despite known long-term persistence in groundwater and soil, the toxicological profile of trinitrotoluene and other explosive wastes have not been systematically measured using in vivo biological assays. Zebrafish embryos are ideal model vertebrates for high-throughput toxicity screening and live in vivo imaging due to their small size and transparency during embryogenesis. Here, we used Single Plane Illumination Microscopy (SPIM)/light sheet microscopy to assess the developmental toxicity of explosive-contaminated water in zebrafish embryos and report 2,4,6-trinitrotoluene-associated developmental abnormalities, including defects in heart formation and circulation, in 3D. Levels of apoptotic cell death were higher in the actively developing tissues of trinitrotoluene-treated embryos than controls. Live 3D imaging of heart tube development at cellular resolution by light-sheet microscopy revealed trinitrotoluene-associated cardiac toxicity, including hypoplastic heart chamber formation and cardiac looping defects, while the real time PCR (polymerase chain reaction) quantitatively measured the molecular changes in the heart and blood development supporting the developmental defects at the molecular level. Identification of cellular toxicity in zebrafish using the state-of-the-art 3D imaging system could form the basis of a sensitive biosensor for environmental contaminants and be further valued by combining it with molecular analysis. PMID:27869673

  8. 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.

  9. 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

  10. 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.

  11. Alignment of 3-D optical coherence tomography scans to correct eye movement using a particle filtering.

    PubMed

    Xu, Juan; Ishikawa, Hiroshi; Wollstein, Gadi; Kagemann, Larry; Schuman, Joel S

    2012-07-01

    Eye movement artifacts occurring during 3-D optical coherence tomography (OCT) scanning is a well-recognized problem that may adversely affect image analysis and interpretation. A particle filtering algorithm is presented in this paper to correct motion in a 3-D dataset by considering eye movement as a target tracking problem in a dynamic system. The proposed particle filtering algorithm is an independent 3-D alignment approach, which does not rely on any reference image. 3-D OCT data is considered as a dynamic system, while the location of each A-scan is represented by the state space. A particle set is used to approximate the probability density of the state in the dynamic system. The state of the system is updated frame by frame to detect A-scan movement. The proposed method was applied on both simulated data for objective evaluation and experimental data for subjective evaluation. The sensitivity and specificity of the x-movement detection were 98.85% and 99.43%, respectively, in the simulated data. For the experimental data (74 3-D OCT images), all the images were improved after z-alignment, while 81.1% images were improved after x-alignment. The proposed algorithm is an efficient way to align 3-D OCT volume data and correct the eye movement without using references.

  12. Investigation of inclined dual-fiber optical tweezers for 3D manipulation and force sensing.

    PubMed

    Liu, Yuxiang; Yu, Miao

    2009-08-03

    Optical tweezers provide a versatile tool in biological and physical researches. Optical tweezers based on optical fibers are more flexible and ready to be integrated when compared with those based on microscope objectives. In this paper, the three-dimensional (3D) trapping ability of an inclined dual-fiber optical tweezers is demonstrated. The trapping efficiency with respect to displacement is experimentally calibrated along two dimensions. The system is studied numerically using a modified ray-optics model. The spring constants obtained in the experiment are predicted by simulations. It is found both experimentally and numerically that there is a critical value for the fiber inclination angle to retain the 3D trapping ability. The inclined dual-fiber optical tweezers are demonstrated to be more robust to z-axis misalignment than the counter-propagating fiber optical tweezers, which is a special case of th former when the fiber inclination angle is 90 masculine. This inclined dual-fiber optical tweezers can serve as both a manipulator and a force sensor in integrated systems, such as microfluidic systems and lab-on-a-chip systems.

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

    PubMed

    Hua, Hong; Javidi, Bahram

    2014-06-02

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

  14. Optically coupled 3D common memory with GaAs on Si structure

    NASA Astrophysics Data System (ADS)

    Hirose, M.; Takata, H.; Koyanagi, M.

    1991-02-01

    An ultra fast data transfer speed is demonstrated for a novel three-dimensional (3D) Static Random Access Memory (SRAM) consisting of multilayer silicon LSI chips on which GaAs LEDs and photodetectors are monolithically integrated for vertical optical interconnections. A unique feature of this system is the capability of parallel data transfer from one memory layer to the upper and lower memory layers by the optical interconnections. The results of static and dynamic simulations of the optically coupled 3D common memory have indicated that a block of 512 bits data can be transferred through four memory layers within 16 nsec. This is an equivalent data transfer speed of 128 Gbits/sec/layer.

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

    PubMed

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

    2013-12-21

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-12-01

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

  17. 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.

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

    PubMed Central

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

    2016-01-01

    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

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

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

    PubMed

    Bruns, T; Schickinger, S; Schneckenburger, H

    2015-10-01

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

  1. Tip/tilt-compensated through-focus scanning optical microscopy

    NASA Astrophysics Data System (ADS)

    Lee, Jun Ho; Park, Jun Hyung; Jeong, Dohwan; Shin, Eun Ji; Park, Chris

    2016-11-01

    Through-Focus Optical Microscopy (TSOM), with nanometer scale lateral and vertical sensitivity matching those of scanning electron microscopy, has been demonstrated to be utilized for 3D inspection and metrology. There have been sensitivity and instability issues in acquiring through-focus images because TSOM 3D information is indirectly extracted by differentiating a target TSOM image from reference TSOM images. This paper first reports on the optical axis instability that occurs during the scanning process of TSOM when implemented in an existing patterned wafer inspection tool by moving the wafer plane; this is followed by quantitative confirmation of the optical/mechanical instability using a new TSOM tool on an optical bench with a Shack-Hartmann wavefront sensor and a tip/tilt sensor. Then, this paper proposes two tip/tilt compensated TSOM optical acquisition methods that can be applied with adaptive optics. The first method simply adopts a tip/tilt mirror with a quad cell in a simple closed loop, while the second method adopts a highorder deformable mirror with a Shack-Hartmann sensor. The second method is able to correct high-order residual aberrations as well as to perform through-focus scanning without z-axis movement, while the first method is easier to implement in pre-existing wafer inspection systems with only minor modification.

  2. Remote artificial eyes using micro-optical circuit for long-distance 3D imaging perception.

    PubMed

    Thammawongsa, Nopparat; Yupapin, Preecha P

    2016-01-01

    A small-scale optical device incorporated with an optical nano-antenna is designed to operate as the remote artificial eye using a tiny conjugate mirror. A basic device known as a conjugate mirror can be formed using the artificial eye device, the partially reflected light intensities from input source are interfered and the 3D whispering gallery modes formed within the ring centers, which can be modulated and propagated to the object. The image pixel is obtained at the center ring and linked with the optic nerve in the remote area via the nano-antenna, which is useful for blind people.

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

    NASA Astrophysics Data System (ADS)

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

    2012-10-01

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

  4. 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

  5. Advances in automated 3-D image analyses of cell populations imaged by confocal microscopy.

    PubMed

    Ancin, H; Roysam, B; Dufresne, T E; Chestnut, M M; Ridder, G M; Szarowski, D H; Turner, J N

    1996-11-01

    Automated three-dimensional (3-D) image analysis methods are presented for rapid and effective analysis of populations of fluorescently labeled cells or nuclei in thick tissue sections that have been imaged three dimensionally using a confocal microscope. The methods presented here greatly improve upon our earlier work (Roysam et al.:J Microsc 173: 115-126, 1994). The principal advances reported are: algorithms for efficient data pre-processing and adaptive segmentation, effective handling of image anisotrophy, and fast 3-D morphological algorithms for separating overlapping or connected clusters utilizing image gradient information whenever available. A particular feature of this method is its ability to separate densely packed and connected clusters of cell nuclei. Some of the challenges overcome in this work include the efficient and effective handling of imaging noise, anisotrophy, and large variations in image parameters such as intensity, object size, and shape. The method is able to handle significant inter-cell, intra-cell, inter-image, and intra-image variations. Studies indicate that this method is rapid, robust, and adaptable. Examples were presented to illustrate the applicability of this approach to analyzing images of nuclei from densely packed regions in thick sections of rat liver, and brain that were labeled with a fluorescent Schiff reagent.

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

    EPA Science Inventory

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

  7. 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...

  8. 3D printing of tissue-simulating phantoms for calibration of biomedical optical devices

    NASA Astrophysics Data System (ADS)

    Zhao, Zuhua; Zhou, Ximing; Shen, Shuwei; Liu, Guangli; Yuan, Li; Meng, Yuquan; Lv, Xiang; Shao, Pengfei; Dong, Erbao; Xu, Ronald X.

    2016-10-01

    Clinical utility of many biomedical optical devices is limited by the lack of effective and traceable calibration methods. Optical phantoms that simulate biological tissues used for optical device calibration have been explored. However, these phantoms can hardly simulate both structural and optical properties of multi-layered biological tissue. To address this limitation, we develop a 3D printing production line that integrates spin coating, light-cured 3D printing and Fused Deposition Modeling (FDM) for freeform fabrication of optical phantoms with mechanical and optical heterogeneities. With the gel wax Polydimethylsiloxane (PDMS), and colorless light-curable ink as matrix materials, titanium dioxide (TiO2) powder as the scattering ingredient, graphite powder and black carbon as the absorption ingredient, a multilayer phantom with high-precision is fabricated. The absorption and scattering coefficients of each layer are measured by a double integrating sphere system. The results demonstrate that the system has the potential to fabricate reliable tissue-simulating phantoms to calibrate optical imaging devices.

  9. Mapping 3D fiber orientation in tissue using dual-angle optical polarization tractography

    PubMed Central

    Wang, Y.; Ravanfar, M.; Zhang, K.; Duan, D.; Yao, G.

    2016-01-01

    Optical polarization tractography (OPT) has recently been applied to map fiber organization in the heart, skeletal muscle, and arterial vessel wall with high resolution. The fiber orientation measured in OPT represents the 2D projected fiber angle in a plane that is perpendicular to the incident light. We report here a dual-angle extension of the OPT technology to measure the actual 3D fiber orientation in tissue. This method was first verified by imaging the murine extensor digitorum muscle placed at various known orientations in space. The accuracy of the method was further studied by analyzing the 3D fiber orientation of the mouse tibialis anterior muscle. Finally we showed that dual-angle OPT successfully revealed the unique 3D “arcade” fiber structure in the bovine articular cartilage. PMID:27867698

  10. Mapping 3D fiber orientation in tissue using dual-angle optical polarization tractography.

    PubMed

    Wang, Y; Ravanfar, M; Zhang, K; Duan, D; Yao, G

    2016-10-01

    Optical polarization tractography (OPT) has recently been applied to map fiber organization in the heart, skeletal muscle, and arterial vessel wall with high resolution. The fiber orientation measured in OPT represents the 2D projected fiber angle in a plane that is perpendicular to the incident light. We report here a dual-angle extension of the OPT technology to measure the actual 3D fiber orientation in tissue. This method was first verified by imaging the murine extensor digitorum muscle placed at various known orientations in space. The accuracy of the method was further studied by analyzing the 3D fiber orientation of the mouse tibialis anterior muscle. Finally we showed that dual-angle OPT successfully revealed the unique 3D "arcade" fiber structure in the bovine articular cartilage.

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

    NASA Astrophysics Data System (ADS)

    Ettl, Svenja

    2015-04-01

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

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

    PubMed

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

    2015-04-01

    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.

  13. 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.

  14. 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.

  15. 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

  16. 3D myofibril imaging in live cardiomyocytes via hybrid SHG-TPEF microscopy

    NASA Astrophysics Data System (ADS)

    Shao, Yonghong; Liu, Honghai; Ye, Tong; Borg, Tom; Qu, Junle; Peng, Xiang; Niu, Hanben; Gao, Bruce

    2011-03-01

    We developed a hybrid SHG-TPEF polarization imaging system that allowed the excitation beam from an fs Ti:Sappire laser being bi-directionally raster scanned across the focal plane using a pair of orthogonal galvanometers. To implement high-speed scanning, the turning regions of the triangular waves were smoothed by a custom-designed waveform. The SHG and TPEF signals from samples were recorded by two PMTs in the forward and backward direction. Using this imaging system, we obtained 3D images of the sarcomere structure via SHG and DiO-stained lipid membrane via TPEF in live cardiomyocytes isolated from neonatal and adult rats. The results demonstrated the potential applications of SHG and TPEF in the research of myofibrillogensis.

  17. 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.

  18. 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.

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

    PubMed

    Romero-Brey, Inés; Bartenschlager, Ralf

    2015-12-03

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

  20. 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

  1. Macro optical projection tomography for large scale 3D imaging of plant structures and gene activity.

    PubMed

    Lee, Karen J I; Calder, Grant M; Hindle, Christopher R; Newman, Jacob L; Robinson, Simon N; Avondo, Jerome J H Y; Coen, Enrico S

    2016-12-26

    Optical projection tomography (OPT) is a well-established method for visualising gene activity in plants and animals. However, a limitation of conventional OPT is that the specimen upper size limit precludes its application to larger structures. To address this problem we constructed a macro version called Macro OPT (M-OPT). We apply M-OPT to 3D live imaging of gene activity in growing whole plants and to visualise structural morphology in large optically cleared plant and insect specimens up to 60 mm tall and 45 mm deep. We also show how M-OPT can be used to image gene expression domains in 3D within fixed tissue and to visualise gene activity in 3D in clones of growing young whole Arabidopsis plants. A further application of M-OPT is to visualise plant-insect interactions. Thus M-OPT provides an effective 3D imaging platform that allows the study of gene activity, internal plant structures and plant-insect interactions at a macroscopic scale.

  2. A high-throughput comparative characterization of laser-induced soft tissue damage using 3D digital microscopy.

    PubMed

    Das, Debobrato; Reed, Stephanie; Klokkevold, Perry R; Wu, Benjamin M

    2013-02-01

    3D digital microscopy was used to develop a rapid alternative approach to quantify the effects of specific laser parameters on soft tissue ablation and charring in vitro without the use of conventional tissue processing techniques. Two diode lasers operating at 810 and 980 nm wavelengths were used to ablate three tissue types (bovine liver, turkey breast, and bovine muscle) at varying laser power (0.3, 1.0, and 2.0 W) and velocities (1-50 mm/s). Spectrophotometric analyses were performed on each tissue to determine tissue-specific absorption coefficients and were considered in creating wavelength-dependent energy attenuation models to evaluate minimum heat of tissue ablations. 3D surface contour profiles characterizing tissue damage revealed that ablation depth and tissue charring increased with laser power and decreased with lateral velocity independent of wavelength and tissue type. While bovine liver ablation and charring were statistically higher at 810 than 980 nm (p < 0.05), turkey breast and bovine muscle ablated and charred more at 980 than 810 nm (p < 0.05). Spectrophotometric analysis revealed that bovine liver tissue had a greater tissue-specific absorption coefficient at 810 than 980 nm, while turkey breast and bovine muscle had a larger absorption coefficient at 980 nm (p < 0.05). This rapid 3D microscopic analysis of robot-driven laser ablation yielded highly reproducible data that supported well-defined trends related to laser-tissue interactions and enabled high throughput characterization of many laser-tissue permutations. Since 3D microscopy quantifies entire lesions without altering the tissue specimens, conventional and immunohistologic techniques can be used, if desired, to further interrogate specific sections of the digitized lesions.

  3. 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

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

    NASA Astrophysics Data System (ADS)

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

    2016-12-01

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

  5. 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

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

    PubMed

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

    2013-02-01

    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.

  7. Fibre-optic nonlinear optical microscopy and endoscopy.

    PubMed

    Fu, L; Gu, M

    2007-06-01

    Nonlinear optical microscopy has been an indispensable laboratory tool of high-resolution imaging in thick tissue and live animals. Rapid developments of fibre-optic components in terms of growing functionality and decreasing size provide enormous opportunities for innovations in nonlinear optical microscopy. Fibre-based nonlinear optical endoscopy is the sole instrumentation to permit the cellular imaging within hollow tissue tracts or solid organs that are inaccessible to a conventional optical microscope. This article reviews the current development of fibre-optic nonlinear optical microscopy and endoscopy, which includes crucial technologies for miniaturized nonlinear optical microscopy and their embodiments of endoscopic systems. A particular attention is given to several classes of photonic crystal fibres that have been applied to nonlinear optical microscopy due to their unique properties for ultrashort pulse delivery and signal collection. Furthermore, fibre-optic nonlinear optical imaging systems can be classified into portable microscopes suitable for imaging behaving animals, rigid endoscopes that allow for deep tissue imaging with minimally invasive manners, and flexible endoscopes enabling imaging of internal organs. Fibre-optic nonlinear optical endoscopy is coming of age and a paradigm shift leading to optical microscope tools for early cancer detection and minimally invasive surgery.

  8. Research in Image Understanding as Applied to 3-D Microwave Tomographic Imaging with Near Optical Resolution.

    DTIC Science & Technology

    1986-03-10

    Severe Clutter .... ........ 1I-i III . Optical Implementation of the HopfieldModel .I -? .- . ." Model........................ . . BY...can be employed in future broad-band imaging radar networks capable of providing 3-D projective or . - tomographic images of remote aerospace targets...We expect the results of this effort to tell us how to achieve centimeter resolution on remote aerospace objects cost-effectively using microwave

  9. Rotary-scanning optical resolution photoacoustic microscopy

    NASA Astrophysics Data System (ADS)

    Qi, Weizhi; Xi, Lei

    2016-10-01

    Optical resolution photoacoustic microscopy (ORPAM) is currently one of the fastest evolving photoacoustic imaging modalities. It has a comparable spatial resolution to pure optical microscopic techniques such as epifluorescence microscopy, confocal microscopy, and two-photon microscopy, but also owns a deeper penetration depth. In this paper, we report a rotary-scanning (RS)-ORPAM that utilizes a galvanometer scanner integrated with objective to achieve rotary laser scanning. A 15 MHz cylindrically focused ultrasonic transducer is mounted onto a motorized rotation stage to follow optical scanning traces synchronously. To minimize the loss of signal to noise ratio, the acoustic focus is precisely adjusted to reach confocal with optical focus. Black tapes and carbon fibers are firstly imaged to evaluate the performance of the system, and then in vivo imaging of vasculature networks inside the ears and brains of mice is demonstrated using this system.

  10. EMRinger: side chain–directed model and map validation for 3D cryo-electron microscopy

    DOE PAGES

    Barad, Benjamin A.; Echols, Nathaniel; Wang, Ray Yu-Ruei; ...

    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.

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

    PubMed

    Xu, Y; Wuu, Cheng-Shie; Maryanski, Marek J

    2004-11-01

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

  12. 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

  13. Investigation of Rho Signaling Pathways in 3-D Collagen Matrices with Multidimensional Microscopy and Visualization Techniques

    DTIC Science & Technology

    2008-03-01

    rhotekin. (Rho-GFP; RBD-GFP) -Refining FRET biosensor approach -Refining experimental design and approach to include migration and invasion -Better...Laboratory for Optical and Com- utational Instrumentation, University of Wisconsin at Madison, 271 Animal ciences, 1675 Observatory Drive, Madison, WI 53706...maintained at 37°C ith 10% CO2 until imaged as described in the text. .2 Mammary Tumors ll animal experiments were approved by the institutional nimal use

  14. 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.

  15. 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.

  16. 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.

  17. Wavelength Independent Optical Microscopy and Lithography

    DTIC Science & Technology

    1987-10-31

    Leviatan , Y., J. Appl. Phys. 60, 1577 (1986). 7. Harootunian, A., Near-Field Scanning Optical Microscopy and Raman Microscopy, Cornell University Ph.D...although the approach used may not be valid in the Another potential problem concerns the effect of the near field. More recently, Leviatan 21...Massey, "Microscopy and Pattern Generation With Scanned Evanescent Waves," AppL. Opt. 23, 658 (1984). The authors wish to thank Yehuda Leviatan for 21

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. A surface-based 3-D dendritic spine detection approach from confocal microscopy images.

    PubMed

    Li, Qing; Deng, Zhigang

    2012-03-01

    Determining the relationship between the dendritic spine morphology and its functional properties is a fundamental challenge in neurobiology research. In particular, how to accurately and automatically analyse meaningful structural information from a large microscopy image data set is far away from being resolved. As pointed out in existing literature, one remaining challenge in spine detection and segmentation is how to automatically separate touching spines. In this paper, based on various global and local geometric features of the dendrite structure, we propose a novel approach to detect and segment neuronal spines, in particular, a breaking-down and stitching-up algorithm to accurately separate touching spines. Extensive performance comparisons show that our approach is more accurate and robust than two state-of-the-art spine detection and segmentation algorithms.

  3. 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

  4. 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.

  5. 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.

  6. 3D restoration microscopy improves quantification of enzyme-labeled fluorescence-based single-cell phosphatase activity in plankton.

    PubMed

    Diaz-de-Quijano, Daniel; Palacios, Pilar; Horňák, Karel; Felip, Marisol

    2014-10-01

    The ELF or fluorescence-labeled enzyme activity (FLEA) technique is a culture-independent single-cell tool for assessing plankton enzyme activity in close-to-in situ conditions. We demonstrate that single-cell FLEA quantifications based on two-dimensional (2D) image analysis were biased by up to one order of magnitude relative to deconvolved 3D. This was basically attributed to out-of-focus light, and partially to object size. Nevertheless, if sufficient cells were measured (25-40 cells), biases in individual 2D cell measurements were partially compensated, providing useful and comparable results to deconvolved 3D. We also discuss how much caution should be used when comparing the single-cell enzyme activities of different sized bacterio- and/or phytoplankton populations measured on 2D images. Finally, a novel method based on deconvolved 3D images (wide field restoration microscopy; WFR) was devised to improve the discrimination of similar single-cell enzyme activities, the comparison of enzyme activities between different size cells, the measurement of low fluorescence intensities, the quantification of less numerous species, and the combination of the FLEA technique with other single-cell methods. These improvements in cell enzyme activity measurements will provide a more precise picture of individual species' behavior in nature, which is essential to understand their functional role and evolutionary history.

  7. 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.

  8. Real-time monitoring of quorum sensing in 3D-printed bacterial aggregates using scanning electrochemical microscopy.

    PubMed

    Connell, Jodi L; Kim, Jiyeon; Shear, Jason B; Bard, Allen J; Whiteley, Marvin

    2014-12-23

    Microbes frequently live in nature as small, densely packed aggregates containing ∼10(1)-10(5) cells. These aggregates not only display distinct phenotypes, including resistance to antibiotics, but also, serve as building blocks for larger biofilm communities. Aggregates within these larger communities display nonrandom spatial organization, and recent evidence indicates that this spatial organization is critical for fitness. Studying single aggregates as well as spatially organized aggregates remains challenging because of the technical difficulties associated with manipulating small populations. Micro-3D printing is a lithographic technique capable of creating aggregates in situ by printing protein-based walls around individual cells or small populations. This 3D-printing strategy can organize bacteria in complex arrangements to investigate how spatial and environmental parameters influence social behaviors. Here, we combined micro-3D printing and scanning electrochemical microscopy (SECM) to probe quorum sensing (QS)-mediated communication in the bacterium Pseudomonas aeruginosa. Our results reveal that QS-dependent behaviors are observed within aggregates as small as 500 cells; however, aggregates larger than 2,000 bacteria are required to stimulate QS in neighboring aggregates positioned 8 μm away. These studies provide a powerful system to analyze the impact of spatial organization and aggregate size on microbial behaviors.

  9. Highlighting the impact of aging on type I collagen: label-free investigation using confocal reflectance microscopy and diffuse reflectance spectroscopy in 3D matrix model.

    PubMed

    Guilbert, Marie; Roig, Blandine; Terryn, Christine; Garnotel, Roselyne; Jeannesson, Pierre; Sockalingum, Ganesh D; Manfait, Michel; Perraut, François; Dinten, Jean-Marc; Koenig, Anne; Piot, Olivier

    2016-02-23

    During aging, alterations of extracellular matrix proteins contribute to various pathological phenotypes. Among these alterations, type I collagen cross-linking and associated glycation products accumulation over time detrimentally affects its physico-chemical properties, leading to alterations of tissue biomechanical stability. Here, different-age collagen 3D matrices using non-destructive and label-free biophotonic techniques were analysed to highlight the impact of collagen I aging on 3D constructs, at macroscopic and microscopic levels. Matrices were prepared with collagens extracted from tail tendons of rats (newborns, young and old adults) to be within the physiological aging process. The data of diffuse reflectance spectroscopy reveal that aging leads to an inhibition of fibril assembly and a resulting decrease of gel density. Investigations by confocal reflectance microscopy highlight poor-fibrillar structures in oldest collagen networks most likely related to the glycation products accumulation. Complementarily, an infrared analysis brings out marked spectral variations in the Amide I profile, specific of the peptidic bond conformation and for carbohydrates vibrations as function of collagen-age. Interestingly, we also highlight an unexpected behavior for newborn collagen, exhibiting poorly-organized networks and microscopic features close to the oldest collagen. These results demonstrate that changes in collagen optical properties are relevant for investigating the incidence of aging in 3D matrix models.

  10. 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

  11. Genetically targeted 3D visualisation of Drosophila neurons under Electron Microscopy and X-Ray Microscopy using miniSOG

    PubMed Central

    Ng, Julian; Browning, Alyssa; Lechner, Lorenz; Terada, Masako; Howard, Gillian; Jefferis, Gregory S. X. E.

    2016-01-01

    Large dimension, high-resolution imaging is important for neural circuit visualisation as neurons have both long- and short-range patterns: from axons and dendrites to the numerous synapses at terminal endings. Electron Microscopy (EM) is the favoured approach for synaptic resolution imaging but how such structures can be segmented from high-density images within large volume datasets remains challenging. Fluorescent probes are widely used to localise synapses, identify cell-types and in tracing studies. The equivalent EM approach would benefit visualising such labelled structures from within sub-cellular, cellular, tissue and neuroanatomical contexts. Here we developed genetically-encoded, electron-dense markers using miniSOG. We demonstrate their ability in 1) labelling cellular sub-compartments of genetically-targeted neurons, 2) generating contrast under different EM modalities, and 3) segmenting labelled structures from EM volumes using computer-assisted strategies. We also tested non-destructive X-ray imaging on whole Drosophila brains to evaluate contrast staining. This enabled us to target specific regions for EM volume acquisition. PMID:27958322

  12. A comprehensive evaluation of the PRESAGE/optical-CT 3D dosimetry system

    SciTech Connect

    Sakhalkar, H. S.; Adamovics, J.; Ibbott, G.; Oldham, M.

    2009-01-15

    This work presents extensive investigations to evaluate the robustness (intradosimeter consistency and temporal stability of response), reproducibility, precision, and accuracy of a relatively new 3D dosimetry system comprising a leuco-dye doped plastic 3D dosimeter (PRESAGE) and a commercial optical-CT scanner (OCTOPUS 5x scanner from MGS Research, Inc). Four identical PRESAGE 3D dosimeters were created such that they were compatible with the Radiologic Physics Center (RPC) head-and-neck (H and N) IMRT credentialing phantom. Each dosimeter was irradiated with a rotationally symmetric arrangement of nine identical small fields (1x3 cm{sup 2}) impinging on the flat circular face of the dosimeter. A repetitious sequence of three dose levels (4, 2.88, and 1.28 Gy) was delivered. The rotationally symmetric treatment resulted in a dose distribution with high spatial variation in axial planes but only gradual variation with depth along the long axis of the dosimeter. The significance of this treatment was that it facilitated accurate film dosimetry in the axial plane, for independent verification. Also, it enabled rigorous evaluation of robustness, reproducibility and accuracy of response, at the three dose levels. The OCTOPUS 5x commercial scanner was used for dose readout from the dosimeters at daily time intervals. The use of improved optics and acquisition technique yielded substantially improved noise characteristics (reduced to {approx}2%) than has been achieved previously. Intradosimeter uniformity of radiochromic response was evaluated by calculating a 3D gamma comparison between each dosimeter and axially rotated copies of the same dosimeter. This convenient technique exploits the rotational symmetry of the distribution. All points in the gamma comparison passed a 2% difference, 1 mm distance-to-agreement criteria indicating excellent intradosimeter uniformity even at low dose levels. Postirradiation, the dosimeters were all found to exhibit a slight increase in

  13. 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

  14. 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.

  15. Master-slave interferometry for parallel spectral domain interferometry sensing and versatile 3D optical coherence tomography.

    PubMed

    Podoleanu, Adrian Gh; Bradu, Adrian

    2013-08-12

    Conventional spectral domain interferometry (SDI) methods suffer from the need of data linearization. When applied to optical coherence tomography (OCT), conventional SDI methods are limited in their 3D capability, as they cannot deliver direct en-face cuts. Here we introduce a novel SDI method, which eliminates these disadvantages. We denote this method as Master - Slave Interferometry (MSI), because a signal is acquired by a slave interferometer for an optical path difference (OPD) value determined by a master interferometer. The MSI method radically changes the main building block of an SDI sensor and of a spectral domain OCT set-up. The serially provided signal in conventional technology is replaced by multiple signals, a signal for each OPD point in the object investigated. This opens novel avenues in parallel sensing and in parallelization of signal processing in 3D-OCT, with applications in high- resolution medical imaging and microscopy investigation of biosamples. Eliminating the need of linearization leads to lower cost OCT systems and opens potential avenues in increasing the speed of production of en-face OCT images in comparison with conventional SDI.

  16. 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.

  17. 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

  18. 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.

  19. A new way to characterize autostereoscopic 3D displays using Fourier optics instrument

    NASA Astrophysics Data System (ADS)

    Boher, P.; Leroux, T.; Bignon, T.; Collomb-Patton, V.

    2009-02-01

    Auto-stereoscopic 3D displays offer presently the most attractive solution for entertainment and media consumption. Despite many studies devoted to this type of technology, efficient characterization methods are still missing. We present here an innovative optical method based on high angular resolution viewing angle measurements with Fourier optics instrument. This type of instrument allows measuring the full viewing angle aperture of the display very rapidly and accurately. The system used in the study presents a very high angular resolution below 0.04 degree which is mandatory for this type of characterization. We can predict from the luminance or color viewing angle measurements of the different views of the 3D display what will be seen by an observer at any position in front of the display. Quality criteria are derived both for 3D and standard properties at any observer position and Qualified Stereo Viewing Space (QSVS) is determined. The use of viewing angle measurements at different locations on the display surface during the observer computation gives more realistic estimation of QSVS and ensures its validity for the entire display surface. Optimum viewing position, viewing freedom, color shifts and standard parameters are also quantified. Simulation of the moire issues can be made leading to a better understanding of their origin.

  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. Optical absorption enhancement in 3D nanofibers coated on polymer substrate for photovoltaic devices.

    PubMed

    Kiani, Amirkianoosh; Venkatakrishnan, Krishnan; Tan, Bo

    2015-06-01

    Recent research in the field of photovoltaics has shown that polymer solar cells have great potential to provide low-cost, lightweight and flexible electronic devices to harvest solar energy. In this paper, we propose a new method for the generation of three-dimensional nanofibers coated on polymer substrate induced by femtosecond laser pulses. In this new method, a thin layer of polymer is irradiated by megahertz femtosecond laser pulses under ambient conditions, and a thin fibrous layer is generated on top of the polymer substrate. This method is single step; no additional materials are added, and the layers of the three-dimensional (3D) polymer nanofibrous structures are grown on top of the substrate after laser irradiation. Light spectroscopy results show significant enhancement of light absorption in the generated 3D nanofibrous layers of polymer. Finally, we suggest how to maximize the light trapping and optical absorption of the generated nanofiber cells by optimizing the laser parameters.

  2. Three-Axis Distributed Fiber Optic Strain Measurement in 3D Woven Composite Structures

    NASA Technical Reports Server (NTRS)

    Castellucci, Matt; Klute, Sandra; Lally, Evan M.; Froggatt, Mark E.; Lowry, David

    2013-01-01

    Recent advancements in composite materials technologies have broken further from traditional designs and require advanced instrumentation and analysis capabilities. Success or failure is highly dependent on design analysis and manufacturing processes. By monitoring smart structures throughout manufacturing and service life, residual and operational stresses can be assessed and structural integrity maintained. Composite smart structures can be manufactured by integrating fiber optic sensors into existing composite materials processes such as ply layup, filament winding and three-dimensional weaving. In this work optical fiber was integrated into 3D woven composite parts at a commercial woven products manufacturing facility. The fiber was then used to monitor the structures during a VARTM manufacturing process, and subsequent static and dynamic testing. Low cost telecommunications-grade optical fiber acts as the sensor using a high resolution commercial Optical Frequency Domain Reflectometer (OFDR) system providing distributed strain measurement at spatial resolutions as low as 2mm. Strain measurements using the optical fiber sensors are correlated to resistive strain gage measurements during static structural loading. Keywords: fiber optic, distributed strain sensing, Rayleigh scatter, optical frequency domain reflectometry

  3. Efficient physics-based predictive 3D image modeling and simulation of optical atmospheric refraction phenomena

    NASA Astrophysics Data System (ADS)

    Reinhardt, Colin N.; Hammel, Stephen M.; Tsintikidis, Dimitris

    2016-09-01

    We present some preliminary results and discussion of our ongoing effort to develop a prototype volumetric atmospheric optical refraction simulator which uses 3D nonlinear ray-tracing and state-of-art physics-based rendering techniques. The tool will allow simulation of optical curved-ray propagation through nonlinear refractivity gradient profiles in volumetric atmospheric participating media, and the generation of radiometrically accurate images of the resulting atmospheric refraction phenomena, including inferior and superior mirages, over-the-horizon viewing conditions, looming and sinking, towering and stooping of distant objects. The ability to accurately model and predict atmospheric optical refraction conditions and phenomena is important in both defense and commercial applications. Our nonlinear refractive ray-trace method is currently CPU-parallelized and is well-suited for GPU compute implementation.

  4. Influences of edges and steep slopes in 3D interference and confocal microscopy

    NASA Astrophysics Data System (ADS)

    Xie, Weichang; Hagemeier, Sebastian; Woidt, Carsten; Hillmer, Harmut; Lehmann, Peter

    2016-04-01

    Optical measurement techniques are widely applied in high-resolution contour, topography and roughness measurement. In this context vertical scanning white-light interferometers and confocal microscopes have become mature instruments over the last decades. The accuracy of measurement results is highly related not only to the type and physical properties of the measuring instruments, but also to the measurement object itself. This contribution focuses on measurement effects occurring at edges and height steps using white-light interferometers of different numerical apertures. If the edge is perfectly perpendicular, batwing effects appear at height steps. These batwings show maximum height if the height-to-wavelength-ratio (HWR) is about one forth or three forth, and they disappear if the HWR value is about an integer multiple of one half. The wavelength that is relevant in this context is the effective wavelength, i.e. the center wavelength of the illuminating light multiplied by a correction factor known as the numerical aperture correction. However, in practice the edges are usually not perfectly perpendicular. In this case, the measurement results depend also on the derivative of the surface height function and they may differ from theory and the prediction according to the HWR value. Measurements of such steps show systematical effects depending on the lateral resolution of the instrument. In this context, a Linnik interferometer with a magnification of 100x and NA = 0.9 is used to characterize the three dimensional topography of more or less rectangular calibration specimens and quasi-perpendicular structures produced by the nanoimprint technology. The Linnik interferometer is equipped with LED light sources emitting at different wavelengths, so that the HWR value can be changed. This is possible since the high NA objective lenses show a rather limited depth of focus such that the temporal coherence gating may be replaced by focal gating in this particular

  5. A Marked Poisson Process Driven Latent Shape Model for 3D Segmentation of Reflectance Confocal Microscopy Image Stacks of Human Skin.

    PubMed

    Ghanta, Sindhu; Jordan, Michael I; Kose, Kivanc; Brooks, Dana H; Rajadhyaksha, Milind; Dy, Jennifer G

    2016-10-05

    Segmenting objects of interest from 3D datasets is a common problem encountered in biological data. Small field of view and intrinsic biological variability combined with optically subtle changes of intensity, resolution and low contrast in images make the task of segmentation difficult, especially for microscopy of unstained living or freshly excised thick tissues. Incorporating shape information in addition to the appearance of the object of interest can often help improve segmentation performance. However, shapes of objects in tissue can be highly variable and design of a flexible shape model that encompasses these variations is challenging. To address such complex segmentation problems, we propose a unified probabilistic framework that can incorporate the uncertainty associated with complex shapes, variable appearance and unknown locations. The driving application which inspired the development of this framework is a biologically important segmentation problem: the task of automatically detecting and segmenting the dermal-epidermal junction (DEJ) in 3D reflectance confocal microscopy (RCM) images of human skin. RCM imaging allows noninvasive observation of cellular, nuclear and morphological detail. The DEJ is an important morphological feature as it is where disorder, disease and cancer usually start. Detecting the DEJ is challenging because it is a 2D surface in a 3D volume which has strong but highly variable number of irregularly spaced and variably shaped "peaks and valleys". In addition, RCM imaging resolution, contrast and intensity vary with depth. Thus a prior model needs to incorporate the intrinsic structure while allowing variability in essentially all its parameters. We propose a model which can incorporate objects of interest with complex shapes and variable appearance in an unsupervised setting by utilizing domain knowledge to build appropriate priors of the model. Our novel strategy to model this structure combines a spatial Poisson process with

  6. A Marked Poisson Process Driven Latent Shape Model for 3D Segmentation of Reflectance Confocal Microscopy Image Stacks of Human Skin.

    PubMed

    Ghanta, Sindhu; Jordan, Michael I; Kose, Kivanc; Brooks, Dana H; Rajadhyaksha, Milind; Dy, Jennifer G

    2017-01-01

    Segmenting objects of interest from 3D data sets is a common problem encountered in biological data. Small field of view and intrinsic biological variability combined with optically subtle changes of intensity, resolution, and low contrast in images make the task of segmentation difficult, especially for microscopy of unstained living or freshly excised thick tissues. Incorporating shape information in addition to the appearance of the object of interest can often help improve segmentation performance. However, the shapes of objects in tissue can be highly variable and design of a flexible shape model that encompasses these variations is challenging. To address such complex segmentation problems, we propose a unified probabilistic framework that can incorporate the uncertainty associated with complex shapes, variable appearance, and unknown locations. The driving application that inspired the development of this framework is a biologically important segmentation problem: the task of automatically detecting and segmenting the dermal-epidermal junction (DEJ) in 3D reflectance confocal microscopy (RCM) images of human skin. RCM imaging allows noninvasive observation of cellular, nuclear, and morphological detail. The DEJ is an important morphological feature as it is where disorder, disease, and cancer usually start. Detecting the DEJ is challenging, because it is a 2D surface in a 3D volume which has strong but highly variable number of irregularly spaced and variably shaped "peaks and valleys." In addition, RCM imaging resolution, contrast, and intensity vary with depth. Thus, a prior model needs to incorporate the intrinsic structure while allowing variability in essentially all its parameters. We propose a model which can incorporate objects of interest with complex shapes and variable appearance in an unsupervised setting by utilizing domain knowledge to build appropriate priors of the model. Our novel strategy to model this structure combines a spatial Poisson

  7. 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.

  8. Three-dimensional microscopy by optical scanning holography

    NASA Astrophysics Data System (ADS)

    Poon, Ting-Chung; Doh, Kyu B.; Schilling, Bradley W.; Wu, Ming H.; Shinoda, Kazunori K.; Suzuki, Yoshiji

    1995-05-01

    We first briefly review a new 3D imaging technique called optical scanning holography (OSH). We then discuss the technique's 3D holographic magnification in the context of optical scanning and digital reconstruction. Finally, we demonstrate the 3D imaging capability of OSH by holographically recording two planar objects at different depths and reconstructing the hologram digitally.

  9. Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Yu, Jiun-Yann; Kuo, Chun-Hung; Holland, Daniel B.; Chen, Yenyu; Ouyang, Mingxing; Blake, Geoffrey A.; Zadoyan, Ruben; Guo, Chin-Lin

    2011-11-01

    Optical sectioning provides three-dimensional (3D) information in biological tissues. However, most imaging techniques implemented with optical sectioning are either slow or deleterious to live tissues. Here, we present a simple design for wide-field multiphoton microscopy, which provides optical sectioning at a reasonable frame rate and with a biocompatible laser dosage. The underlying mechanism of optical sectioning is diffuser-based temporal focusing. Axial resolution comparable to confocal microscopy is theoretically derived and experimentally demonstrated. To achieve a reasonable frame rate without increasing the laser power, a low-repetition-rate ultrafast laser amplifier was used in our setup. A frame rate comparable to that of epifluorescence microscopy was demonstrated in the 3D imaging of fluorescent protein expressed in live epithelial cell clusters. In this report, our design displays the potential to be widely used for video-rate live-tissue and embryo imaging with axial resolution comparable to laser scanning microscopy.

  10. 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.

  11. Fiber optic vibration sensor for high-power electric machines realized using 3D printing technology

    NASA Astrophysics Data System (ADS)

    Igrec, Bojan; Bosiljevac, Marko; Sipus, Zvonimir; Babic, Dubravko; Rudan, Smiljko

    2016-03-01

    The objective of this work was to demonstrate a lightweight and inexpensive fiber-optic vibration sensor, built using 3D printing technology, for high-power electric machines and similar applications. The working principle is based on modulating the light intensity using a blade attached to a bendable membrane. The sensor prototype was manufactured using PolyJet Matrix technology with DM 8515 Grey 35 Polymer. The sensor shows linear response, expected bandwidth (< 150 Hz), and from our measurements we estimated the damping ratio for used polymer to be ζ ≍ 0.019. The developed prototype is simple to assemble, adjust, calibrate and repair.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization

    PubMed Central

    Holden, Seamus J.; Pengo, Thomas; Meibom, Karin L.; Fernandez Fernandez, Carmen; Collier, Justine; Manley, Suliana

    2014-01-01

    We created a high-throughput modality of photoactivated localization microscopy (PALM) that enables automated 3D PALM imaging of hundreds of synchronized bacteria during all stages of the cell cycle. We used high-throughput PALM to investigate the nanoscale organization of the bacterial cell division protein FtsZ in live Caulobacter crescentus. We observed that FtsZ predominantly localizes as a patchy midcell band, and only rarely as a continuous ring, supporting a model of “Z-ring” organization whereby FtsZ protofilaments are randomly distributed within the band and interact only weakly. We found evidence for a previously unidentified period of rapid ring contraction in the final stages of the cell cycle. We also found that DNA damage resulted in production of high-density continuous Z-rings, which may obstruct cytokinesis. Our results provide a detailed quantitative picture of in vivo Z-ring organization. PMID:24616530

  19. Super-resolution optical microscopy: multiple choices.

    PubMed

    Huang, Bo

    2010-02-01

    The recent invention of super-resolution optical microscopy enables the visualization of fine features in biological samples with unprecedented clarity. It creates numerous opportunities in biology because vast amount of previously obscured subcellular processes now can be directly observed. Rapid development in this field in the past two years offers many imaging modalities that address different needs but they also complicates the choice of the 'perfect' method for answering a specific question. Here I will briefly describe the principles of super-resolution optical microscopy techniques and then focus on comparing their characteristics in various aspects of practical applications.

  20. 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.

  1. Optical Property Analyses of Plant Cells for Adaptive Optics Microscopy

    NASA Astrophysics Data System (ADS)

    Tamada, Yosuke; Murata, Takashi; Hattori, Masayuki; Oya, Shin; Hayano, Yutaka; Kamei, Yasuhiro; Hasebe, Mitsuyasu

    2014-04-01

    In astronomy, adaptive optics (AO) can be used to cancel aberrations caused by atmospheric turbulence and to perform diffraction-limited observation of astronomical objects from the ground. AO can also be applied to microscopy, to cancel aberrations caused by cellular structures and to perform high-resolution live imaging. As a step toward the application of AO to microscopy, here we analyzed the optical properties of plant cells. We used leaves of the moss Physcomitrella patens, which have a single layer of cells and are thus suitable for optical analysis. Observation of the cells with bright field and phase contrast microscopy, and image degradation analysis using fluorescent beads demonstrated that chloroplasts provide the main source of optical degradations. Unexpectedly, the cell wall, which was thought to be a major obstacle, has only a minor effect. Such information provides the basis for the application of AO to microscopy for the observation of plant cells.

  2. 3D shape measurement of optical free-form surface based on fringe projection

    NASA Astrophysics Data System (ADS)

    Li, Shaohui; Liu, Shugui; Zhang, Hongwei

    2011-05-01

    Present a novel method of 3D shape measurement of optical free-from surface based on fringe projection. A virtual reference surface is proposed which can be used to improve the detection efficiency and realize the automation of measuring process. Sinusoidal fringe patterns are projected to the high reflected surface of the measured object. The deflection fringe patterns that modulated by the object surface are captured by the CCD camera. The slope information can be obtained by analyzing the relationship between the phase deflectometry and the slope of the object surface. The wave-front reconstruction method is used to reconstruct the surface. With the application of fringe projection technology the accuracy of optical free-form surfaces measurement could reach the level of tens of micrometer or even micrometer.

  3. Planned development of a 3D computer based on free-space optical interconnects

    NASA Astrophysics Data System (ADS)

    Neff, John A.; Guarino, David R.

    1994-05-01

    Free-space optical interconnection has the potential to provide upwards of a million data channels between planes of electronic circuits. This may result in the planar board and backplane structures of today giving away to 3-D stacks of wafers or multi-chip modules interconnected via channels running perpendicular to the processor planes, thereby eliminating much of the packaging overhead. Three-dimensional packaging is very appealing for tightly coupled fine-grained parallel computing where the need for massive numbers of interconnections is severely taxing the capabilities of the planar structures. This paper describes a coordinated effort by four research organizations to demonstrate an operational fine-grained parallel computer that achieves global connectivity through the use of free space optical interconnects.

  4. Respiratory motion correction in 3-D PET data with advanced optical flow algorithms.

    PubMed

    Dawood, Mohammad; Buther, Florian; Jiang, Xiaoyi; Schafers, Klaus P

    2008-08-01

    The problem of motion is well known in positron emission tomography (PET) studies. The PET images are formed over an elongated period of time. As the patients cannot hold breath during the PET acquisition, spatial blurring and motion artifacts are the natural result. These may lead to wrong quantification of the radioactive uptake. We present a solution to this problem by respiratory-gating the PET data and correcting the PET images for motion with optical flow algorithms. The algorithm is based on the combined local and global optical flow algorithm with modifications to allow for discontinuity preservation across organ boundaries and for application to 3-D volume sets. The superiority of the algorithm over previous work is demonstrated on software phantom and real patient data.

  5. 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

  6. 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.

  7. 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.

  8. 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.

  9. Optical lens-shift design for increasing spatial resolution of 3D ToF cameras

    NASA Astrophysics Data System (ADS)

    Lietz, Henrik; Hassan, M. Muneeb; Eberhardt, Jörg

    2017-02-01

    Sensor resolution of 3D time-of-flight (ToF) outdoor-capable cameras is strongly limited because of its large pixel dimensions. Computational imaging permits enhancement of the optical system's resolving power without changing physical sensor properties. Super-resolution (SR) algorithms superimpose several sub-pixel-shifted low-resolution (LR) images to overcome the system's limited spatial sampling rate. In this paper, we propose a novel opto-mechanical system to implement sub-pixel shifts by moving an optical lens. This method is more flexible in terms of implementing SR techniques than current sensor-shift approaches. In addition, we describe a SR observation model that has been optimized for the use of LR 3D ToF cameras. A state-of-the-art iteratively reweighted minimization algorithm executes the SR process. It is proven that our method achieves nearly the same resolution increase as if the pixel area would be halved physically. Resolution enhancement is measured objectively for amplitude images of a static object scene.

  10. Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI).

    PubMed

    Dertinger, T; Colyer, R; Iyer, G; Weiss, S; Enderlein, J

    2009-12-29

    Super-resolution optical microscopy is a rapidly evolving area of fluorescence microscopy with a tremendous potential for impacting many fields of science. Several super-resolution methods have been developed over the last decade, all capable of overcoming the fundamental diffraction limit of light. We present here an approach for obtaining subdiffraction limit optical resolution in all three dimensions. This method relies on higher-order statistical analysis of temporal fluctuations (caused by fluorescence blinking/intermittency) recorded in a sequence of images (movie). We demonstrate a 5-fold improvement in spatial resolution by using a conventional wide-field microscope. This resolution enhancement is achieved in iterative discrete steps, which in turn allows the evaluation of images at different resolution levels. Even at the lowest level of resolution enhancement, our method features significant background reduction and thus contrast enhancement and is demonstrated on quantum dot-labeled microtubules of fibroblast cells.

  11. 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

  12. Optical microscopy beyond the diffraction limit

    PubMed Central

    Smolyaninov, Igor I.

    2008-01-01

    Over the past century the resolution of far-field optical microscopes, which rely on propagating optical modes, was widely believed to be limited because of diffraction to a value on the order of a half-wavelength λ∕2 of the light used. Although immersion microscopes had slightly improved resolution on the order of λ∕2n, the increased resolution was limited by the small range of refractive indices, n, of available transparent materials. We are experiencing quick demolition of the diffraction limit in optical microscopy. Over the past few years numerous nonlinear optical microscopy techniques based on photoswitching and saturation of fluorescence demonstrated far-field resolution of 20 to 30 nm. The latest exciting example of these techniques has been demonstrated by Huang et al. [Science 319, 810–813 (2008)]. Moreover, recent progress in metamaterials indicates that artificial optical media can be created, which do not exhibit the diffraction limit. Resolution of linear “immersion” microscopes based on such metamaterials appears limited only by losses, which can be compensated by gain media. Thus, optical microscopy is quickly moving towards the 10 nm resolution scale, which should bring about numerous revolutionary advances in biomedical imaging. PMID:19404465

  13. High resolution 3D imaging of living cells with sub-optical wavelength phonons

    NASA Astrophysics Data System (ADS)

    Pérez-Cota, Fernando; Smith, Richard J.; Moradi, Emilia; Marques, Leonel; Webb, Kevin F.; Clark, Matt

    2016-12-01

    Label-free imaging of living cells below the optical diffraction limit poses great challenges for optical microscopy. Biologically relevant structural information remains below the Rayleigh limit and beyond the reach of conventional microscopes. Super-resolution techniques are typically based on the non-linear and stochastic response of fluorescent labels which can be toxic and interfere with cell function. In this paper we present, for the first time, imaging of live cells using sub-optical wavelength phonons. The axial imaging resolution of our system is determined by the acoustic wavelength (λa = λprobe/2n) and not on the NA of the optics allowing sub-optical wavelength acoustic sectioning of samples using the time of flight. The transverse resolution is currently limited to the optical spot size. The contrast mechanism is significantly determined by the mechanical properties of the cells and requires no additional contrast agent, stain or label to image the cell structure. The ability to breach the optical diffraction limit to image living cells acoustically promises to bring a new suite of imaging technologies to bear in answering exigent questions in cell biology and biomedicine.

  14. High resolution 3D imaging of living cells with sub-optical wavelength phonons

    PubMed Central

    Pérez-Cota, Fernando; Smith, Richard J.; Moradi, Emilia; Marques, Leonel; Webb, Kevin F.; Clark, Matt

    2016-01-01

    Label-free imaging of living cells below the optical diffraction limit poses great challenges for optical microscopy. Biologically relevant structural information remains below the Rayleigh limit and beyond the reach of conventional microscopes. Super-resolution techniques are typically based on the non-linear and stochastic response of fluorescent labels which can be toxic and interfere with cell function. In this paper we present, for the first time, imaging of live cells using sub-optical wavelength phonons. The axial imaging resolution of our system is determined by the acoustic wavelength (λa = λprobe/2n) and not on the NA of the optics allowing sub-optical wavelength acoustic sectioning of samples using the time of flight. The transverse resolution is currently limited to the optical spot size. The contrast mechanism is significantly determined by the mechanical properties of the cells and requires no additional contrast agent, stain or label to image the cell structure. The ability to breach the optical diffraction limit to image living cells acoustically promises to bring a new suite of imaging technologies to bear in answering exigent questions in cell biology and biomedicine. PMID:27996028

  15. 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

  16. 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.

  17. Multiphoton microscopy of engineered dermal substitutes: assessment of 3D collagen matrix remodeling induced by fibroblasts contraction

    NASA Astrophysics Data System (ADS)

    Pena, A.-M.; Olive, C.; Michelet, J.-F.; Galey, J.-B.; Fagot, D.; Leroy, F.; Martin, J.-L.; Colonna, A.; Schanne-Klein, M.-C.

    2010-02-01

    One of the main functions of dermal fibroblasts is the generation of mechanical forces within their surrounding extracellular matrix. Investigating molecules that could modulate fibroblast contraction and act as potent anti aging ingredients requires the development of three-dimensional in situ imaging methodologies for dermal substitute analysis. Here we use multiphoton microscopy in order to investigate the fibroblast-induced collagen matrix reorganization in engineered dermal tissue and to evaluate the effect of Y27632, a RhoA kinase inhibitor on dermal substitutes contraction. We observe that collagen fibrils rearrange around fibroblast with increasing density in control samples, whereas collagen fibrils show no remodeling in the samples containing the RhoA kinase inhibitor. Moreover, when the culture medium containing the inhibitor was replaced with a control medium, the dermal substitutes presented the same 3D reorganization as the control samples, which indicates that the inhibitory effects are reversible. In conclusion, our study demonstrates the relevance of multiphoton microscopy to visualize three-dimensional remodeling of the matrix induced by fibroblast contraction.

  18. A 3D High Frequency Array Based 16 Channel Photoacoustic Microscopy System for In Vivo Micro-vascular Imaging

    PubMed Central

    Zemp, Roger; Yen, Jesse; Wang, L.V.; Shung, K. Kirk

    2009-01-01

    This paper discusses the design of a novel photoacoustic microscopy imaging system with promise for studying the structure of tissue microvasculature for applications in visualizing angiogenesis. A new sixteen channel analog and digital high frequency array based photoacoustic microscopy system (PAM) was developed using an Nd:YLF pumped tunable dye laser, a 30MHz piezo composite linear array transducer and a custom multi-channel receiver electronics system. Using offline delay and sum beamforming and beamsteering, phantom images were obtained from a 6µm carbon fiber in water at a depth of 8mm. The measured -6dB lateral and axial spatial resolution of the system was 100±5µm and 45±5µm, respectively. The dynamic focusing capability of the system was demonstrated by imaging a composite carbon fiber matrix through a 12.5mm imaging depth. Next, 2-D in vivo images were formed of vessels around 100µm in diameter in the human hand. 3-D in vivo images were also formed of micro-vessels 3mm below the surface of the skin in two Sprague Dawley rats. PMID:19131292

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. 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

  4. A novel lithography process for 3D (three-dimensional) interconnect using an optical direct-writing exposure system

    NASA Astrophysics Data System (ADS)

    Azuma, T.; Sekiguchi, M.; Matsuo, M.; Kawasaki, A.; Hagiwara, K.; Matsui, H.; Kawamura, N.; Kishimoto, K.; Nakamura, A.; Washio, Y.

    2010-03-01

    A novel lithography process for 3D (Three-dimensional) interconnect was developed using an optical direct-writing exposure tool. A reflective IR (Infra-red) alignment system allows a direct detection of alignment marks both on front-side and back-side of wafer, and consequently allows feasible micro-fabrication for 3D interconnect using the reversed wafer. A combination of the optical direct-writing exposure tool of Dainippon Screen MFG. Co., Ltd. with the reflective IR alignment system and a high aspect chemically amplified resist of Tokyo Ohka Kogyo Co., Ltd. provides the lithography process exclusively for 12-inch wafer level 3D interconnect.

  5. Axial-Stereo 3-D Optical Metrology for Inner Profile of Pipes Using a Scanning Laser Endoscope.

    PubMed

    Gong, Yuanzheng; Johnston, Richard S; Melville, C David; Seibel, Eric J

    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.

  6. 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

  7. Structure-Function Studies of Blood and Air Capillaries in Chicken Lung Using 3D Electron Microscopy

    PubMed Central

    West, John B.; Fu, Zhenxing; Deerinck, Thomas J.; Mackey, Mason R.; Obayashi, James T.; Ellisman, Mark H.

    2010-01-01

    Avian pulmonary capillaries differ from those of mammals in three important ways. The blood-gas barrier is much thinner, it is more uniform in thickness, and the capillaries are far more rigid when their transmural pressure is altered. The thinness of the barrier is surprising because it predisposes the capillaries to stress failure. A possible mechanism for these differences is that avian pulmonary capillaries, unlike mammalian, are supported from the outside by air capillaries, but the details of the support are poorly understood. To clarify this we studied the blood and air capillaries in chicken lung using transmission electron microscopy (EM) and two relatively new techniques that allow 3D visualization: electron tomography and serial block-face scanning EM. These studies show that the pulmonary capillaries are flanked by epithelial bridges composed of two extremely thin epithelial cells with large surface areas. The junctions of the bridges with the capillary walls show thickening of the epithelial cells and an accumulation of extracellular matrix. Collapse of the pulmonary capillaries when the pressure outside them is increased is apparently prevented by the guy wire-like action of the epithelial bridges. The enlarged junctions between the bridges and the walls could provide a mechanism that limits the hoop stress in the capillary walls when the pressure inside them is increased. The support of the pulmonary capillaries may also be explained by an interdependence mechanism whereby the capillaries are linked to a rigid assemblage of air capillaries. These EM studies show the supporting structures in greater detail than has previously been possible, particularly in 3D, and they allow a more complete analysis of the mechanical forces affecting avian pulmonary capillaries. PMID:20038456

  8. 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.

  9. Automated Sensor for 3-D Reconstruction of Optical Emission from RF Plasmas

    NASA Astrophysics Data System (ADS)

    Collard, Corey; Shannon, S.; Brake, M. L.; Holloway, James Paul

    1999-10-01

    Three dimensional images are obtained by using an automated scanning sensor which collects optical emission from a RF (13.56 MHz) discharge in a capacitively coupled GEC cell. The sensor scans a plane parallel to the electrode surface and transmits the plasma spectral emission through a fiber optic cable to a monochromator. The fiber optic is attached to a motorized rotational stage attached to a manual vertical translational stage. Wedges of light (argon at 750.4 nm) are collected as the fiber scans across the plasma. The data is digitized and stored so that it can be input into an algorithm, which uses a Tikhonov regularization method to reconstruct the emissivity as a function of radial position. By varying the height of the sensor, a 3-D plot of the plasma emission can be obtained. Three dimensional plots of plasmas run at 75, 100, 150 and 200 peak to peak voltage at pressures of 100, 250, 500 and 1000 mTorr were obtained. The non-uniformity of the light emission as a function of pressure and power will be discussed.

  10. Optical security and anti-counterfeiting using 3D screen printing

    NASA Astrophysics Data System (ADS)

    Wu, W. H.; Yang, W. K.; Cheng, S. H.; Kuo, M. K.; Lee, H. W.; Chang, C. C.; Jeng, G. R.; Liu, C. P.

    2007-04-01

    This work presents a novel method for optical decrypted key production by screen printing technology. The key is mainly used to decrypt encoded information hidden inside documents containing Moire patterns and integral photographic 3D auto-stereoscopic images as a second-line security file. The proposed method can also be applied as an anti-counterfeiting measure in artistic screening. Decryption is performed by matching the correct angle between the decoding key and the document with a text or a simple geometric pattern. This study presents the theoretical analysis and experimental results of the decoded key production by the best parameter combination of Moire pattern size and screen printing elements. Experimental results reveal that the proposed method can be applied in anti-counterfeit document design for the fast and low-cost production of decryption key.

  11. Full optical characterization of autostereoscopic 3D displays using local viewing angle and imaging measurements

    NASA Astrophysics Data System (ADS)

    Boher, Pierre; Leroux, Thierry; Bignon, Thibault; Collomb-Patton, Véronique

    2012-03-01

    Two commercial auto-stereoscopic 3D displays are characterized a using Fourier optics viewing angle system and an imaging video-luminance-meter. One display has a fixed emissive configuration and the other adapts its emission to the observer position using head tracking. For a fixed emissive condition, three viewing angle measurements are performed at three positions (center, right and left). Qualified monocular and binocular viewing spaces in front of the display are deduced as well as the best working distance. The imaging system is then positioned at this working distance and crosstalk homogeneity on the entire surface of the display is measured. We show that the crosstalk is generally not optimized on all the surface of the display. Display aspect simulation using viewing angle measurements allows understanding better the origin of those crosstalk variations. Local imperfections like scratches and marks generally increase drastically the crosstalk, demonstrating that cleanliness requirements for this type of display are quite critical.

  12. 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.

  13. Intra-retinal layer segmentation of 3D optical coherence tomography using coarse grained diffusion map.

    PubMed

    Kafieh, Raheleh; Rabbani, Hossein; Abramoff, Michael D; Sonka, Milan

    2013-12-01

    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.

  14. 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.

  15. 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

  16. Correction of Depth-Dependent Aberrations in 3D Single Molecule Localization and Super-resolution Microscopy

    PubMed Central

    McGorty, Ryan; Schnitzbauer, Joerg; Zhang, Wei; Huang, Bo

    2014-01-01

    Single molecule switching based super-resolution microscopy techniques have been extended into three dimensions through various 3D single molecule localization methods. However, the localization accuracy in z can be severely degraded by the presence of aberrations, particularly the spherical aberration introduced by the refractive-index-mismatch when imaging into an aqueous sample with an oil immersion objective. This aberration confines the imaging depth in most experiments to regions close to the coverslip. Here, we show a method to obtain accurate, depth dependent z calibrations by measuring the point spread function (PSF) at the coverslip surface, calculating the microscope pupil function through phase retrieval, and then computing the depth dependent PSF with the addition of spherical aberrations. We demonstrate experimentally that this method can maintain z localization accuracy over a large range of imaging depths. Our super-resolution images of a mammalian cell nucleus acquired between 0 and 2.5 μm past the coverslip show that this method produces accurate z localizations even in the deepest focal plane. PMID:24562125

  17. Building 3D aerial image in photoresist with reconstructed mask image acquired with optical microscope

    NASA Astrophysics Data System (ADS)

    Chou, C. S.; Tang, Y. P.; Chu, F. S.; Huang, W. C.; Liu, R. G.; Gau, T. S.

    2012-03-01

    Calibration of mask images on wafer becomes more important as features shrink. Two major types of metrology have been commonly adopted. One is to measure the mask image with scanning electron microscope (SEM) to obtain the contours on mask and then simulate the wafer image with optical simulator. The other is to use an optical imaging tool Aerial Image Measurement System (AIMSTM) to emulate the image on wafer. However, the SEM method is indirect. It just gathers planar contours on a mask with no consideration of optical characteristics such as 3D topography structures. Hence, the image on wafer is not predicted precisely. Though the AIMSTM method can be used to directly measure the intensity at the near field of a mask but the image measured this way is not quite the same as that on the wafer due to reflections and refractions in the films on wafer. Here, a new approach is proposed to emulate the image on wafer more precisely. The behavior of plane waves with different oblique angles is well known inside and between planar film stacks. In an optical microscope imaging system, plane waves can be extracted from the pupil plane with a coherent point source of illumination. Once plane waves with a specific coherent illumination are analyzed, the partially coherent component of waves could be reconstructed with a proper transfer function, which includes lens aberration, polarization, reflection and refraction in films. It is a new method that we can transfer near light field of a mask into an image on wafer without the disadvantages of indirect SEM measurement such as neglecting effects of mask topography, reflections and refractions in the wafer film stacks. Furthermore, with this precise latent image, a separated resist model also becomes more achievable.

  18. 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.

  19. Optical diffraction microscopy in a teaching laboratory

    NASA Astrophysics Data System (ADS)

    Thibault, Pierre; Rankenburg, Ivan C.

    2007-09-01

    We discuss an optics experiment that reproduces all important aspects of diffraction microscopy or coherent diffractive imaging. This technique is used to reconstruct an object's image from its diffraction pattern. The experimental setup is described in detail and only requires material readily available in a well-equipped optics teaching laboratory. The data analysis procedure is explained, in particular the reconstruction part, for which an iterative phase retrieval algorithm is used. The method is illustrated by showing the complex-valued reconstruction of an insect wing from a diffraction pattern measured with this setup.

  20. 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.

  1. Beam position controlling method for 3D optical system and its application in non-planar ring resonators.

    PubMed

    Yuan, Jie; Chen, Meixiong; Long, Xingwu; Tan, Yanyang; Kang, Zhenglong; Li, Yingying

    2012-08-13

    A novel theoretical beam position controlling method for 3D optical system has been proposed in this paper. Non-planar ring resonator, which is a typical 3D optical system, has been chosen as an example to show its application. To the best of our knowledge, the generalized ray matrices, augmented 5 × 5 ray matrices for paraxial dielectric interface transmission and paraxial optical-wedge transmission, and their detailed deducing process have been proposed in this paper for the first time. By utilizing the novel coordinate system for Gaussian beam reflection and the generalized ray matrix of paraxial optical-wedge transmission, the rules and some novel results of the optical-axis perturbations of non-planar ring resonators have been obtained. Wedge angle-induced mismatching errors of non-planar ring resonators have been found out and two experimental beam position controlling methods to effectively eliminate the wedge angle-induced mismatching errors have been proposed. All those results have been confirmed by related alignment experiments and the experimental results have been described with diagrammatic representation. These findings are important to the beam control, cavity design, and cavity alignment of high precision non-planar ring laser gyroscopes. Those generalized ray matrices and their deducing methods are valuable for ray analysis of various kinds of paraxial optical-elements and resonators. This novel theoretical beam position controlling method for 3D optical system is valuable for the controlling of various kinds of 3D optical systems.

  2. 3-D imaging mass spectrometry of protein distributions in mouse Neurofibromatosis 1 (NF1)-associated optic glioma.

    PubMed

    Anderson, David M G; Van de Plas, Raf; Rose, Kristie L; Hill, Salisha; Schey, Kevin L; Solga, Anne C; Gutmann, David H; Caprioli, Richard M

    2016-10-21

    Neurofibromatosis type 1 (NF1) is a common neurogenetic disorder, in which affected individuals develop tumors of the nervous system. Children with NF1 are particularly prone to brain tumors (gliomas) involving the optic pathway that can result in impaired vision. Since tumor formation and expansion requires a cooperative tumor microenvironment, it is important to identify the cellular and acellular components associated with glioma development and growth. In this study, we used 3-D matrix assisted laser desorption ionization imaging mass spectrometry (MALDI IMS) to measure the distributions of multiple molecular species throughout optic nerve tissue in mice with and without glioma, and to explore their spatial relationships within the 3-D volume of the optic nerve and chiasm. 3-D IMS studies often involve extensive workflows due to the high volume of sections required to generate high quality 3-D images. Herein, we present a workflow for 3-D data acquisition and volume reconstruction using mouse optic nerve tissue. The resulting 3-D IMS data yield both molecular similarities and differences between glioma-bearing and wild-type (WT) tissues, including protein distributions localizing to different anatomical subregions.

  3. 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.

  4. Label-free optical detection of cells grown in 3D silicon microstructures.

    PubMed

    Merlo, Sabina; Carpignano, Francesca; Silva, Gloria; Aredia, Francesca; Scovassi, A Ivana; Mazzini, Giuliano; Surdo, Salvatore; Barillaro, Giuseppe

    2013-08-21

    We demonstrate high aspect-ratio photonic crystals that could serve as three-dimensional (3D) microincubators for cell culture and also provide label-free optical detection of the cells. The investigated microstructures, fabricated by electrochemical micromachining of standard silicon wafers, consist of periodic arrays of silicon walls separated by narrow deeply etched air-gaps (50 μm high and 5 μm wide) and feature the typical spectral properties of photonic crystals in the wavelength range 1.0-1.7 μm: their spectral reflectivity is characterized by wavelength regions where reflectivity is high (photonic bandgaps), separated by narrow wavelength regions where reflectivity is very low. In this work, we show that the presence of cells, grown inside the gaps, strongly affects light propagation across the photonic crystal and, therefore, its spectral reflectivity. Exploiting a label-free optical detection method, based on a fiberoptic setup, we are able to probe the extension of cells adherent to the vertical silicon walls with a non-invasive direct testing. In particular, the intensity ratio at two wavelengths is the experimental parameter that can be well correlated to the cell spreading on the silicon wall inside the gaps.

  5. 3D imaging of tomato seeds using frequency domain optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Fan, Chuanmao; Yao, Gang

    2012-05-01

    A fast imaging system that can reveal internal sample structures is important for research and quality controls of seeds. Optical coherence tomography (OCT) is a non-invasive optical imaging technique that can acquire high speed, high resolution depth-resolved images in scattering samples. It has found numerous applications in studying various biological tissues and other materials in vivo. A few studies have reported the use of OCT in studying seed morphology. However, 3D imaging of internal seed structure has not been reported before. In this study, we used a frequency domain OCT system to image tomato seeds. The system has a central wavelength of 844nm with a 46.8 nm FWHM bandwidth. The requirement for depth scan was eliminated by using a Fourier domain implementation. The B-scan imaging speed was limited by the spectroscopic imaging CCD at 52 kHz. The calibrated system has a 6.7μm depth resolution and a 15.4μm lateral resolution. Our results show that major seed structures can be clearly visualized in OCT images.

  6. 3D-optical measurement system using a new vignetting aperture procedure

    NASA Astrophysics Data System (ADS)

    Hofbauer, Engelbert; Rascher, Rolf; Wühr, Konrad; Friedke, Felix; Stubenrauch, Thomas; Pastötter, Benjamin; Schleich, Sebastian; Zöcke, Christine

    2014-05-01

    A newly developed measuring procedure uses vignetting to evaluate angles and angle changes, independently from the measurement distance. Further on, the same procedure enables the transmission of a digital readout and therefore a better automation of the electronic signal evaluation, for use as an alignment telescope. The fully extended readout by a simple 3-D reflector will provide the user with a measurement result with six degrees of freedom. The vignetting field stop procedure will be described. Firstly, considering artificial vignetting, the theoretical basics from geometric-optical view are represented. Secondly, the natural vignetting with photometric effects will be considered. The distribution of intensity in the image plane light spot, the so-called V-SPOT, is analytically deduced as a function of differently measured variables. Intensity shifts within the V-Spot are examined independently from different effects by numeric simulation. On these basics, the theoretical research regarding accuracy, linearity as well as results in 2 dimensional surface reconstruction on precision optical mirrors and also three dimensional measurements in mechanical engineering are examined. Effects and deviations will be discussed. The project WiPoVi is sponsored by "Ingenieur Nachwuchs - Qualifizierung von Ingenieurnachwuchs an Fachhochschulen" by Bavarian State Ministry of Education, Science and the Arts.

  7. 3D optical simulation formalism OPTOS for textured silicon solar cells.

    PubMed

    Tucher, Nico; Eisenlohr, Johannes; Kiefel, Peter; Höhn, Oliver; Hauser, Hubert; Peters, Marius; Müller, Claas; Goldschmidt, Jan Christoph; Bläsi, Benedikt

    2015-11-30

    In this paper we introduce the three-dimensional formulation of the OPTOS formalism, a matrix-based method that allows for the efficient simulation of non-coherent light propagation and absorption in thick textured sheets. As application examples, we calculate the absorptance of solar cells featuring textures on front and rear side with different feature sizes operating in different optical regimes. A discretization of polar and azimuth angle enables a three-dimensional description of systems with arbitrary surface textures. We present redistribution matrices for 3D surface textures, including pyramidal textures, binary crossed gratings and a Lambertian scatterer. The results of the OPTOS simulations for silicon sheets with different combinations of these surfaces are in accordance with both optical measurements and results based on established simulation methods like ray tracing. Using OPTOS, we show that the integration of a diffractive grating at the rear side of a silicon solar cell featuring a pyramidal front side results in absorption close to the Yablonovitch Limit enhancing the photocurrent density by 0.6 mA/cm2 for a 200 µm thick cell.

  8. 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.

  9. 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

  10. 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.

  11. Nonlinear Optical Macroscopic Assessment of 3-D Corneal Collagen Organization and Axial Biomechanics

    PubMed Central

    Winkler, Moritz; Chai, Dongyul; Kriling, Shelsea; Nien, Chyong Jy; Brown, Donald J.; Jester, Bryan; Juhasz, Tibor

    2011-01-01

    Purpose. To characterize and quantify the collagen fiber (lamellar) organization of human corneas in three dimensions by using nonlinear optical high-resolution macroscopy (NLO-HRMac) and to correlate these findings with mechanical data obtained by indentation testing of corneal flaps. Methods. Twelve corneas from 10 donors were studied. Vibratome sections, 200 μm thick, from five donor eyes were cut along the vertical meridian from limbus to limbus (arc length, 12 mm). Backscattered second harmonic–generated (SHG) NLO signals from these sections were collected as a series of overlapping 3-D images, which were concatenated to form a single 3-D mosaic (pixel resolution: 0.44 μm lateral, 2 μm axial). Collagen fiber intertwining was quantified by determining branching point density as a function of stromal depth. Mechanical testing was performed on corneal flaps from seven additional eyes. Corneas were cut into three layers (anterior, middle, and posterior) using a femtosecond surgical laser system and underwent indentation testing to determine the elastic modulus for each layer. Results. The 3-D reconstructions revealed complex collagen fiber branching patterns in the anterior cornea, with fibers extending from the anterior limiting lamina (ALL, Bowman's layer), intertwining with deeper fibers and reinserting back to the ALL, forming bow spring–like structures. Measured branching-point density was four times higher in the anterior third of the cornea than in the posterior third and decreased logarithmically with increasing distance from the ALL. Indentation testing showed an eightfold increase in elastic modulus in the anterior stroma. Conclusions. The axial gradient in lamellar intertwining appears to be associated with an axial gradient in the effective elastic modulus of the cornea, suggesting that collagen fiber intertwining and formation of bow spring–like structures provide structural support similar to cross-beams in bridges and large-scale structures

  12. Optical Measurement of Micromechanics and Structure in a 3D Fibrin Extracellular Matrix

    NASA Astrophysics Data System (ADS)

    Kotlarchyk, Maxwell Aaron

    2011-07-01

    In recent years, a significant number of studies have focused on linking substrate mechanics to cell function using standard methodologies to characterize the bulk properties of the hydrogel substrates. However, current understanding of the correlations between the microstructural mechanical properties of hydrogels and cell function in 3D is poor, in part because of a lack of appropriate techniques. Methods for tuning extracellular matrix (ECM) mechanics in 3D cell culture that rely on increasing the concentration of either protein or cross-linking molecules fail to control important parameters such as pore size, ligand density, and molecular diffusivity. Alternatively, ECM stiffness can be modulated independently from protein concentration by mechanically loading the ECM. We have developed an optical tweezers-based microrheology system to investigate the fundamental role of ECM mechanical properties in determining cellular behavior. Further, this thesis outlines the development of a novel device for generating stiffness gradients in naturally derived ECMs, where stiffness is tuned by inducing strain, while local structure and mechanical properties are directly determined by laser tweezers-based passive and active microrheology respectively. Hydrogel substrates polymerized within 35 mm diameter Petri dishes are strained non-uniformly by the precise rotation of an embedded cylindrical post, and exhibit a position-dependent stiffness with little to no modulation of local mesh geometry. Here we present microrheological studies in the context of fibrin hydrogels. Microrheology and confocal imaging were used to directly measure local changes in micromechanics and structure respectively in unstrained hydrogels of increasing fibrinogen concentration, as well as in our strain gradient device, in which the concentration of fibrinogen is held constant. Orbital particle tracking, and raster image correlation analysis are used to quantify changes in fibrin mechanics on the

  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.

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

    PubMed

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

    2011-08-01

    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 mm(2). 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.

  15. X-Ray and Optical Videography for 3D Measurement of Capillary and Melt Pool Geometry in Laser Welding

    NASA Astrophysics Data System (ADS)

    Boley, M.; Abt, F.; Weber, R.; Graf, T.

    This paper describes a method to reconstruct the 3D shape of the melt pool and the capillary of a laser keyhole welding process. Three different diagnostic methods, including X-Ray and optical videography as well as metallographic cross sections are combined to gain the three dimensional data of the solidus-liquidus-surface. A detailed description of the experimental setup and a discussion of different methods to combine the 2D data sets of the three different diagnostic methods to a 3D-model will be given. The result will be a static 3D description of the welding process.

  16. 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

  17. 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

  18. 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.

  19. Threshold response using modulated continuous wave illumination for multilayer 3D optical data storage

    NASA Astrophysics Data System (ADS)

    Saini, A.; Christenson, C. W.; Khattab, T. A.; Wang, R.; Twieg, R. J.; Singer, K. D.

    2017-01-01

    In order to achieve a high capacity 3D optical data storage medium, a nonlinear or threshold writing process is necessary to localize data in the axial dimension. To this end, commercial multilayer discs use thermal ablation of metal films or phase change materials to realize such a threshold process. This paper addresses a threshold writing mechanism relevant to recently reported fluorescence-based data storage in dye-doped co-extruded multilayer films. To gain understanding of the essential physics, single layer spun coat films were used so that the data is easily accessible by analytical techniques. Data were written by attenuating the fluorescence using nanosecond-range exposure times from a 488 nm continuous wave laser overlapping with the single photon absorption spectrum. The threshold writing process was studied over a range of exposure times and intensities, and with different fluorescent dyes. It was found that all of the dyes have a common temperature threshold where fluorescence begins to attenuate, and the physical nature of the thermal process was investigated.

  20. Globular and Optically Transparent Photonic Crystals Based on 3D-opal Matrix and REE

    NASA Astrophysics Data System (ADS)

    Ivicheva, S. N.; Kargin, Yu. F.; Gorelik, V. S.

    By repeatedly filling the octahedral and tetrahedral pores of 3D-silica opal matrices with silica sol doped with rare-earth elements with subsequent heat treatment globular photonic crystals filled with mesoporous glass and optically transparent photonic crystals (quantytes) containing 10-30 ppm REE were produced, depending on the annealing temperature. Voids of fcc lattice formed by amorphous spherical globules of SiO2 in globular photonic crystals are filled (up to 70%) by mesoporous glass doped with rare earth elements. Pores in the transparent photonic crystals disappear during sintering of globules of silica and mesoporous glass, but the periodic arrangement of REE-enriched silica areas (quantum dots) is retained. The reflection and luminescence spectra of photonic crystals filled with sols doped with europium Eu3+ and terbium Tb3+ were experimentally studied. A significant increase in the photoluminescence intensity of Eu3+ ions at the approach of the spectral position of the transition 5D0 → 7F2 to the edge of the bandgaps of the photonic crystal was determined. The authors come to the conclusion that a lowering of the threshold for lasing transitions in ions of rare elements is possible.

  1. 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.

  2. 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.

  3. Optical analysis of nanoparticles via enhanced backscattering facilitated by 3-D photonic nanojets.

    PubMed

    Li, Xu; Chen, Zhigang; Taflove, Allen; Backman, Vadim

    2005-01-24

    We report the phenomenon of ultra-enhanced backscattering of visible light by nanoparticles facilitated by the 3-D photonic nanojet - a sub-diffraction light beam appearing at the shadow side of a plane-waveilluminated dielectric microsphere. Our rigorous numerical simulations show that backscattering intensity of nanoparticles can be enhanced up to eight orders of magnitude when locating in the nanojet. As a result, the enhanced backscattering from a nanoparticle with diameter on the order of 10 nm is well above the background signal generated by the dielectric microsphere itself. We also report that nanojet-enhanced backscattering is extremely sensitive to the size of the nanoparticle, permitting in principle resolving sub-nanometer size differences using visible light. Finally, we show how the position of a nanoparticle could be determined with subdiffractional accuracy by recording the angular distribution of the backscattered light. These properties of photonic nanojets promise to make this phenomenon a useful tool for optically detecting, differentiating, and sorting nanoparticles.

  4. Optical analysis of nanoparticles via enhanced backscattering facilitated by 3-D photonic nanojets

    NASA Astrophysics Data System (ADS)

    Li, Xu; Chen, Zhigang; Taflove, Allen; Backman, Vadim

    2005-01-01

    We report the phenomenon of ultra-enhanced backscattering of visible light by nanoparticles facilitated by the 3-D photonic nanojet a sub-diffraction light beam appearing at the shadow side of a plane-waveilluminated dielectric microsphere. Our rigorous numerical simulations show that backscattering intensity of nanoparticles can be enhanced up to eight orders of magnitude when locating in the nanojet. As a result, the enhanced backscattering from a nanoparticle with diameter on the order of 10 nm is well above the background signal generated by the dielectric microsphere itself. We also report that nanojet-enhanced backscattering is extremely sensitive to the size of the nanoparticle, permitting in principle resolving sub-nanometer size differences using visible light. Finally, we show how the position of a nanoparticle could be determined with subdiffractional accuracy by recording the angular distribution of the backscattered light. These properties of photonic nanojets promise to make this phenomenon a useful tool for optically detecting, differentiating, and sorting nanoparticles.

  5. Comparative validation of single-shot optical techniques for laparoscopic 3-D surface reconstruction.

    PubMed

    Maier-Hein, L; Groch, A; Bartoli, A; Bodenstedt, S; Boissonnat, G; Chang, P-L; Clancy, N T; Elson, D S; Haase, S; Heim, E; Hornegger, J; Jannin, P; Kenngott, H; Kilgus, T; Müller-Stich, B; Oladokun, D; Röhl, S; Dos Santos, T R; Schlemmer, H-P; Seitel, A; Speidel, S; Wagner, M; Stoyanov, D

    2014-10-01

    Intra-operative imaging techniques for obtaining the shape and morphology of soft-tissue surfaces in vivo are a key enabling technology for advanced surgical systems. Different optical techniques for 3-D surface reconstruction in laparoscopy have been proposed, however, so far no quantitative and comparative validation has been performed. Furthermore, robustness of the methods to clinically important factors like smoke or bleeding has not yet been assessed. To address these issues, we have formed a joint international initiative with the aim of validating different state-of-the-art passive and active reconstruction methods in a comparative manner. In this comprehensive in vitro study, we investigated reconstruction accuracy using different organs with various shape and texture and also tested reconstruction robustness with respect to a number of factors like the pose of the endoscope as well as the amount of blood or smoke present in the scene. The study suggests complementary advantages of the different techniques with respect to accuracy, robustness, point density, hardware complexity and computation time. While reconstruction accuracy under ideal conditions was generally high, robustness is a remaining issue to be addressed. Future work should include sensor fusion and in vivo validation studies in a specific clinical context. To trigger further research in surface reconstruction, stereoscopic data of the study will be made publically available at www.open-CAS.com upon publication of the paper.

  6. Combining 3D optical imaging and dual energy absorptiometry to measure three compositional components.

    PubMed

    Malkov, Serghei; Shepherd, John

    2014-02-17

    We report on the design of the technique combining 3D optical imaging and dual-energy absorptiometry body scanning to estimate local body area compositions of three compartments. Dual-energy attenuation and body shape measures are used together to solve for the three compositional tissue thicknesses: water, lipid, and protein. We designed phantoms with tissue-like properties as our reference standards for calibration purposes. The calibration was created by fitting phantom values using non-linear regression of quadratic and truncated polynomials. Dual-energy measurements were performed on tissue-mimicking phantoms using a bone densitometer unit. The phantoms were made of materials shown to have similar x-ray attenuation properties of the biological compositional compartments. The components for the solid phantom were tested and their high energy/low energy attenuation ratios are in good correspondent to water, lipid, and protein for the densitometer x-ray region. The three-dimensional body shape was reconstructed from the depth maps generated by Microsoft Kinect for Windows. We used open-source Point Cloud Library and freeware software to produce dense point clouds. Accuracy and precision of compositional and thickness measures were calculated. The error contributions due to two modalities were estimated. The preliminary phantom composition and shape measurements are found to demonstrate the feasibility of the method proposed.

  7. 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

  8. 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.

  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. Nonlinear Optical Microscopy of Single Nanostructures

    NASA Astrophysics Data System (ADS)

    Huang, Libai; Cheng, Ji-Xin

    2013-07-01

    We review recent advances in nonlinear optical (NLO) microscopy studies of single nanostructures. NLO signals are intrinsically sensitive to the electronic, vibrational, and structural properties of such nanostructures. Ultrafast excitation allows for mapping of energy relaxation pathways at the single-particle level. The strong nonlinear response of nanostructures makes them highly attractive for applications as novel NLO imaging agents in biological and biomedical research. NLO modalities based on harmonic generation, multiphoton photoluminescence, four-wave mixing, and pump-probe processes are discussed in detail.

  13. Optical Third-Harmonic Microscopy of Graphene

    NASA Astrophysics Data System (ADS)

    Dadap, Jerry I.; Hong, Sung-Young; Petrone, Nicholas W.; Yeh, Po-Chun; Hone, James C.; Osgood, Richard M., Jr.

    2013-03-01

    We report strong third-harmonic (TH) generation in monolayer graphene mounted on an amorphous silica substrate using a photon energy that is three-photon resonant with the exciton-shifted van Hove singularity at the M-point of graphene. Our polarization-dependent and azimuthal rotation measurements confirm the expected isotropic symmetry properties for the TH nonlinear optical process in graphene. Since this monolayer graphene TH signal exceeds that of bulk glass by more than two orders of magnitude, the signal contrast permits background-free scanning of graphene and provides structural information that is difficult to obtain via linear optical microscopy. We also discuss the dependence of TH signals on the number of graphene layers and compare the graphene signal strength with that from crystalline Au(111) sample. We acknowledge support from AFOSR MURI Program #FA9550-09-1-0705.

  14. 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

  15. 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.

  16. 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.

  17. DMD-based LED-illumination super-resolution and optical sectioning microscopy.

    PubMed

    Dan, Dan; Lei, Ming; Yao, Baoli; Wang, Wen; Winterhalder, Martin; Zumbusch, Andreas; Qi, Yujiao; Xia, Liang; Yan, Shaohui; Yang, Yanlong; Gao, Peng; Ye, Tong; Zhao, Wei

    2013-01-01

    Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×10(7) pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

  18. Optical oscillator strengths for valence-shell and Br-3d inner-shell excitations of HCl and HBr.

    PubMed

    Li, Wen-Bin; Zhu, Lin-Fan; Yuan, Zhen-Sheng; Liu, Xiao-Jing; Xu, Ke-Zun

    2006-10-21

    Absolute optical oscillator strength density spectra for valence-shell excitations of HCl and HBr, as well as for Br-3d inner-shell excitations of HBr, have been determined by high-resolution electron-energy-loss-spectroscopy method in the dipole limit. Absolute optical oscillator strengths for the discrete transitions of HCl and HBr are reported and compared with the previous results determined by the photoabsorption method.

  19. 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.

  20. Assessment of Iterative Closest Point Registration Accuracy for Different Phantom Surfaces Captured by an Optical 3D Sensor in Radiotherapy

    PubMed Central

    Walke, Mathias; Gademann, Günther

    2017-01-01

    An optical 3D sensor provides an additional tool for verification of correct patient settlement on a Tomotherapy treatment machine. The patient's position in the actual treatment is compared with the intended position defined in treatment planning. A commercially available optical 3D sensor measures parts of the body surface and estimates the deviation from the desired position without markers. The registration precision of the in-built algorithm and of selected ICP (iterative closest point) algorithms is investigated on surface data of specially designed phantoms captured by the optical 3D sensor for predefined shifts of the treatment table. A rigid body transform is compared with the actual displacement to check registration reliability for predefined limits. The curvature type of investigated phantom bodies has a strong influence on registration result which is more critical for surfaces of low curvature. We investigated the registration accuracy of the optical 3D sensor for the chosen phantoms and compared the results with selected unconstrained ICP algorithms. Safe registration within the clinical limits is only possible for uniquely shaped surface regions, but error metrics based on surface normals improve translational registration. Large registration errors clearly hint at setup deviations, whereas small values do not guarantee correct positioning. PMID:28163773

  1. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

    PubMed Central

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-01-01

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format. PMID:27886235

  2. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

    NASA Astrophysics Data System (ADS)

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-01

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  3. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.

    PubMed

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-25

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  4. 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.

  5. Analysis of 3D-printed metal for rapid-prototyped reflective terahertz optics

    NASA Astrophysics Data System (ADS)

    Headland, Daniel; Withayachumnankul, Withawat; Webb, Michael; Ebendorff-Heidepriem, Heike; Luiten, Andre; Abbott, Derek

    2016-07-01

    We explore the potential of 3D metal printing to realize complex conductive terahertz devices. Factors impacting performance such as printing resolution, surface roughness, oxidation, and material loss are investigated via analytical, numerical, and experimental approaches. The high degree of control offered by a 3D-printed topology is exploited to realize a zone plate operating at 530 GHz. Reflection efficiency at this frequency is found to be over 90%. The high-performance of this preliminary device suggest that 3D metal printing can play a strong role in guided-wave and general beam control devices in the terahertz range.

  6. 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.

  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. 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.

  9. Doppler encoded excitation pattern tomographic optical microscopy.

    PubMed

    Feldkhun, Daniel; Wagner, Kelvin H

    2010-12-01

    Most far-field optical imaging systems rely on lenses and spatially resolved detection to probe distinct locations on the object. We describe and demonstrate a high-speed wide-field approach to imaging that instead measures the complex spatial Fourier transform of the object by detecting its spatially integrated response to dynamic acousto-optically synthesized structured illumination. Tomographic filtered backprojection is applied to reconstruct the object in two or three dimensions. This technique decouples depth of field and working distance from resolution, in contrast to conventional imaging, and can be used to image biological and synthetic structures in fluoresced or scattered light employing coherent or broadband illumination. We discuss the electronically programmable transfer function of the optical system and its implications for imaging dynamic processes. We also explore wide-field fluorescence imaging in scattering media by coherence gating. Finally, we present two-dimensional high-resolution tomographic image reconstructions in both scattered and fluoresced light demonstrating a thousandfold improvement in the depth of field compared to conventional lens-based microscopy.

  10. Optical low-cost and portable arrangement for full field 3D displacement measurement using a single camera

    NASA Astrophysics Data System (ADS)

    López-Alba, E.; Felipe-Sesé, L.; Schmeer, S.; Díaz, F. A.

    2016-11-01

    In the current paper, an optical low-cost system for 3D displacement measurement based on a single camera and 3D digital image correlation is presented. The conventional 3D-DIC set-up based on a two-synchronized-cameras system is compared with a proposed pseudo-stereo portable system that employs a mirror system integrated in a device for a straightforward application achieving a novel handle and flexible device for its use in many scenarios. The proposed optical system splits the image by the camera into two stereo images of the object. In order to validate this new approach and quantify its uncertainty compared to traditional 3D-DIC systems, solid rigid in and out-of-plane displacements experiments have been performed and analyzed. The differences between both systems have been studied employing an image decomposition technique which performs a full image comparison. Therefore, results of all field of view are compared with those using a stereoscopy system and 3D-DIC, discussing the accurate results obtained with the proposed device not having influence any distortion or aberration produced by the mirrors. Finally, the adaptability of the proposed system and its accuracy has been tested performing quasi-static and dynamic experiments using a silicon specimen under high deformation. Results have been compared and validated with those obtained from a conventional stereoscopy system showing an excellent level of agreement.

  11. 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).

  12. 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

  13. Integrated optical coherence tomography and optical coherence microscopy imaging of human pathology

    NASA Astrophysics Data System (ADS)

    Lee, Hsiang-Chieh; Zhou, Chao; Wang, Yihong; Aquirre, Aaron D.; Tsai, Tsung-Han; Cohen, David W.; Connolly, James L.; Fujimoto, James G.

    2010-02-01

    Excisional biopsy is the current gold standard for disease diagnosis; however, it requires a relatively long processing time and it may also suffer from unacceptable false negative rates due to sampling errors. Optical coherence tomography (OCT) is a promising imaging technique that provide real-time, high resolution and three-dimensional (3D) images of tissue morphology. Optical coherence microscopy (OCM) is an extension of OCT, combining both the coherence gating and the confocal gating techniques. OCM imaging achieves cellular resolution with deeper imaging depth compared to confocal microscopy. An integrated OCT/OCM imaging system can provide co-registered multiscale imaging of tissue morphology. 3D-OCT provides architectural information with a large field of view and can be used to find regions of interest; while OCM provides high magnification to enable cellular imaging. The integrated OCT/OCM system has an axial resolution of <4um and transverse resolutions of 14um and <2um for OCT and OCM, respectively. In this study, a wide range of human pathologic specimens, including colon (58), thyroid (43), breast (34), and kidney (19), were imaged with OCT and OCM within 2 to 6 hours after excision. The images were compared with H & E histology to identify characteristic features useful for disease diagnosis. The feasibility of visualizing human pathology using integrated OCT/OCM was demonstrated in the pathology laboratory settings.

  14. 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

  15. A 3D Optical Surface Profilometer Using a Dual-Frequency Liquid Crystal-Based Dynamic Fringe Pattern Generator

    PubMed Central

    Joo, Kyung-Il; Kim, Mugeon; Park, Min-Kyu; Park, Heewon; Kim, Byeonggon; Hahn, JoonKu; Kim, Hak-Rin

    2016-01-01

    We propose a liquid crystal (LC)-based 3D optical surface profilometer that can utilize multiple fringe patterns to extract an enhanced 3D surface depth profile. To avoid the optical phase ambiguity and enhance the 3D depth extraction, 16 interference patterns were generated by the LC-based dynamic fringe pattern generator (DFPG) using four-step phase shifting and four-step spatial frequency varying schemes. The DFPG had one common slit with an electrically controllable birefringence (ECB) LC mode and four switching slits with a twisted nematic LC mode. The spatial frequency of the projected fringe pattern could be controlled by selecting one of the switching slits. In addition, moving fringe patterns were obtainable by applying voltages to the ECB LC layer, which varied the phase difference between the common and the selected switching slits. Notably, the DFPG switching time required to project 16 fringe patterns was minimized by utilizing the dual-frequency modulation of the driving waveform to switch the LC layers. We calculated the phase modulation of the DFPG and reconstructed the depth profile of 3D objects using a discrete Fourier transform method and geometric optical parameters. PMID:27801812

  16. A new optimization approach for the calibration of an ultrasound probe using a 3D optical localizer.

    PubMed

    Dardenne, G; Cano, J D Gil; Hamitouche, C; Stindel, E; Roux, C

    2007-01-01

    This paper describes a fast procedure for the calibration of an ultrasound (US) probe using a 3D optical localizer. This calibration step allows us to obtain the 3D position of any point located on the 2D ultrasonic (US) image. To carry out correctly this procedure, a phantom of known geometric properties is probed and these geometries are found in the US images. A segmentation step is applied in order to obtain automatically the needed information in the US images and then, an optimization approach is performed to find the optimal calibration parameters. A new optimization method to estimate the calibration parameters for an ultrasound (US) probe is developed.

  17. FreeCAD visualization of realistic 3D physical optics beams within a CAD system-model

    NASA Astrophysics Data System (ADS)

    Gayer, D.; O'Sullivan, C.; Scully, S.; Burke, D.; Brossard, J.; Chapron, C.

    2016-07-01

    The facility to realise the shape and extent of optical beams within a telescope or beamcombiner can aid greatly in the design and layout of optical elements within the system. It can also greatly facilitate communication between the optical design team and other teams working on the mechanical design of an instrument. Beyond the realm where raytracing is applicable however, it becomes much more difficult to realise accurate 3D beams which incorporate diffraction effects. It then is another issue to incorporate this into a CAD model of the system. A novel method is proposed which has been used to aid with the design of an optical beam combiner for the QUBIC (Q and U Bolometric Interferometer for Cosmology) 1 experiment operating at 150 GHz and 220 GHz. The method combines calculation work in GRASP 2, a commercial physical optics modelling tool from TICRA, geometrical work in Mathematica, and post processing in MATLAB. Finally, the Python console of the open source package FreeCAD3 is exploited to realise the 3D beams in a complete CAD system-model of the QUBIC optical beam combiner. This paper details and explains the work carried out to reach the goal and presents some graphics of the outcome. 3D representations of beams from some back-to-back input horns of the QUBIC instrument are shown within the CAD model. Beams of the -3dB and -13dB contour envelope are shown as well as envelopes enclosing 80% and 95% of the power of the beam. The ability to see these beams in situ with all the other elements of the combiner such as mirrors, cold stop, beam splitter and cryostat widows etc. greatly simplified the design for these elements and facilitated communication of element dimension and location between different subgroups within the QUBIC group.

  18. Near Field Scanning Optical Microscopy (NSOM)

    PubMed Central

    Betzig, E.; Lewis, A.; Harootunian, A.; Isaacson, M.; Kratschmer, E.

    1986-01-01

    A new method for high-resolution imaging, near-field scanning optical microscopy (NSOM), has been developed. The concepts governing this method are discussed, and the technical challenges encountered in constructing a working NSOM instrument are described. Two distinct methods are presented for the fabrication of well-characterized, highly reproducible, subwavelength apertures. A sample one-dimensional scan is provided and compared to the scanning electron micrograph of a test pattern. From this comparison, a resolution of > 1,500 Å (i.e., ≃λ/3.6) is determined, which represents a significant step towards our eventual goal of 500 Å resolution. Fluorescence has been observed through apertures smaller than 600 Å and signal-to-noise calculations show that fluorescent imaging should be feasible. The application of such imaging is then discussed in reference to specific biological problems. The NSOM method employs nonionizing visible radiation and can be used in air or aqueous environments for nondestructive visualization of functioning biological systems with a resolution comparable to that of scanning electron microscopy. ImagesFIGURE 4FIGURE 7FIGURE 9FIGURE 10 PMID:19431633

  19. Analyzing Structure and Function of Vascularization in Engineered Bone Tissue by Video-Rate Intravital Microscopy and 3D Image Processing

    PubMed Central

    Pang, Yonggang; Tsigkou, Olga; Spencer, Joel A.; Lin, Charles P.; Neville, Craig

    2015-01-01

    Vascularization is a key challenge in tissue engineering. Three-dimensional structure and microcirculation are two fundamental parameters for evaluating vascularization. Microscopic techniques with cellular level resolution, fast continuous observation, and robust 3D postimage processing are essential for evaluation, but have not been applied previously because of technical difficulties. In this study, we report novel video-rate confocal microscopy and 3D postimage processing techniques to accomplish this goal. In an immune-deficient mouse model, vascularized bone tissue was successfully engineered using human bone marrow mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs) in a poly (d,l-lactide-co-glycolide) (PLGA) scaffold. Video-rate (30 FPS) intravital confocal microscopy was applied in vitro and in vivo to visualize the vascular structure in the engineered bone and the microcirculation of the blood cells. Postimage processing was applied to perform 3D image reconstruction, by analyzing microvascular networks and calculating blood cell viscosity. The 3D volume reconstructed images show that the hMSCs served as pericytes stabilizing the microvascular network formed by HUVECs. Using orthogonal imaging reconstruction and transparency adjustment, both the vessel structure and blood cells within the vessel lumen were visualized. Network length, network intersections, and intersection densities were successfully computed using our custom-developed software. Viscosity analysis of the blood cells provided functional evaluation of the microcirculation. These results show that by 8 weeks, the blood vessels in peripheral areas function quite similarly to the host vessels. However, the viscosity drops about fourfold where it is only 0.8 mm away from the host. In summary, we developed novel techniques combining intravital microscopy and 3D image processing to analyze the vascularization in engineered bone. These techniques have broad

  20. Flying triangulation--an optical 3D sensor for the motion-robust acquisition of complex objects.

    PubMed

    Ettl, Svenja; Arold, Oliver; Yang, Zheng; Häusler, Gerd

    2012-01-10

    Three-dimensional (3D) shape acquisition is difficult if an all-around measurement of an object is desired or if a relative motion between object and sensor is unavoidable. An optical sensor principle is presented-we call it "flying triangulation"-that enables a motion-robust acquisition of 3D surface topography. It combines a simple handheld sensor with sophisticated registration algorithms. An easy acquisition of complex objects is possible-just by freely hand-guiding the sensor around the object. Real-time feedback of the sequential measurement results enables a comfortable handling for the user. No tracking is necessary. In contrast to most other eligible sensors, the presented sensor generates 3D data from each single camera image.

  1. 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.

  2. Fast optical sectioning obtained by structured illumination microscopy using a digital mirror device.

    PubMed

    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 6.9×10(7)  pixel/s of our system, an order of magnitude faster than previously reported.

  3. 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.

  4. Monitoring adipose-derived stem cells within 3D carrier by combined dielectric spectroscopy and spectral domain optical coherence topography

    NASA Astrophysics Data System (ADS)

    Bagnaninchi, P. O.

    2010-02-01

    Monitoring non-invasively the cellular events in three dimensional carriers is a major challenge for tissue engineering and regenerative medicine that prevents time-lapsed studies over large population of sample. The potential of optical coherence tomography has been demonstrated to assess tissue formation within porous matrices. In this study we explore the combination of dielectric spectroscopy (DS) and spectral domain optical coherence tomography (SDOCT) to quality assess ADSCs loaded in three dimensional carriers. A SDOCT (930nm, FWHM 90nm) was combined to an open ended coaxial probe connected to material analyser, and broadband measurements between 20MHz and 1GHz were synchronized with Labview. Both ADSCs maintained in undifferentiated state within 3D carrier and induced towards osteoblasts were monitored with this multimodality technique and their DS spectra were acquired at high cell concentration simultaneously to 3D imaging. This multimodality technique will be instrumental to assess non-invasively cell loaded carriers for cell therapy.

  5. Conventional and nonlinear optical microscopy of liquid crystal colloids

    NASA Astrophysics Data System (ADS)

    Lee, Taewoo; Smalyukh, Ivan I.

    The fast-growing field of liquid crystal colloids requires increasingly sophisticated optical microscopy tools for experimental studies. Recent technological advances have resulted in a vast body of new imaging modalities, such as nonlinear optical microscopy techniques, that were developed to achieve high resolution while probing director structures and material composition at length scales ranging from hundreds of nanometers to oscopic. These techniques are ideally suited for experimental exploration of liquid crystal colloids. The goal of this chapter is to introduce a variety of optical microscopy techniques available to researchers in the field, starting from basic principles and finishing with a discussion of the most advanced microscopy systems. We describe traditional imaging tools, such as bright field and polarizing optical microscopy, along with state-of-the-art orientationsensitive three-dimensional imaging techniques, such as various nonlinear optical microscopies. Applications of these different imaging approaches are illustrated by providing specific examples of imaging of liquid crystal colloids and other soft matter systems.

  6. Cardiac muscle organization revealed in 3-D by imaging whole-mount mouse hearts using two-photon fluorescence and confocal microscopy.

    PubMed

    Sivaguru, Mayandi; Fried, Glenn; Sivaguru, Barghav S; Sivaguru, Vignesh A; Lu, Xiaochen; Choi, Kyung Hwa; Saif, M Taher A; Lin, Brian; Sadayappan, Sakthivel

    2015-11-01

    The ability to image the entire adult mouse heart at high resolution in 3-D would provide enormous advantages in the study of heart disease. However, a technique for imaging nuclear/cellular detail as well as the overall structure of the entire heart in 3-D with minimal effort is lacking. To solve this problem, we modified the benzyl alcohol:benzyl benzoate (BABB) clearing technique by labeling mouse hearts with periodic acid Schiff (PAS) stain. We then imaged the hearts with a combination of two-photon fluorescence microscopy and automated tile-scan imaging/stitching. Utilizing the differential spectral properties of PAS, we could identify muscle and nuclear compartments in the heart. We were also able to visualize the differences between a 3-month-old normal mouse heart and a mouse heart that had undergone heart failure due to the expression of cardiac myosin binding protein-C (cMyBP-C) gene mutation (t/t). Using 2-D and 3-D morphometric analysis, we found that the t/t heart had anomalous ventricular shape, volume, and wall thickness, as well as a disrupted sarcomere pattern. We further validated our approach using decellularized hearts that had been cultured with 3T3 fibroblasts, which were tracked using a nuclear label. We were able to detect the 3T3 cells inside the decellularized intact heart tissue, achieving nuclear/cellular resolution in 3-D. The combination of labeling, clearing, and two-photon microscopy together with tiling eliminates laborious and time-consuming physical sectioning, alignment, and 3-D reconstruction.

  7. 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.

  8. 3-D confocal laser scanning microscopy used in morphometric analysis of rat Purkinje cell dendritic spines after chronic ethanol consumption.

    PubMed

    Wenisch, S; Fortmann, B; Steinmetz, T; Kriete, A; Leiser, R; Bitsch, I

    1998-12-01

    A confocal laser scanning microscope (with a 543 nm laser) was used for imaging rat Purkinje cell dendritic spines at high 3-D resolution. In a nutritionally controlled study of the rat, 5 months of ethanol consumption was demonstrated to alter the spines of Purkinje cell dendrites in rat cerebellum. Intact spines showed significant elongation after ethanol exposure, whereas this neuromorphological alteration could not be detected in controls. Spine elongation could be regarded as compensative growth of spines in search of new synaptic contacts due to alcohol induced cell loss.

  9. A 3D imaging and visualization workflow, using confocal microscopy and advanced image processing for brachyuran crab larvae.

    PubMed

    Kamanli, S A; Kihara, T C; Ball, A D; Morritt, D; Clark, P F

    2017-03-07

    Confocal laser scanning microscopy is an excellent tool for nondestructive imaging of arthropods and can provide detailed information on morphology including fine surface detail. A methodology is presented here for the visualization by confocal microscopy of arthropods, using brachyuran crab zoeal stages as examples and postprocessing techniques derived from micro-CT protocols to improve the final images. This protocol is divided into description of the preprocessing steps (cleaning, staining, digesting and mounting), confocal laser scanning microscopy and data visualization using open-source, freeware programs ImageJ and Drishti. The advantages of using ImageJ to standardize stack data and Drishti for surface rendering are discussed. The methodology has been comprehensively tested using data acquired from all four brands of confocal microscope (Leica, Nikon, Olympus and Zeiss).

  10. Towards a Noninvasive Intracranial Tumor Irradiation Using 3D Optical Imaging and Multimodal Data Registration

    PubMed Central

    Posada, R.; Daul, Ch.; Wolf, D.; Aletti, P.

    2007-01-01

    Conformal radiotherapy (CRT) results in high-precision tumor volume irradiation. In fractioned radiotherapy (FRT), lesions are irradiated in several sessions so that healthy neighbouring tissues are better preserved than when treatment is carried out in one fraction. In the case of intracranial tumors, classical methods of patient positioning in the irradiation machine coordinate system are invasive and only allow for CRT in one irradiation session. This contribution presents a noninvasive positioning method representing a first step towards the combination of CRT and FRT. The 3D data used for the positioning is point clouds spread over the patient's head (CT-data usually acquired during treatment) and points distributed over the patient's face which are acquired with a structured light sensor fixed in the therapy room. The geometrical transformation linking the coordinate systems of the diagnosis device (CT-modality) and the 3D sensor of the therapy room (visible light modality) is obtained by registering the surfaces represented by the two 3D point sets. The geometrical relationship between the coordinate systems of the 3D sensor and the irradiation machine is given by a calibration of the sensor position in the therapy room. The global transformation, computed with the two previous transformations, is sufficient to predict the tumor position in the irradiation machine coordinate system with only the corresponding position in the CT-coordinate system. Results obtained for a phantom show that the mean positioning error of tumors on the treatment machine isocentre is 0.4 mm. Tests performed with human data proved that the registration algorithm is accurate (0.1 mm mean distance between homologous points) and robust even for facial expression changes. PMID:18364992

  11. Writing of 3D optical integrated circuits with ultrashort laser pulses in the presence of strong spherical aberration

    NASA Astrophysics Data System (ADS)

    Bukharin, M. A.; Skryabin, N. N.; Khudyakov, D. V.; Vartapetov, S. K.

    2016-09-01

    A novel technique was proposed for 3D femtosecond writing of waveguides and optical integrated circuits in the presence of strong spherical aberration, caused by inscription at significantly different depth under the surface of optical glasses and crystals. Strong negative effect of spherical aberration and related asymmetry of created structures was reduced due to transition to the cumulative thermal regime of femtosecond interaction with the material. The differences in the influence of spherical aberration effect in a broad depth range (larger than 200 µm) was compensated by dynamic adjustment of laser pulse energy during the process of waveguides recording. The presented approach has been experimentally implemented in fused silica. Obtained results can be used in production of a broad class of femtosecond written three-dimensional integrated optical systems, inscripted at non-optimal (for focusing lens) optical depth or in significantly extended range of depths.

  12. Fabrication 3D buried channel optical waveguide modulators on field-driven ion exchange process

    NASA Astrophysics Data System (ADS)

    Zhou, Zigang; Chen, Wenqiang; Zhu, Li; Li, Jing; Luo, Xiaoying

    2010-10-01

    A high electric field technique was developed to fabricate buried optical waveguide modulator on K9 optical glass. The 80V voltage was applied on the glass to accelerate the field-driven ion exchange process by expeditiously replacing host sodium ions in the glass with silver ions. As a result, the optical loss for optical waveguide modulator was measured using the edge coupling technique with a 0.6328μm He-Ne laser. Loss of 0.20 dB/cm was obtained for channel waveguides of 25μm in depth, relatively low for waveguides of such depth at red wavelength.

  13. Label-free subcellular 3D live imaging of preimplantation mouse embryos with full-field optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Zheng, Jing-gao; Lu, Danyu; Chen, Tianyuan; Wang, Chengming; Tian, Ning; Zhao, Fengying; Huo, Tiancheng; Zhang, Ning; Chen, Dieyan; Ma, Wanyun; Sun, Jia-lin; Xue, Ping

    2012-07-01

    Early patterning and polarity is of fundamental interest in preimplantation embryonic development. Label-free subcellular 3D live imaging is very helpful to its related studies. We have developed a novel system of full-field optical coherence tomography (FF-OCT) for noninvasive 3D subcellular live imaging of preimplantation mouse embryos with no need of dye labeling. 3D digitized embryos can be obtained by image processing. Label-free 3D live imaging is demonstrated for the mouse embryos at various typical preimplantation stages with a spatial resolution of 0.7 μm and imaging rate of 24 fps. Factors that relate to early patterning and polarity, such as pronuclei in zygote, shapes of zona pellucida, location of second polar body, cleavage planes, and the blastocyst axis, can be quantitatively measured. The angle between the two second cleavage planes is accurately measured to be 87 deg. It is shown that FF-OCT provides a potential breakthrough for early patterning, polarity formation, and many other preimplantation-related studies in mammalian developmental biology.

  14. Unsaturated lipid bodies as a hallmark of inflammation studied by Raman 2D and 3D microscopy

    NASA Astrophysics Data System (ADS)

    Czamara, K.; Majzner, K.; Selmi, A.; Baranska, M.; Ozaki, Y.; Kaczor, A.

    2017-01-01

    Endothelial HMEC-1 cells incubated with pro-inflammatory cytokine TNF-α for 6 and 24 hours were studied as a model of inflammation using Raman imaging. Striking changes in distribution, composition and concentration of cellular lipids were observed after exposure to TNF-α compared to the control. In particular, 3D Raman imaging revealed a significant increase in the amount of lipid entities formed under inflammation. Lipid bodies were randomly distributed in the cytoplasm and two types of droplets were assembled: more saturated one, in spectral characteristics resembling phosphatidylcholine and saturated cholesteryl esters, observed also in the control, and highly unsaturated one, containing also cholesterols, being a hallmark of inflamed cells. The statistical analysis showed that the number of lipid bodies was significantly dependent on the exposure time to TNF-α. Overall, observed formation of unsaturated lipid droplets can be directly correlated with the increase in production of prostacyclins - endogenous inflammation mediators.