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Sample records for 3d microscopy images

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

    PubMed

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

    2015-03-01

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

  2. 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.

  3. 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

  4. 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

  5. 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.

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

    PubMed

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

    2016-06-01

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

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

    PubMed

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

    2014-11-01

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

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2015-07-01

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

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

    PubMed

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

    2008-08-01

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

  13. 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.

  14. 3D imaging of the cleared intact murine colon with light sheet microscopy

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

  15. 3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy.

    PubMed

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

    2014-08-01

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

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

    PubMed

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

    2015-07-17

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

  17. 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.

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

    NASA Astrophysics Data System (ADS)

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

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

  19. 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

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

    NASA Astrophysics Data System (ADS)

    Duocastella, Marti; Vicidomini, Giuseppe; Diaspro, Alberto

    2015-03-01

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

  1. 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

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

    PubMed

    Franzini-Armstrong, Clara

    2015-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-05-01

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

  4. 3D elemental sensitive imaging using transmission X-ray microscopy.

    PubMed

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

    2012-09-01

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

  5. 3D elemental sensitive imaging using transmission X-ray microscopy.

    PubMed

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

    2012-09-01

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

  6. 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

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

    PubMed

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

    2014-01-01

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

  8. 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.

  9. 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.

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

    PubMed

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

    2016-01-01

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

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

    PubMed

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

    2014-09-01

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

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2009-01-01

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

  15. 3D Imaging.

    ERIC Educational Resources Information Center

    Hastings, S. K.

    2002-01-01

    Discusses 3 D imaging as it relates to digital representations in virtual library collections. Highlights include X-ray computed tomography (X-ray CT); the National Science Foundation (NSF) Digital Library Initiatives; output peripherals; image retrieval systems, including metadata; and applications of 3 D imaging for libraries and museums. (LRW)

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

    PubMed

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

    2015-05-01

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

  17. 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.

  18. 3DSEM: A 3D microscopy dataset.

    PubMed

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

    2016-03-01

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

  19. 3DSEM: A 3D microscopy dataset

    PubMed Central

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

    2015-01-01

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

  20. 3DSEM: A 3D microscopy dataset.

    PubMed

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

    2016-03-01

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

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

    NASA Astrophysics Data System (ADS)

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

    1990-05-01

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

  2. TeraStitcher - A tool for fast automatic 3D-stitching of teravoxel-sized microscopy images

    PubMed Central

    2012-01-01

    Background Further advances in modern microscopy are leading to teravoxel-sized tiled 3D images at high resolution, thus increasing the dimension of the stitching problem of at least two orders of magnitude. The existing software solutions do not seem adequate to address the additional requirements arising from these datasets, such as the minimization of memory usage and the need to process just a small portion of data. Results We propose a free and fully automated 3D Stitching tool designed to match the special requirements coming out of teravoxel-sized tiled microscopy images that is able to stitch them in a reasonable time even on workstations with limited resources. The tool was tested on teravoxel-sized whole mouse brain images with micrometer resolution and it was also compared with the state-of-the-art stitching tools on megavoxel-sized publicy available datasets. This comparison confirmed that the solutions we adopted are suited for stitching very large images and also perform well on datasets with different characteristics. Indeed, some of the algorithms embedded in other stitching tools could be easily integrated in our framework if they turned out to be more effective on other classes of images. To this purpose, we designed a software architecture which separates the strategies that use efficiently memory resources from the algorithms which may depend on the characteristics of the acquired images. Conclusions TeraStitcher is a free tool that enables the stitching of Teravoxel-sized tiled microscopy images even on workstations with relatively limited resources of memory (<8 GB) and processing power. It exploits the knowledge of approximate tile positions and uses ad-hoc strategies and algorithms designed for such very large datasets. The produced images can be saved into a multiresolution representation to be efficiently retrieved and processed. We provide TeraStitcher both as standalone application and as plugin of the free software Vaa3D. PMID:23181553

  3. 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

  4. 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.

  5. 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

  6. 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

  7. 3D multiplexed immunoplasmonics microscopy.

    PubMed

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

    2016-07-21

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

  8. Efficient Semi-Automatic 3D Segmentation for Neuron Tracing in Electron Microscopy Images

    PubMed Central

    Jones, Cory; Liu, Ting; Cohan, Nathaniel Wood; Ellisman, Mark; Tasdizen, Tolga

    2015-01-01

    0.1. Background In the area of connectomics, there is a significant gap between the time required for data acquisition and dense reconstruction of the neural processes contained in the same dataset. Automatic methods are able to eliminate this timing gap, but the state-of-the-art accuracy so far is insufficient for use without user corrections. If completed naively, this process of correction can be tedious and time consuming. 0.2. New Method We present a new semi-automatic method that can be used to perform 3D segmentation of neurites in EM image stacks. It utilizes an automatic method that creates a hierarchical structure for recommended merges of superpixels. The user is then guided through each predicted region to quickly identify errors and establish correct links. 0.3. Results We tested our method on three datasets with both novice and expert users. Accuracy and timing were compared with published automatic, semi-automatic, and manual results. 0.4. Comparison with Existing Methods Post-automatic correction methods have also been used in [1] and [2]. These methods do not provide navigation or suggestions in the manner we present. Other semi-automatic methods require user input prior to the automatic segmentation such as [3] and [4] and are inherently different than our method. 0.5. Conclusion Using this method on the three datasets, novice users achieved accuracy exceeding state-of-the-art automatic results, and expert users achieved accuracy on par with full manual labeling but with a 70% time improvement when compared with other examples in publication. PMID:25769273

  9. Deconvolution in 3-D optical microscopy.

    PubMed

    Shaw, P

    1994-09-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2015-07-15

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

  12. 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

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

    NASA Astrophysics Data System (ADS)

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

    2010-05-01

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

  14. Imaging the three orientation variants of the DO22 phase by 3D atom probe microscopy.

    PubMed

    Marteau, L; Pareige, C; Blavette, D

    2001-12-01

    Three-phase NiAlV alloys were investigated using a three-dimensional atom probe. Ageing at 800 degrees C gives rise to the precipitation of two ordered phases within the supersaturated FCC solid solution, namely Ni3Al (L1(2) structure) and Ni3V (DO22 structure). The DO22 phase has three orientation variants which need to be identified in 3DAP images. It is shown that an appropriate choice of analysis site enables us to image the chemical order within both L1(2) and DO22 ordered phases and to distinguish the three orientation variants of the DO22 phase in reconstructed images. The lateral resolution of 3DAP in these experimental conditions was estimated through simple considerations to be less than 0.3 nm.

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

    NASA Astrophysics Data System (ADS)

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

    2008-03-01

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

  16. A workflow to process 3D+time microscopy images of developing organisms and reconstruct their cell lineage.

    PubMed

    Faure, Emmanuel; Savy, Thierry; Rizzi, Barbara; Melani, Camilo; Stašová, Olga; Fabrèges, Dimitri; Špir, Róbert; Hammons, Mark; Čúnderlík, Róbert; Recher, Gaëlle; Lombardot, Benoît; Duloquin, Louise; Colin, Ingrid; Kollár, Jozef; Desnoulez, Sophie; Affaticati, Pierre; Maury, Benoît; Boyreau, Adeline; Nief, Jean-Yves; Calvat, Pascal; Vernier, Philippe; Frain, Monique; Lutfalla, Georges; Kergosien, Yannick; Suret, Pierre; Remešíková, Mariana; Doursat, René; Sarti, Alessandro; Mikula, Karol; Peyriéras, Nadine; Bourgine, Paul

    2016-01-01

    The quantitative and systematic analysis of embryonic cell dynamics from in vivo 3D+time image data sets is a major challenge at the forefront of developmental biology. Despite recent breakthroughs in the microscopy imaging of living systems, producing an accurate cell lineage tree for any developing organism remains a difficult task. We present here the BioEmergences workflow integrating all reconstruction steps from image acquisition and processing to the interactive visualization of reconstructed data. Original mathematical methods and algorithms underlie image filtering, nucleus centre detection, nucleus and membrane segmentation, and cell tracking. They are demonstrated on zebrafish, ascidian and sea urchin embryos with stained nuclei and membranes. Subsequent validation and annotations are carried out using Mov-IT, a custom-made graphical interface. Compared with eight other software tools, our workflow achieved the best lineage score. Delivered in standalone or web service mode, BioEmergences and Mov-IT offer a unique set of tools for in silico experimental embryology. PMID:26912388

  17. 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

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

    PubMed Central

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

    2011-01-01

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

  19. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2015-10-01

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

  1. 3D imaging and quantitative analysis of small solubilized membrane proteins and their complexes by transmission electron microscopy.

    PubMed

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

    2013-02-01

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

  2. 3D microscopy - new powerful tools in geomaterials characterization

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

  3. 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

  4. 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.

  5. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice.

    PubMed

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated.

  6. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

    NASA Astrophysics Data System (ADS)

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, Yongkeun

    2016-09-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated.

  7. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

    PubMed Central

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated. PMID:27605489

  8. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice.

    PubMed

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated. PMID:27605489

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

    PubMed

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

    2015-10-01

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

  10. FluoRender: An Application of 2D Image Space Methods for 3D and 4D Confocal Microscopy Data Visualization in Neurobiology Research

    PubMed Central

    Wan, Yong; Otsuna, Hideo; Chien, Chi-Bin; Hansen, Charles

    2013-01-01

    2D image space methods are processing methods applied after the volumetric data are projected and rendered into the 2D image space, such as 2D filtering, tone mapping and compositing. In the application domain of volume visualization, most 2D image space methods can be carried out more efficiently than their 3D counterparts. Most importantly, 2D image space methods can be used to enhance volume visualization quality when applied together with volume rendering methods. In this paper, we present and discuss the applications of a series of 2D image space methods as enhancements to confocal microscopy visualizations, including 2D tone mapping, 2D compositing, and 2D color mapping. These methods are easily integrated with our existing confocal visualization tool, FluoRender, and the outcome is a full-featured visualization system that meets neurobiologists’ demands for qualitative analysis of confocal microscopy data. PMID:23584131

  11. FluoRender: An Application of 2D Image Space Methods for 3D and 4D Confocal Microscopy Data Visualization in Neurobiology Research.

    PubMed

    Wan, Yong; Otsuna, Hideo; Chien, Chi-Bin; Hansen, Charles

    2012-01-01

    2D image space methods are processing methods applied after the volumetric data are projected and rendered into the 2D image space, such as 2D filtering, tone mapping and compositing. In the application domain of volume visualization, most 2D image space methods can be carried out more efficiently than their 3D counterparts. Most importantly, 2D image space methods can be used to enhance volume visualization quality when applied together with volume rendering methods. In this paper, we present and discuss the applications of a series of 2D image space methods as enhancements to confocal microscopy visualizations, including 2D tone mapping, 2D compositing, and 2D color mapping. These methods are easily integrated with our existing confocal visualization tool, FluoRender, and the outcome is a full-featured visualization system that meets neurobiologists' demands for qualitative analysis of confocal microscopy data.

  12. 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.

  13. High-Resolution 3D Imaging and Quantification of Gold Nanoparticles in a Whole Cell Using Scanning Transmission Ion Microscopy

    PubMed Central

    Chen, Xiao; Chen, Ce-Belle; Udalagama, Chammika N.B.; Ren, Minqin; Fong, Kah Ee; Yung, Lin Yue Lanry; Giorgia, Pastorin; Bettiol, Andrew Anthony; Watt, Frank

    2013-01-01

    Increasing interest in the use of nanoparticles (NPs) to elucidate the function of nanometer-sized assemblies of macromolecules and organelles within cells, and to develop biomedical applications such as drug delivery, labeling, diagnostic sensing, and heat treatment of cancer cells has prompted investigations into novel techniques that can image NPs within whole cells and tissue at high resolution. Using fast ions focused to nanodimensions, we show that gold NPs (AuNPs) inside whole cells can be imaged at high resolution, and the precise location of the particles and the number of particles can be quantified. High-resolution density information of the cell can be generated using scanning transmission ion microscopy, enhanced contrast for AuNPs can be achieved using forward scattering transmission ion microscopy, and depth information can be generated from elastically backscattered ions (Rutherford backscattering spectrometry). These techniques and associated instrumentation are at an early stage of technical development, but we believe there are no physical constraints that will prevent whole-cell three-dimensional imaging at <10 nm resolution. PMID:23561518

  14. Autofocus for 3D imaging

    NASA Astrophysics Data System (ADS)

    Lee-Elkin, Forest

    2008-04-01

    Three dimensional (3D) autofocus remains a significant challenge for the development of practical 3D multipass radar imaging. The current 2D radar autofocus methods are not readily extendable across sensor passes. We propose a general framework that allows a class of data adaptive solutions for 3D auto-focus across passes with minimal constraints on the scene contents. The key enabling assumption is that portions of the scene are sparse in elevation which reduces the number of free variables and results in a system that is simultaneously solved for scatterer heights and autofocus parameters. The proposed method extends 2-pass interferometric synthetic aperture radar (IFSAR) methods to an arbitrary number of passes allowing the consideration of scattering from multiple height locations. A specific case from the proposed autofocus framework is solved and demonstrates autofocus and coherent multipass 3D estimation across the 8 passes of the "Gotcha Volumetric SAR Data Set" X-Band radar data.

  15. 3D microscopy for microfabrication quality control

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

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

  16. 3D digital image processing for biofilm quantification from confocal laser scanning microscopy: Multidimensional statistical analysis of biofilm modeling

    NASA Astrophysics Data System (ADS)

    Zielinski, Jerzy S.

    The dramatic increase in number and volume of digital images produced in medical diagnostics, and the escalating demand for rapid access to these relevant medical data, along with the need for interpretation and retrieval has become of paramount importance to a modern healthcare system. Therefore, there is an ever growing need for processed, interpreted and saved images of various types. Due to the high cost and unreliability of human-dependent image analysis, it is necessary to develop an automated method for feature extraction, using sophisticated mathematical algorithms and reasoning. This work is focused on digital image signal processing of biological and biomedical data in one- two- and three-dimensional space. Methods and algorithms presented in this work were used to acquire data from genomic sequences, breast cancer, and biofilm images. One-dimensional analysis was applied to DNA sequences which were presented as a non-stationary sequence and modeled by a time-dependent autoregressive moving average (TD-ARMA) model. Two-dimensional analyses used 2D-ARMA model and applied it to detect breast cancer from x-ray mammograms or ultrasound images. Three-dimensional detection and classification techniques were applied to biofilm images acquired using confocal laser scanning microscopy. Modern medical images are geometrically arranged arrays of data. The broadening scope of imaging as a way to organize our observations of the biophysical world has led to a dramatic increase in our ability to apply new processing techniques and to combine multiple channels of data into sophisticated and complex mathematical models of physiological function and dysfunction. With explosion of the amount of data produced in a field of biomedicine, it is crucial to be able to construct accurate mathematical models of the data at hand. Two main purposes of signal modeling are: data size conservation and parameter extraction. Specifically, in biomedical imaging we have four key problems

  17. High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens

    NASA Astrophysics Data System (ADS)

    Philipp, Katrin; Smolarski, André; Fischer, Andreas; Koukourakis, Nektarios; Stürmer, Moritz; Wallrabe, Ulricke; Czarske, Jürgen

    2016-04-01

    We present a HiLo microscope with an electrically tunable lens for high-contrast three-dimensional image acquisition. HiLo microscopy combines wide field and speckled illumination images to create optically sectioned images. Additionally, the depth-of-field is not fixed, but can be adjusted between wide field and confocal-like axial resolution. We incorporate an electrically tunable lens in the HiLo microscope for axial scanning, to obtain three-dimensional data without the need of moving neither the sample nor the objective. The used adaptive lens consists of a transparent polydimethylsiloxane (PDMS) membrane into which an annular piezo bending actuator is embedded. A transparent fluid is filled between the membrane and the glass substrate. When actuated, the piezo generates a pressure in the lens which deflects the membrane and thus changes the refractive power. This technique enables a large tuning range of the refractive power between 1/f = (-24 . . . 25) 1/m. As the NA of the adaptive lens is only about 0.05, a fixed high-NA lens is included in the setup to provide high resolution. In this contribution, the scan properties and capabilities of the tunable lens in the HiLo microscope are analyzed. Eventually, exemplary measurements are presented and discussed.

  18. 3D imaging of cells and tissues by focused ion beam/scanning electron microscopy (FIB/SEM).

    PubMed

    Drobne, Damjana

    2013-01-01

    Integration of a scanning electron microscope (SEM) and focused ion beam (FIB) technology into a single FIB/SEM system permits use of the FIB as a nano-scalpel to reveal site-specific subsurface microstructures which can be examined in great detail by SEM. The FIB/SEM technology is widely used in the semiconductor industry and material sciences, and recently its use in the life sciences has been initiated. Samples for FIB/SEM investigation can be either embedded in a plastic matrix, the traditional means of preparation of transmission electron microscopy (TEM) specimens, or simply dried as in samples prepared for SEM imaging. Currently, FIB/SEM is used in the life sciences for (a) preparation by the lift-out technique of lamella for TEM analysis, (b) tomography of samples embedded in a matrix, and (c) in situ site-specific FIB milling and SEM imaging using a wide range of magnifications. Site-specific milling and imaging has attracted wide interest as a technique in structural research of single eukaryotic and prokaryotic cells, small animals, and different animal tissue, but it still remains to be explored more thoroughly. In the past, preparation of samples for site-specific milling and imaging by FIB/SEM has typically adopted the embedding techniques used for TEM samples, and which have been very well described in the literature. Sample preparation protocols for the use of dried samples in FIB/SEM have been less well investigated. The aim of this chapter is to encourage application of FIB/SEM on dried biological samples. A detailed description of conventional dried sample preparation and FIB/SEM investigation of dried biological samples is presented. The important steps are described and illustrated, and direct comparison between embedded and dried samples of same tissues is provided. The ability to discover links between gross morphology of the tissue or organ, surface characteristics of any selected region, and intracellular structural details on the nanometer

  19. 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.

  20. Shaping Field for 3D Laser Scanning Microscopy

    PubMed Central

    Colon, Jorge; Lim, Hyungsik

    2015-01-01

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

  1. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2011-08-01

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

  3. A Bright Fluorescent Probe for H2S Enables Analyte-Responsive, 3D Imaging in Live Zebrafish Using Light Sheet Fluorescence Microscopy

    PubMed Central

    2015-01-01

    Hydrogen sulfide (H2S) is a critical gaseous signaling molecule emerging at the center of a rich field of chemical and biological research. As our understanding of the complexity of physiological H2S in signaling pathways evolves, advanced chemical and technological investigative tools are required to make sense of this interconnectivity. Toward this goal, we have developed an azide-functionalized O-methylrhodol fluorophore, MeRho-Az, which exhibits a rapid >1000-fold fluorescence response when treated with H2S, is selective for H2S over other biological analytes, and has a detection limit of 86 nM. Additionally, the MeRho-Az scaffold is less susceptible to photoactivation than other commonly used azide-based systems, increasing its potential application in imaging experiments. To demonstrate the efficacy of this probe for H2S detection, we demonstrate the ability of MeRho-Az to detect differences in H2S levels in C6 cells and those treated with AOAA, a common inhibitor of enzymatic H2S synthesis. Expanding the use of MeRho-Az to complex and heterogeneous biological settings, we used MeRho-Az in combination with light sheet fluorescence microscopy (LSFM) to visualize H2S in the intestinal tract of live zebrafish. This application provides the first demonstration of analyte-responsive 3D imaging with LSFM, highlighting the utility of combining new probes and live imaging methods for investigating chemical signaling in complex multicellular systems. PMID:26061541

  4. A Bright Fluorescent Probe for H2S Enables Analyte-Responsive, 3D Imaging in Live Zebrafish Using Light Sheet Fluorescence Microscopy.

    PubMed

    Hammers, Matthew D; Taormina, Michael J; Cerda, Matthew M; Montoya, Leticia A; Seidenkranz, Daniel T; Parthasarathy, Raghuveer; Pluth, Michael D

    2015-08-19

    Hydrogen sulfide (H2S) is a critical gaseous signaling molecule emerging at the center of a rich field of chemical and biological research. As our understanding of the complexity of physiological H2S in signaling pathways evolves, advanced chemical and technological investigative tools are required to make sense of this interconnectivity. Toward this goal, we have developed an azide-functionalized O-methylrhodol fluorophore, MeRho-Az, which exhibits a rapid >1000-fold fluorescence response when treated with H2S, is selective for H2S over other biological analytes, and has a detection limit of 86 nM. Additionally, the MeRho-Az scaffold is less susceptible to photoactivation than other commonly used azide-based systems, increasing its potential application in imaging experiments. To demonstrate the efficacy of this probe for H2S detection, we demonstrate the ability of MeRho-Az to detect differences in H2S levels in C6 cells and those treated with AOAA, a common inhibitor of enzymatic H2S synthesis. Expanding the use of MeRho-Az to complex and heterogeneous biological settings, we used MeRho-Az in combination with light sheet fluorescence microscopy (LSFM) to visualize H2S in the intestinal tract of live zebrafish. This application provides the first demonstration of analyte-responsive 3D imaging with LSFM, highlighting the utility of combining new probes and live imaging methods for investigating chemical signaling in complex multicellular systems. PMID:26061541

  5. A Bright Fluorescent Probe for H2S Enables Analyte-Responsive, 3D Imaging in Live Zebrafish Using Light Sheet Fluorescence Microscopy.

    PubMed

    Hammers, Matthew D; Taormina, Michael J; Cerda, Matthew M; Montoya, Leticia A; Seidenkranz, Daniel T; Parthasarathy, Raghuveer; Pluth, Michael D

    2015-08-19

    Hydrogen sulfide (H2S) is a critical gaseous signaling molecule emerging at the center of a rich field of chemical and biological research. As our understanding of the complexity of physiological H2S in signaling pathways evolves, advanced chemical and technological investigative tools are required to make sense of this interconnectivity. Toward this goal, we have developed an azide-functionalized O-methylrhodol fluorophore, MeRho-Az, which exhibits a rapid >1000-fold fluorescence response when treated with H2S, is selective for H2S over other biological analytes, and has a detection limit of 86 nM. Additionally, the MeRho-Az scaffold is less susceptible to photoactivation than other commonly used azide-based systems, increasing its potential application in imaging experiments. To demonstrate the efficacy of this probe for H2S detection, we demonstrate the ability of MeRho-Az to detect differences in H2S levels in C6 cells and those treated with AOAA, a common inhibitor of enzymatic H2S synthesis. Expanding the use of MeRho-Az to complex and heterogeneous biological settings, we used MeRho-Az in combination with light sheet fluorescence microscopy (LSFM) to visualize H2S in the intestinal tract of live zebrafish. This application provides the first demonstration of analyte-responsive 3D imaging with LSFM, highlighting the utility of combining new probes and live imaging methods for investigating chemical signaling in complex multicellular systems.

  6. 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.

  7. 3D electron microscopy of biological nanomachines: principles and applications.

    PubMed

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

    2007-11-01

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

  8. 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

  9. 3D ultrafast ultrasound imaging in vivo.

    PubMed

    Provost, Jean; Papadacci, Clement; Arango, Juan Esteban; Imbault, Marion; Fink, Mathias; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-10-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in 3D based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32  ×  32 matrix-array probe. Its ability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3D Shear-Wave Imaging, 3D Ultrafast Doppler Imaging, and, finally, 3D Ultrafast combined Tissue and Flow Doppler Imaging. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3D Ultrafast Doppler was used to obtain 3D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, at thousands of volumes per second, the complex 3D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, as well as the 3D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3D Ultrafast Ultrasound Imaging for the 3D mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra--and inter-observer variability.

  10. 3D ultrafast ultrasound imaging in vivo

    NASA Astrophysics Data System (ADS)

    Provost, Jean; Papadacci, Clement; Esteban Arango, Juan; Imbault, Marion; Fink, Mathias; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-10-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in 3D based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32  ×  32 matrix-array probe. Its ability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3D Shear-Wave Imaging, 3D Ultrafast Doppler Imaging, and, finally, 3D Ultrafast combined Tissue and Flow Doppler Imaging. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3D Ultrafast Doppler was used to obtain 3D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, at thousands of volumes per second, the complex 3D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, as well as the 3D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3D Ultrafast Ultrasound Imaging for the 3D mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra—and inter-observer variability.

  11. 3D ultrafast ultrasound imaging in vivo.

    PubMed

    Provost, Jean; Papadacci, Clement; Arango, Juan Esteban; Imbault, Marion; Fink, Mathias; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-10-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in 3D based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32  ×  32 matrix-array probe. Its ability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3D Shear-Wave Imaging, 3D Ultrafast Doppler Imaging, and, finally, 3D Ultrafast combined Tissue and Flow Doppler Imaging. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3D Ultrafast Doppler was used to obtain 3D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, at thousands of volumes per second, the complex 3D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, as well as the 3D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3D Ultrafast Ultrasound Imaging for the 3D mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra--and inter-observer variability. PMID:25207828

  12. 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.

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

    PubMed

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

    2014-08-01

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

  14. 3D Backscatter Imaging System

    NASA Technical Reports Server (NTRS)

    Turner, D. Clark (Inventor); Whitaker, Ross (Inventor)

    2016-01-01

    Systems and methods for imaging an object using backscattered radiation are described. The imaging system comprises both a radiation source for irradiating an object that is rotationally movable about the object, and a detector for detecting backscattered radiation from the object that can be disposed on substantially the same side of the object as the source and which can be rotationally movable about the object. The detector can be separated into multiple detector segments with each segment having a single line of sight projection through the object and so detects radiation along that line of sight. Thus, each detector segment can isolate the desired component of the backscattered radiation. By moving independently of each other about the object, the source and detector can collect multiple images of the object at different angles of rotation and generate a three dimensional reconstruction of the object. Other embodiments are described.

  15. 3D Ultrafast Ultrasound Imaging In Vivo

    PubMed Central

    Provost, Jean; Papadacci, Clement; Arango, Juan Esteban; Imbault, Marion; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-01-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative real-time imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in three dimensions based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32×32 matrix-array probe. Its capability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3-D Shear-Wave Imaging, 3-D Ultrafast Doppler Imaging and finally 3D Ultrafast combined Tissue and Flow Doppler. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3-D Ultrafast Doppler was used to obtain 3-D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, for the first time, the complex 3-D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, and the 3-D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3-D Ultrafast Ultrasound Imaging for the 3-D real-time mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra- and inter-observer variability. PMID:25207828

  16. Perception of detail in 3D images

    NASA Astrophysics Data System (ADS)

    Heynderickx, Ingrid; Kaptein, Ronald

    2009-01-01

    A lot of current 3D displays suffer from the fact that their spatial resolution is lower compared to their 2D counterparts. One reason for this is that the multiple views needed to generate 3D are often spatially multiplexed. Besides this, imperfect separation of the left- and right-eye view leads to blurring or ghosting, and therefore to a decrease in perceived sharpness. However, people watching stereoscopic videos have reported that the 3D scene contained more details, compared to the 2D scene with identical spatial resolution. This is an interesting notion, that has never been tested in a systematic and quantitative way. To investigate this effect, we had people compare the amount of detail ("detailedness") in pairs of 2D and 3D images. A blur filter was applied to one of the two images, and the blur level was varied using an adaptive staircase procedure. In this way, the blur threshold for which the 2D and 3D image contained perceptually the same amount of detail could be found. Our results show that the 3D image needed to be blurred more than the 2D image. This confirms the earlier qualitative findings that 3D images contain perceptually more details than 2D images with the same spatial resolution.

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed

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

    2013-01-01

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

  20. 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

  1. Non-destructive 3D Imaging of Extraterrestrial Materials by Synchrotron X-ray Micro- tomography (XR-CMT) and Laser Confocal Scanning Microscopy (LCSM): Beyond Pretty Pictures

    NASA Astrophysics Data System (ADS)

    Ebel, D. S.; Greenberg, M.

    2009-05-01

    We report scientific results made possible only by the use these two non-destructive 3D imaging techniques. XR-CMT provides 3D image reconstructions at spatial resolutions of 1 to 17 micron/voxel edge. We use XR- CMT to locate potential melt-inclusion-bearing phenocrysts in batches of 100-200 micron lunar fire-fountain spherules; to locate and visualize the morphology of 1-2mm size, irregular, unmelted Ca-, Al-rich inclusions (CAIs) and to quantify chondrule/matrix ratios and chondrule size distributions in 6x6x20mm chunks of carbonaceous chondrites; to quantify the modal abundance of opaque phases in similar sized Martian meteorite fragments, and in individual 1-2mm diameter chondrules from chondrites. LCSM provides 3D image stacks at resolutions < 100 nm/pixel. We are the only group creating deconvolved image stacks of 100 to over 1000 micron long comet particle tracks in aerogel keystones from the Stardust mission. We present measurements of track morphology in 3D, and locate high-value particles using complementary synchrotron x- ray fluorescence (XRF) examination. We show that bench-top LCSM extracts maximum information about tracks and particles rapidly and cheaply prior to destructive disassembly. Using XR-CMT we quantify, for the first time, the volumetric abundances of metal grains in 1-2 mm diameter CR chondrite chondrules. Metal abundances vary from 1 to 37 vol.% between 8 chondrules (and more by inspection), in a meteorite with solar (chondritic) Fe/Si ratio, indicating that chondrules formed and accreted locally from bulk solar composition material. They are 'complementary' to each other in Fe/Si ratios. Void spaces in chondritic CAIs and chondrules are shown to be a primary feature, not due to plucking during sectioning. CAI morphology in 3D reveals pre-accretionary impact features, and various types of mineralogical layering, seen in 3D, reveal the formation history of these building blocks of planets and asteroids. We also quantify the x

  2. Evaluation of 3D imaging.

    PubMed

    Vannier, M W

    2000-10-01

    Interactive computer-based simulation is gaining acceptance for craniofacial surgical planning. Subjective visualization without objective measurement capability, however, severely limits the value of simulation since spatial accuracy must be maintained. This study investigated the error sources involved in one method of surgical simulation evaluation. Linear and angular measurement errors were found to be within +/- 1 mm and 1 degree. Surface match of scanned objects was slightly less accurate, with errors up to 3 voxels and 4 degrees, and Boolean subtraction methods were 93 to 99% accurate. Once validated, these testing methods were applied to objectively compare craniofacial surgical simulations to post-operative outcomes, and verified that the form of simulation used in this study yields accurate depictions of surgical outcome. However, to fully evaluate surgical simulation, future work is still required to test the new methods in sufficient numbers of patients to achieve statistically significant results. Once completely validated, simulation cannot only be used in pre-operative surgical planning, but also as a post-operative descriptor of surgical and traumatic physical changes. Validated image comparison methods can also show discrepancy of surgical outcome to surgical plan, thus allowing evaluation of surgical technique. PMID:11098409

  3. 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.

  4. 3D holoscopic video imaging system

    NASA Astrophysics Data System (ADS)

    Steurer, Johannes H.; Pesch, Matthias; Hahne, Christopher

    2012-03-01

    Since many years, integral imaging has been discussed as a technique to overcome the limitations of standard still photography imaging systems where a three-dimensional scene is irrevocably projected onto two dimensions. With the success of 3D stereoscopic movies, a huge interest in capturing three-dimensional motion picture scenes has been generated. In this paper, we present a test bench integral imaging camera system aiming to tailor the methods of light field imaging towards capturing integral 3D motion picture content. We estimate the hardware requirements needed to generate high quality 3D holoscopic images and show a prototype camera setup that allows us to study these requirements using existing technology. The necessary steps that are involved in the calibration of the system as well as the technique of generating human readable holoscopic images from the recorded data are discussed.

  5. Nonlaser-based 3D surface imaging

    SciTech Connect

    Lu, Shin-yee; Johnson, R.K.; Sherwood, R.J.

    1994-11-15

    3D surface imaging refers to methods that generate a 3D surface representation of objects of a scene under viewing. Laser-based 3D surface imaging systems are commonly used in manufacturing, robotics and biomedical research. Although laser-based systems provide satisfactory solutions for most applications, there are situations where non laser-based approaches are preferred. The issues that make alternative methods sometimes more attractive are: (1) real-time data capturing, (2) eye-safety, (3) portability, and (4) work distance. The focus of this presentation is on generating a 3D surface from multiple 2D projected images using CCD cameras, without a laser light source. Two methods are presented: stereo vision and depth-from-focus. Their applications are described.

  6. Miniaturized 3D microscope imaging system

    NASA Astrophysics Data System (ADS)

    Lan, Yung-Sung; Chang, Chir-Weei; Sung, Hsin-Yueh; Wang, Yen-Chang; Chang, Cheng-Yi

    2015-05-01

    We designed and assembled a portable 3-D miniature microscopic image system with the size of 35x35x105 mm3 . By integrating a microlens array (MLA) into the optical train of a handheld microscope, the biological specimen's image will be captured for ease of use in a single shot. With the light field raw data and program, the focal plane can be changed digitally and the 3-D image can be reconstructed after the image was taken. To localize an object in a 3-D volume, an automated data analysis algorithm to precisely distinguish profundity position is needed. The ability to create focal stacks from a single image allows moving or specimens to be recorded. Applying light field microscope algorithm to these focal stacks, a set of cross sections will be produced, which can be visualized using 3-D rendering. Furthermore, we have developed a series of design rules in order to enhance the pixel using efficiency and reduce the crosstalk between each microlens for obtain good image quality. In this paper, we demonstrate a handheld light field microscope (HLFM) to distinguish two different color fluorescence particles separated by a cover glass in a 600um range, show its focal stacks, and 3-D position.

  7. Structured light field 3D imaging.

    PubMed

    Cai, Zewei; Liu, Xiaoli; Peng, Xiang; Yin, Yongkai; Li, Ameng; Wu, Jiachen; Gao, Bruce Z

    2016-09-01

    In this paper, we propose a method by means of light field imaging under structured illumination to deal with high dynamic range 3D imaging. Fringe patterns are projected onto a scene and modulated by the scene depth then a structured light field is detected using light field recording devices. The structured light field contains information about ray direction and phase-encoded depth, via which the scene depth can be estimated from different directions. The multidirectional depth estimation can achieve high dynamic 3D imaging effectively. We analyzed and derived the phase-depth mapping in the structured light field and then proposed a flexible ray-based calibration approach to determine the independent mapping coefficients for each ray. Experimental results demonstrated the validity of the proposed method to perform high-quality 3D imaging for highly and lowly reflective surfaces. PMID:27607639

  8. 3D quantitative phase imaging of neural networks using WDT

    NASA Astrophysics Data System (ADS)

    Kim, Taewoo; Liu, S. C.; Iyer, Raj; Gillette, Martha U.; Popescu, Gabriel

    2015-03-01

    White-light diffraction tomography (WDT) is a recently developed 3D imaging technique based on a quantitative phase imaging system called spatial light interference microscopy (SLIM). The technique has achieved a sub-micron resolution in all three directions with high sensitivity granted by the low-coherence of a white-light source. Demonstrations of the technique on single cell imaging have been presented previously; however, imaging on any larger sample, including a cluster of cells, has not been demonstrated using the technique. Neurons in an animal body form a highly complex and spatially organized 3D structure, which can be characterized by neuronal networks or circuits. Currently, the most common method of studying the 3D structure of neuron networks is by using a confocal fluorescence microscope, which requires fluorescence tagging with either transient membrane dyes or after fixation of the cells. Therefore, studies on neurons are often limited to samples that are chemically treated and/or dead. WDT presents a solution for imaging live neuron networks with a high spatial and temporal resolution, because it is a 3D imaging method that is label-free and non-invasive. Using this method, a mouse or rat hippocampal neuron culture and a mouse dorsal root ganglion (DRG) neuron culture have been imaged in order to see the extension of processes between the cells in 3D. Furthermore, the tomogram is compared with a confocal fluorescence image in order to investigate the 3D structure at synapses.

  9. 3-D stimulated emission depletion microscopy with programmable aberration correction.

    PubMed

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

    2014-01-01

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

  10. ICER-3D Hyperspectral Image Compression Software

    NASA Technical Reports Server (NTRS)

    Xie, Hua; Kiely, Aaron; Klimesh, matthew; Aranki, Nazeeh

    2010-01-01

    Software has been developed to implement the ICER-3D algorithm. ICER-3D effects progressive, three-dimensional (3D), wavelet-based compression of hyperspectral images. If a compressed data stream is truncated, the progressive nature of the algorithm enables reconstruction of hyperspectral data at fidelity commensurate with the given data volume. The ICER-3D software is capable of providing either lossless or lossy compression, and incorporates an error-containment scheme to limit the effects of data loss during transmission. The compression algorithm, which was derived from the ICER image compression algorithm, includes wavelet-transform, context-modeling, and entropy coding subalgorithms. The 3D wavelet decomposition structure used by ICER-3D exploits correlations in all three dimensions of sets of hyperspectral image data, while facilitating elimination of spectral ringing artifacts, using a technique summarized in "Improving 3D Wavelet-Based Compression of Spectral Images" (NPO-41381), NASA Tech Briefs, Vol. 33, No. 3 (March 2009), page 7a. Correlation is further exploited by a context-modeling subalgorithm, which exploits spectral dependencies in the wavelet-transformed hyperspectral data, using an algorithm that is summarized in "Context Modeler for Wavelet Compression of Hyperspectral Images" (NPO-43239), which follows this article. An important feature of ICER-3D is a scheme for limiting the adverse effects of loss of data during transmission. In this scheme, as in the similar scheme used by ICER, the spatial-frequency domain is partitioned into rectangular error-containment regions. In ICER-3D, the partitions extend through all the wavelength bands. The data in each partition are compressed independently of those in the other partitions, so that loss or corruption of data from any partition does not affect the other partitions. Furthermore, because compression is progressive within each partition, when data are lost, any data from that partition received

  11. Acquisition and applications of 3D images

    NASA Astrophysics Data System (ADS)

    Sterian, Paul; Mocanu, Elena

    2007-08-01

    The moiré fringes method and their analysis up to medical and entertainment applications are discussed in this paper. We describe the procedure of capturing 3D images with an Inspeck Camera that is a real-time 3D shape acquisition system based on structured light techniques. The method is a high-resolution one. After processing the images, using computer, we can use the data for creating laser fashionable objects by engraving them with a Q-switched Nd:YAG. In medical field we mention the plastic surgery and the replacement of X-Ray especially in pediatric use.

  12. Imaging and 3D morphological analysis of collagen fibrils.

    PubMed

    Altendorf, H; Decencière, E; Jeulin, D; De sa Peixoto, P; Deniset-Besseau, A; Angelini, E; Mosser, G; Schanne-Klein, M-C

    2012-08-01

    The recent booming of multiphoton imaging of collagen fibrils by means of second harmonic generation microscopy generates the need for the development and automation of quantitative methods for image analysis. Standard approaches sequentially analyse two-dimensional (2D) slices to gain knowledge on the spatial arrangement and dimension of the fibrils, whereas the reconstructed three-dimensional (3D) image yields better information about these characteristics. In this work, a 3D analysis method is proposed for second harmonic generation images of collagen fibrils, based on a recently developed 3D fibre quantification method. This analysis uses operators from mathematical morphology. The fibril structure is scanned with a directional distance transform. Inertia moments of the directional distances yield the main fibre orientation, corresponding to the main inertia axis. The collaboration of directional distances and fibre orientation delivers a geometrical estimate of the fibre radius. The results include local maps as well as global distribution of orientation and radius of the fibrils over the 3D image. They also bring a segmentation of the image into foreground and background, as well as a classification of the foreground pixels into the preferred orientations. This accurate determination of the spatial arrangement of the fibrils within a 3D data set will be most relevant in biomedical applications. It brings the possibility to monitor remodelling of collagen tissues upon a variety of injuries and to guide tissues engineering because biomimetic 3D organizations and density are requested for better integration of implants.

  13. Automating Shallow 3D Seismic Imaging

    SciTech Connect

    Steeples, Don; Tsoflias, George

    2009-01-15

    Our efforts since 1997 have been directed toward developing ultra-shallow seismic imaging as a cost-effective method applicable to DOE facilities. This report covers the final year of grant-funded research to refine 3D shallow seismic imaging, which built on a previous 7-year grant (FG07-97ER14826) that refined and demonstrated the use of an automated method of conducting shallow seismic surveys; this represents a significant departure from conventional seismic-survey field procedures. The primary objective of this final project was to develop an automated three-dimensional (3D) shallow-seismic reflection imaging capability. This is a natural progression from our previous published work and is conceptually parallel to the innovative imaging methods used in the petroleum industry.

  14. Walker Ranch 3D seismic images

    DOE Data Explorer

    Robert J. Mellors

    2016-03-01

    Amplitude images (both vertical and depth slices) extracted from 3D seismic reflection survey over area of Walker Ranch area (adjacent to Raft River). Crossline spacing of 660 feet and inline of 165 feet using a Vibroseis source. Processing included depth migration. Micro-earthquake hypocenters on images. Stratigraphic information and nearby well tracks added to images. Images are embedded in a Microsoft Word document with additional information. Exact location and depth restricted for proprietary reasons. Data collection and processing funded by Agua Caliente. Original data remains property of Agua Caliente.

  15. Longitudinal, 3D in vivo imaging of sebaceous glands by coherent anti-Stokes Raman scattering microscopy –normal function and response to cryotherapy

    PubMed Central

    Jung, Yookyung; Tam, Joshua; Jalian, H. Ray; Anderson, R. Rox; Evans, Conor L.

    2014-01-01

    Sebaceous glands perform complex functions, and are centrally involved in the pathogenesis of acne vulgaris. Current techniques for studying sebaceous glands are mostly static in nature, whereas the gland’s main function – excretion of sebum via the holocrine mechanism – can only be evaluated over time. We present a longitudinal, real-time alternative – the in vivo, label-free imaging of sebaceous glands using Coherent Anti-Stokes Raman Scattering (CARS) microscopy, which is used to selectively visualize lipids. In mouse ears, CARS microscopy revealed dynamic changes in sebaceous glands during the holocrine secretion process, as well as in response to damage to the glands caused by cooling. Detailed gland structure, plus the active migration of individual sebocytes and cohorts of sebocytes were measured. Cooling produced characteristic changes in sebocyte structure and migration. This study demonstrates that CARS microscopy is a promising tool for studying the sebaceous gland and its associated disorders in three-dimensions in vivo. PMID:25026458

  16. 3D Micro-topography of Transferred Laboratory and Natural Ice Crystal Surfaces Imaged by Cryo and Environmental Scanning Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Magee, N. B.; Boaggio, K.; Bancroft, L.; Bandamede, M.

    2015-12-01

    Recent work has highlighted micro-scale roughness on the surfaces of ice crystals grown and imaged in-situ within the chambers of environmental scanning electron microscopes (ESEM). These observations appear to align with theoretical and satellite observations that suggest a prevalence of rough ice in cirrus clouds. However, the atmospheric application of the lab observations are indeterminate because the observations have been based only on crystals grown on substrates and in pure-water vapor environments. In this work, we present details and results from the development of a transfer technique which allows natural and lab-grown ice and snow crystals to be captured, preserved, and transferred into the ESEM for 3D imaging. Ice crystals were gathered from 1) natural snow, 2) a balloon-borne cirrus particle capture device, and 3) lab-grown ice crystals from a diffusion chamber. Ice crystals were captured in a pre-conditioned small-volume (~1 cm3) cryo-containment cell. The cell was then sealed closed and transferred to a specially-designed cryogenic dewer (filled with liquid nitrogen or crushed dry ice) for transport to a new Hitachi Field Emission, Variable Pressure SEM (SU-5000). The cryo-cell was then removed from the dewer and quickly placed onto the pre-conditioned cryo transfer stage attached to the ESEM (Quorum 3010T). Quantitative 3D topographical digital elevation models of ice surfaces are reported from SEM for the first time, including a variety of objective measures of statistical surface roughness. The surfaces of the transported crystals clearly exhibit signatures of mesoscopic roughening that are similar to examples of roughness seen in ESEM-grown crystals. For most transported crystals, the habits and crystal edges are more intricate that those observed for ice grown directly on substrates within the ESEM chamber. Portions of some crystals do appear smooth even at magnification greater than 1000x, a rare observation in our ESEM-grown crystals. The

  17. Metrological characterization of 3D imaging devices

    NASA Astrophysics Data System (ADS)

    Guidi, G.

    2013-04-01

    Manufacturers often express the performance of a 3D imaging device in various non-uniform ways for the lack of internationally recognized standard requirements for metrological parameters able to identify the capability of capturing a real scene. For this reason several national and international organizations in the last ten years have been developing protocols for verifying such performance. Ranging from VDI/VDE 2634, published by the Association of German Engineers and oriented to the world of mechanical 3D measurements (triangulation-based devices), to the ASTM technical committee E57, working also on laser systems based on direct range detection (TOF, Phase Shift, FM-CW, flash LADAR), this paper shows the state of the art about the characterization of active range devices, with special emphasis on measurement uncertainty, accuracy and resolution. Most of these protocols are based on special objects whose shape and size are certified with a known level of accuracy. By capturing the 3D shape of such objects with a range device, a comparison between the measured points and the theoretical shape they should represent is possible. The actual deviations can be directly analyzed or some derived parameters can be obtained (e.g. angles between planes, distances between barycenters of spheres rigidly connected, frequency domain parameters, etc.). This paper shows theoretical aspects and experimental results of some novel characterization methods applied to different categories of active 3D imaging devices based on both principles of triangulation and direct range detection.

  18. 3D MR imaging in real time

    NASA Astrophysics Data System (ADS)

    Guttman, Michael A.; McVeigh, Elliot R.

    2001-05-01

    A system has been developed to produce live 3D volume renderings from an MR scanner. Whereas real-time 2D MR imaging has been demonstrated by several groups, 3D volumes are currently rendered off-line to gain greater understanding of anatomical structures. For example, surgical planning is sometimes performed by viewing 2D images or 3D renderings from previously acquired image data. A disadvantage of this approach is misregistration which could occur if the anatomy changes due to normal muscle contractions or surgical manipulation. The ability to produce volume renderings in real-time and present them in the magnet room could eliminate this problem, and enable or benefit other types of interventional procedures. The system uses the data stream generated by a fast 2D multi- slice pulse sequence to update a volume rendering immediately after a new slice is available. We demonstrate some basic types of user interaction with the rendering during imaging at a rate of up to 20 frames per second.

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

    PubMed

    Nelson, A C

    1986-01-01

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

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

    PubMed

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

    2016-05-01

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

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

    PubMed

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

    2016-05-01

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

  2. Teat Morphology Characterization With 3D Imaging.

    PubMed

    Vesterinen, Heidi M; Corfe, Ian J; Sinkkonen, Ville; Iivanainen, Antti; Jernvall, Jukka; Laakkonen, Juha

    2015-07-01

    The objective of this study was to visualize, in a novel way, the morphological characteristics of bovine teats to gain a better understanding of the detailed teat morphology. We applied silicone casting and 3D digital imaging in order to obtain a more detailed image of the teat structures than that seen in previous studies. Teat samples from 65 dairy cows over 12 months of age were obtained from cows slaughtered at an abattoir. The teats were classified according to the teat condition scoring used in Finland and the lengths of the teat canals were measured. Silicone molds were made from the external teat surface surrounding the teat orifice and from the internal surface of the teat consisting of the papillary duct, Fürstenberg's rosette, and distal part of the teat cistern. The external and internal surface molds of 35 cows were scanned with a 3D laser scanner. The molds and the digital 3D models were used to evaluate internal and external teat surface morphology. A number of measurements were taken from the silicone molds. The 3D models reproduced the morphology of the teats accurately with high repeatability. Breed didn't correlate with the teat classification score. The rosette was found to have significant variation in its size and number of mucosal folds. The internal surface morphology of the rosette did not correlate with the external surface morphology of the teat implying that it is relatively independent of milking parameters that may impact the teat canal and the external surface of the teat. PMID:25382725

  3. 3D goes digital: from stereoscopy to modern 3D imaging techniques

    NASA Astrophysics Data System (ADS)

    Kerwien, N.

    2014-11-01

    In the 19th century, English physicist Charles Wheatstone discovered stereopsis, the basis for 3D perception. His construction of the first stereoscope established the foundation for stereoscopic 3D imaging. Since then, many optical instruments were influenced by these basic ideas. In recent decades, the advent of digital technologies revolutionized 3D imaging. Powerful readily available sensors and displays combined with efficient pre- or post-processing enable new methods for 3D imaging and applications. This paper draws an arc from basic concepts of 3D imaging to modern digital implementations, highlighting instructive examples from its 175 years of history.

  4. Composite model of a 3-D image

    NASA Technical Reports Server (NTRS)

    Dukhovich, I. J.

    1980-01-01

    This paper presents a composite model of a moving (3-D) image especially useful for the sequential image processing and encoding. A non-linear predictor based on the composite model is described. The performance of this predictor is used as a measure of the validity of the model for a real image source. The minimization of a total mean square prediction error provides an inequality which determines a condition for the profitable use of the composite model and can serve as a decision device for the selection of the number of subsources within the model. The paper also describes statistical properties of the prediction error and contains results of computer simulation of two non-linear predictors in the case of perfect classification between subsources.

  5. 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.

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

    PubMed

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

    2012-01-01

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

  7. [3D interactive clipping technology in medical image processing].

    PubMed

    Sun, Shaoping; Yang, Kaitai; Li, Bin; Li, Yuanjun; Liang, Jing

    2013-09-01

    The aim of this paper is to study the methods of 3D visualization and the 3D interactive clipping of CT/MRI image sequence in arbitrary orientation based on the Visualization Toolkit (VTK). A new method for 3D CT/MRI reconstructed image clipping is presented, which can clip 3D object and 3D space of medical image sequence to observe the inner structure using 3D widget for manipulating an infinite plane. Experiment results show that the proposed method can implement 3D interactive clipping of medical image effectively and get satisfied results with good quality in short time.

  8. Photogrammetric 3D reconstruction using mobile imaging

    NASA Astrophysics Data System (ADS)

    Fritsch, Dieter; Syll, Miguel

    2015-03-01

    In our paper we demonstrate the development of an Android Application (AndroidSfM) for photogrammetric 3D reconstruction that works on smartphones and tablets likewise. The photos are taken with mobile devices, and can thereafter directly be calibrated using standard calibration algorithms of photogrammetry and computer vision, on that device. Due to still limited computing resources on mobile devices, a client-server handshake using Dropbox transfers the photos to the sever to run AndroidSfM for the pose estimation of all photos by Structure-from-Motion and, thereafter, uses the oriented bunch of photos for dense point cloud estimation by dense image matching algorithms. The result is transferred back to the mobile device for visualization and ad-hoc on-screen measurements.

  9. Imaging a Sustainable Future in 3D

    NASA Astrophysics Data System (ADS)

    Schuhr, W.; Lee, J. D.; Kanngieser, E.

    2012-07-01

    It is the intention of this paper, to contribute to a sustainable future by providing objective object information based on 3D photography as well as promoting 3D photography not only for scientists, but also for amateurs. Due to the presentation of this article by CIPA Task Group 3 on "3D Photographs in Cultural Heritage", the presented samples are masterpieces of historic as well as of current 3D photography concentrating on cultural heritage. In addition to a report on exemplarily access to international archives of 3D photographs, samples for new 3D photographs taken with modern 3D cameras, as well as by means of a ground based high resolution XLITE staff camera and also 3D photographs taken from a captive balloon and the use of civil drone platforms are dealt with. To advise on optimum suited 3D methodology, as well as to catch new trends in 3D, an updated synoptic overview of the 3D visualization technology, even claiming completeness, has been carried out as a result of a systematic survey. In this respect, e.g., today's lasered crystals might be "early bird" products in 3D, which, due to lack in resolution, contrast and color, remember to the stage of the invention of photography.

  10. Ames Lab 101: Real-Time 3D Imaging

    ScienceCinema

    Zhang, Song

    2016-07-12

    Ames Laboratory scientist Song Zhang explains his real-time 3-D imaging technology. The technique can be used to create high-resolution, real-time, precise, 3-D images for use in healthcare, security, and entertainment applications.

  11. Ames Lab 101: Real-Time 3D Imaging

    SciTech Connect

    Zhang, Song

    2010-01-01

    Ames Laboratory scientist Song Zhang explains his real-time 3-D imaging technology. The technique can be used to create high-resolution, real-time, precise, 3-D images for use in healthcare, security, and entertainment applications.

  12. 3D structure of individual nanocrystals in solution by electron microscopy

    NASA Astrophysics Data System (ADS)

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

    2015-07-01

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

  13. Progress in 3D imaging and display by integral imaging

    NASA Astrophysics Data System (ADS)

    Martinez-Cuenca, R.; Saavedra, G.; Martinez-Corral, M.; Pons, A.; Javidi, B.

    2009-05-01

    Three-dimensionality is currently considered an important added value in imaging devices, and therefore the search for an optimum 3D imaging and display technique is a hot topic that is attracting important research efforts. As main value, 3D monitors should provide the observers with different perspectives of a 3D scene by simply varying the head position. Three-dimensional imaging techniques have the potential to establish a future mass-market in the fields of entertainment and communications. Integral imaging (InI), which can capture true 3D color images, has been seen as the right technology to 3D viewing to audiences of more than one person. Due to the advanced degree of development, InI technology could be ready for commercialization in the coming years. This development is the result of a strong research effort performed along the past few years by many groups. Since Integral Imaging is still an emerging technology, the first aim of the "3D Imaging and Display Laboratory" at the University of Valencia, has been the realization of a thorough study of the principles that govern its operation. Is remarkable that some of these principles have been recognized and characterized by our group. Other contributions of our research have been addressed to overcome some of the classical limitations of InI systems, like the limited depth of field (in pickup and in display), the poor axial and lateral resolution, the pseudoscopic-to-orthoscopic conversion, the production of 3D images with continuous relief, or the limited range of viewing angles of InI monitors.

  14. 3D subcellular SIMS imaging in cryogenically prepared single cells

    NASA Astrophysics Data System (ADS)

    Chandra, Subhash

    2004-06-01

    The analysis of a cell with dynamic SIMS ion microscopy depends on the gradual erosion (sputtering) of the cell surface for obtaining spatially resolved chemical information in the X-, Y-, and Z-dimensions. This ideal feature of ion microscopy is rarely explored in probing microfeatures hidden beneath the cell surface. In this study, this capability is explored for the analysis of cells undergoing cell division. The mitotic cells required 3D SIMS imaging in order to study the chemical composition of specialized subcellular regions, like the mitotic spindle, hidden beneath the cell surface. Human glioblastoma T98G cells were grown on silicon chips and cryogenically prepared with a sandwich freeze-fracture method. The fractured freeze-dried cells were used for SIMS analysis with the microscope mode of the CAMECA IMS-3f, which is capable of producing 500 nm lateral image resolution. SIMS analysis of calcium in the spindle region of metaphase cells required sequential recording of as many as 10 images. The T98G human glioblastoma tumor cells revealed an unusual depletion/lack of calcium store in the metaphase spindle, which is in contrast to the accumulation of calcium stores generally observed in normal cells. This study shows the feasibility of the microscope mode imaging in resolving subcellular microfeatures in 3D and opens new avenues of research in spatially resolved chemical analysis of dividing cells.

  15. 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.

  16. Super deep 3D images from a 3D omnifocus video camera.

    PubMed

    Iizuka, Keigo

    2012-02-20

    When using stereographic image pairs to create three-dimensional (3D) images, a deep depth of field in the original scene enhances the depth perception in the 3D image. The omnifocus video camera has no depth of field limitations and produces images that are in focus throughout. By installing an attachment on the omnifocus video camera, real-time super deep stereoscopic pairs of video images were obtained. The deeper depth of field creates a larger perspective image shift, which makes greater demands on the binocular fusion of human vision. A means of reducing the perspective shift without harming the depth of field was found.

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

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

    NASA Astrophysics Data System (ADS)

    McNally, James G.

    1994-09-01

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

  19. 3D spatial resolution and spectral resolution of interferometric 3D imaging spectrometry.

    PubMed

    Obara, Masaki; Yoshimori, Kyu

    2016-04-01

    Recently developed interferometric 3D imaging spectrometry (J. Opt. Soc. Am A18, 765 [2001]1084-7529JOAOD610.1364/JOSAA.18.000765) enables obtainment of the spectral information and 3D spatial information for incoherently illuminated or self-luminous object simultaneously. Using this method, we can obtain multispectral components of complex holograms, which correspond directly to the phase distribution of the wavefronts propagated from the polychromatic object. This paper focuses on the analysis of spectral resolution and 3D spatial resolution in interferometric 3D imaging spectrometry. Our analysis is based on a novel analytical impulse response function defined over four-dimensional space. We found that the experimental results agree well with the theoretical prediction. This work also suggests a new criterion and estimate method regarding 3D spatial resolution of digital holography. PMID:27139648

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

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed

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

    2015-11-01

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

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

    PubMed Central

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

    2015-01-01

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

  6. Automatic 2D-to-3D image conversion using 3D examples from the internet

    NASA Astrophysics Data System (ADS)

    Konrad, J.; Brown, G.; Wang, M.; Ishwar, P.; Wu, C.; Mukherjee, D.

    2012-03-01

    The availability of 3D hardware has so far outpaced the production of 3D content. Although to date many methods have been proposed to convert 2D images to 3D stereopairs, the most successful ones involve human operators and, therefore, are time-consuming and costly, while the fully-automatic ones have not yet achieved the same level of quality. This subpar performance is due to the fact that automatic methods usually rely on assumptions about the captured 3D scene that are often violated in practice. In this paper, we explore a radically different approach inspired by our work on saliency detection in images. Instead of relying on a deterministic scene model for the input 2D image, we propose to "learn" the model from a large dictionary of stereopairs, such as YouTube 3D. Our new approach is built upon a key observation and an assumption. The key observation is that among millions of stereopairs available on-line, there likely exist many stereopairs whose 3D content matches that of the 2D input (query). We assume that two stereopairs whose left images are photometrically similar are likely to have similar disparity fields. Our approach first finds a number of on-line stereopairs whose left image is a close photometric match to the 2D query and then extracts depth information from these stereopairs. Since disparities for the selected stereopairs differ due to differences in underlying image content, level of noise, distortions, etc., we combine them by using the median. We apply the resulting median disparity field to the 2D query to obtain the corresponding right image, while handling occlusions and newly-exposed areas in the usual way. We have applied our method in two scenarios. First, we used YouTube 3D videos in search of the most similar frames. Then, we repeated the experiments on a small, but carefully-selected, dictionary of stereopairs closely matching the query. This, to a degree, emulates the results one would expect from the use of an extremely large 3D

  7. Lensfree diffractive tomography for the imaging of 3D cell cultures

    PubMed Central

    Momey, F.; Berdeu, A.; Bordy, T.; Dinten, J.-M.; Marcel, F. Kermarrec; Picollet-D’hahan, N.; Gidrol, X.; Allier, C.

    2016-01-01

    New microscopes are needed to help realize the full potential of 3D organoid culture studies. In order to image large volumes of 3D organoid cultures while preserving the ability to catch every single cell, we propose a new imaging platform based on lensfree microscopy. We have built a lensfree diffractive tomography setup performing multi-angle acquisitions of 3D organoid culture embedded in Matrigel and developed a dedicated 3D holographic reconstruction algorithm based on the Fourier diffraction theorem. With this new imaging platform, we have been able to reconstruct a 3D volume as large as 21.5 mm3 of a 3D organoid culture of prostatic RWPE1 cells showing the ability of these cells to assemble in 3D intricate cellular network at the mesoscopic scale. Importantly, comparisons with 2D images show that it is possible to resolve single cells isolated from the main cellular structure with our lensfree diffractive tomography setup. PMID:27231600

  8. Lensfree diffractive tomography for the imaging of 3D cell cultures.

    PubMed

    Momey, F; Berdeu, A; Bordy, T; Dinten, J-M; Marcel, F Kermarrec; Picollet-D'hahan, N; Gidrol, X; Allier, C

    2016-03-01

    New microscopes are needed to help realize the full potential of 3D organoid culture studies. In order to image large volumes of 3D organoid cultures while preserving the ability to catch every single cell, we propose a new imaging platform based on lensfree microscopy. We have built a lensfree diffractive tomography setup performing multi-angle acquisitions of 3D organoid culture embedded in Matrigel and developed a dedicated 3D holographic reconstruction algorithm based on the Fourier diffraction theorem. With this new imaging platform, we have been able to reconstruct a 3D volume as large as 21.5 mm (3) of a 3D organoid culture of prostatic RWPE1 cells showing the ability of these cells to assemble in 3D intricate cellular network at the mesoscopic scale. Importantly, comparisons with 2D images show that it is possible to resolve single cells isolated from the main cellular structure with our lensfree diffractive tomography setup. PMID:27231600

  9. 3D seismic imaging, example of 3D area in the middle of Banat

    NASA Astrophysics Data System (ADS)

    Antic, S.

    2009-04-01

    3D seismic imaging was carried out in the 3D seismic volume situated in the middle of Banat region in Serbia. The 3D area is about 300 km square. The aim of 3D investigation was defining geology structures and techtonics especially in Mesozoik complex. The investigation objects are located in depth from 2000 to 3000 m. There are number of wells in this area but they are not enough deep to help in the interpretation. It was necessary to get better seismic image in deeper area. Acquisition parameters were satisfactory (good quality of input parameters, length of input data was 5 s, fold was up to 4000 %) and preprocessed data was satisfied. GeoDepth is an integrated system for 3D velocity model building and for 3D seismic imaging. Input data for 3D seismic imaging consist of preprocessing data sorted to CMP gathers and RMS stacking velocity functions. Other type of input data are geological information derived from well data, time migrated images and time migrated maps. Workflow for this job was: loading and quality control the input data (CMP gathers and velocity), creating initial RMS Velocity Volume, PSTM, updating the RMS Velocity Volume, PSTM, building the Initial Interval Velocity Model, PSDM, updating the Interval Velocity Model, PSDM. In the first stage the attempt is to derive initial velocity model as simple as possible as.The higher frequency velocity changes are obtained in the updating stage. The next step, after running PSTM, is the time to depth conversion. After the model is built, we generate a 3D interval velocity volume and run 3D pre-stack depth migration. The main method for updating velocities is 3D tomography. The criteria used in velocity model determination are based on the flatness of pre-stack migrated gathers or the quality of the stacked image. The standard processing ended with poststack 3D time migration. Prestack depth migration is one of the powerful tool available to the interpretator to develop an accurate velocity model and get

  10. 3D print customized sample holders for live light sheet microscopy.

    PubMed

    Jeandupeux, Emeric; Lobjois, Valérie; Ducommun, Bernard

    2015-08-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

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

  12. 3D ultrasound imaging for prosthesis fabrication and diagnostic imaging

    SciTech Connect

    Morimoto, A.K.; Bow, W.J.; Strong, D.S.

    1995-06-01

    The fabrication of a prosthetic socket for a below-the-knee amputee requires knowledge of the underlying bone structure in order to provide pressure relief for sensitive areas and support for load bearing areas. The goal is to enable the residual limb to bear pressure with greater ease and utility. Conventional methods of prosthesis fabrication are based on limited knowledge about the patient`s underlying bone structure. A 3D ultrasound imaging system was developed at Sandia National Laboratories. The imaging system provides information about the location of the bones in the residual limb along with the shape of the skin surface. Computer assisted design (CAD) software can use this data to design prosthetic sockets for amputees. Ultrasound was selected as the imaging modality. A computer model was developed to analyze the effect of the various scanning parameters and to assist in the design of the overall system. The 3D ultrasound imaging system combines off-the-shelf technology for image capturing, custom hardware, and control and image processing software to generate two types of image data -- volumetric and planar. Both volumetric and planar images reveal definition of skin and bone geometry with planar images providing details on muscle fascial planes, muscle/fat interfaces, and blood vessel definition. The 3D ultrasound imaging system was tested on 9 unilateral below-the- knee amputees. Image data was acquired from both the sound limb and the residual limb. The imaging system was operated in both volumetric and planar formats. An x-ray CT (Computed Tomography) scan was performed on each amputee for comparison. Results of the test indicate beneficial use of ultrasound to generate databases for fabrication of prostheses at a lower cost and with better initial fit as compared to manually fabricated prostheses.

  13. 3D Imaging by Mass Spectrometry: A New Frontier

    PubMed Central

    Seeley, Erin H.; Caprioli, Richard M.

    2012-01-01

    Summary Imaging mass spectrometry can generate three-dimensional volumes showing molecular distributions in an entire organ or animal through registration and stacking of serial tissue sections. Here we review the current state of 3D imaging mass spectrometry as well as provide insights and perspectives on the process of generating 3D mass spectral data along with a discussion of the process necessary to generate a 3D image volume. PMID:22276611

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

    PubMed

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

    2016-07-01

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

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

    PubMed

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

    2016-07-01

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

  16. A 3D image analysis tool for SPECT imaging

    NASA Astrophysics Data System (ADS)

    Kontos, Despina; Wang, Qiang; Megalooikonomou, Vasileios; Maurer, Alan H.; Knight, Linda C.; Kantor, Steve; Fisher, Robert S.; Simonian, Hrair P.; Parkman, Henry P.

    2005-04-01

    We have developed semi-automated and fully-automated tools for the analysis of 3D single-photon emission computed tomography (SPECT) images. The focus is on the efficient boundary delineation of complex 3D structures that enables accurate measurement of their structural and physiologic properties. We employ intensity based thresholding algorithms for interactive and semi-automated analysis. We also explore fuzzy-connectedness concepts for fully automating the segmentation process. We apply the proposed tools to SPECT image data capturing variation of gastric accommodation and emptying. These image analysis tools were developed within the framework of a noninvasive scintigraphic test to measure simultaneously both gastric emptying and gastric volume after ingestion of a solid or a liquid meal. The clinical focus of the particular analysis was to probe associations between gastric accommodation/emptying and functional dyspepsia. Employing the proposed tools, we outline effectively the complex three dimensional gastric boundaries shown in the 3D SPECT images. We also perform accurate volume calculations in order to quantitatively assess the gastric mass variation. This analysis was performed both with the semi-automated and fully-automated tools. The results were validated against manual segmentation performed by a human expert. We believe that the development of an automated segmentation tool for SPECT imaging of the gastric volume variability will allow for other new applications of SPECT imaging where there is a need to evaluate complex organ function or tumor masses.

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

    PubMed Central

    2011-01-01

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

  18. Precise 3D image alignment in micro-axial tomography.

    PubMed

    Matula, P; Kozubek, M; Staier, F; Hausmann, M

    2003-02-01

    Micro (micro-) axial tomography is a challenging technique in microscopy which improves quantitative imaging especially in cytogenetic applications by means of defined sample rotation under the microscope objective. The advantage of micro-axial tomography is an effective improvement of the precision of distance measurements between point-like objects. Under certain circumstances, the effective (3D) resolution can be improved by optimized acquisition depending on subsequent, multi-perspective image recording of the same objects followed by reconstruction methods. This requires, however, a very precise alignment of the tilted views. We present a novel feature-based image alignment method with a precision better than the full width at half maximum of the point spread function. The features are the positions (centres of gravity) of all fluorescent objects observed in the images (e.g. cell nuclei, fluorescent signals inside cell nuclei, fluorescent beads, etc.). Thus, real alignment precision depends on the localization precision of these objects. The method automatically determines the corresponding objects in subsequently tilted perspectives using a weighted bipartite graph. The optimum transformation function is computed in a least squares manner based on the coordinates of the centres of gravity of the matched objects. The theoretically feasible precision of the method was calculated using computer-generated data and confirmed by tests on real image series obtained from data sets of 200 nm fluorescent nano-particles. The advantages of the proposed algorithm are its speed and accuracy, which means that if enough objects are included, the real alignment precision is better than the axial localization precision of a single object. The alignment precision can be assessed directly from the algorithm's output. Thus, the method can be applied not only for image alignment and object matching in tilted view series in order to reconstruct (3D) images, but also to validate the

  19. 3D single molecule tracking in thick cellular specimens using multifocal plane microscopy

    NASA Astrophysics Data System (ADS)

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

    2011-03-01

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

  20. 3D motion of DNA-Au nanoconjugates in graphene liquid cell electron microscopy.

    PubMed

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

    2013-09-11

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

  1. Light field display and 3D image reconstruction

    NASA Astrophysics Data System (ADS)

    Iwane, Toru

    2016-06-01

    Light field optics and its applications become rather popular in these days. With light field optics or light field thesis, real 3D space can be described in 2D plane as 4D data, which we call as light field data. This process can be divided in two procedures. First, real3D scene is optically reduced with imaging lens. Second, this optically reduced 3D image is encoded into light field data. In later procedure we can say that 3D information is encoded onto a plane as 2D data by lens array plate. This transformation is reversible and acquired light field data can be decoded again into 3D image with the arrayed lens plate. "Refocusing" (focusing image on your favorite point after taking a picture), light-field camera's most popular function, is some kind of sectioning process from encoded 3D data (light field data) to 2D image. In this paper at first I show our actual light field camera and our 3D display using acquired and computer-simulated light field data, on which real 3D image is reconstructed. In second I explain our data processing method whose arithmetic operation is performed not in Fourier domain but in real domain. Then our 3D display system is characterized by a few features; reconstructed image is of finer resolutions than density of arrayed lenses and it is not necessary to adjust lens array plate to flat display on which light field data is displayed.

  2. Dynamic contrast-enhanced 3D photoacoustic imaging

    NASA Astrophysics Data System (ADS)

    Wong, Philip; Kosik, Ivan; Carson, Jeffrey J. L.

    2013-03-01

    Photoacoustic imaging (PAI) is a hybrid imaging modality that integrates the strengths from both optical imaging and acoustic imaging while simultaneously overcoming many of their respective weaknesses. In previous work, we reported on a real-time 3D PAI system comprised of a 32-element hemispherical array of transducers. Using the system, we demonstrated the ability to capture photoacoustic data, reconstruct a 3D photoacoustic image, and display select slices of the 3D image every 1.4 s, where each 3D image resulted from a single laser pulse. The present study aimed to exploit the rapid imaging speed of an upgraded 3D PAI system by evaluating its ability to perform dynamic contrast-enhanced imaging. The contrast dynamics can provide rich datasets that contain insight into perfusion, pharmacokinetics and physiology. We captured a series of 3D PA images of a flow phantom before and during injection of piglet and rabbit blood. Principal component analysis was utilized to classify the data according to its spatiotemporal information. The results suggested that this technique can be used to separate a sequence of 3D PA images into a series of images representative of main features according to spatiotemporal flow dynamics.

  3. 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.

  4. 3D Imaging with Structured Illumination for Advanced Security Applications

    SciTech Connect

    Birch, Gabriel Carisle; Dagel, Amber Lynn; Kast, Brian A.; Smith, Collin S.

    2015-09-01

    Three-dimensional (3D) information in a physical security system is a highly useful dis- criminator. The two-dimensional data from an imaging systems fails to provide target dis- tance and three-dimensional motion vector, which can be used to reduce nuisance alarm rates and increase system effectiveness. However, 3D imaging devices designed primarily for use in physical security systems are uncommon. This report discusses an architecture favorable to physical security systems; an inexpensive snapshot 3D imaging system utilizing a simple illumination system. The method of acquiring 3D data, tests to understand illumination de- sign, and software modifications possible to maximize information gathering capability are discussed.

  5. 3D structure tensor analysis of light microscopy data for validating diffusion MRI.

    PubMed

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

    2015-05-01

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

  6. 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

  7. On Alternative Approaches to 3D Image Perception: Monoscopic 3D Techniques

    NASA Astrophysics Data System (ADS)

    Blundell, Barry G.

    2015-06-01

    In the eighteenth century, techniques that enabled a strong sense of 3D perception to be experienced without recourse to binocular disparities (arising from the spatial separation of the eyes) underpinned the first significant commercial sales of 3D viewing devices and associated content. However following the advent of stereoscopic techniques in the nineteenth century, 3D image depiction has become inextricably linked to binocular parallax and outside the vision science and arts communities relatively little attention has been directed towards earlier approaches. Here we introduce relevant concepts and terminology and consider a number of techniques and optical devices that enable 3D perception to be experienced on the basis of planar images rendered from a single vantage point. Subsequently we allude to possible mechanisms for non-binocular parallax based 3D perception. Particular attention is given to reviewing areas likely to be thought-provoking to those involved in 3D display development, spatial visualization, HCI, and other related areas of interdisciplinary research.

  8. Advanced 3D Optical Microscopy in ENS Research.

    PubMed

    Vanden Berghe, Pieter

    2016-01-01

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

  9. Advanced 3D Optical Microscopy in ENS Research.

    PubMed

    Vanden Berghe, Pieter

    2016-01-01

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

  10. 3D augmented reality with integral imaging display

    NASA Astrophysics Data System (ADS)

    Shen, Xin; Hua, Hong; Javidi, Bahram

    2016-06-01

    In this paper, a three-dimensional (3D) integral imaging display for augmented reality is presented. By implementing the pseudoscopic-to-orthoscopic conversion method, elemental image arrays with different capturing parameters can be transferred into the identical format for 3D display. With the proposed merging algorithm, a new set of elemental images for augmented reality display is generated. The newly generated elemental images contain both the virtual objects and real world scene with desired depth information and transparency parameters. The experimental results indicate the feasibility of the proposed 3D augmented reality with integral imaging.

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

    NASA Astrophysics Data System (ADS)

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

    2003-06-01

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

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

    PubMed

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

    2015-02-28

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

  13. Object Segmentation and Ground Truth in 3D Embryonic Imaging.

    PubMed

    Rajasekaran, Bhavna; Uriu, Koichiro; Valentin, Guillaume; Tinevez, Jean-Yves; Oates, Andrew C

    2016-01-01

    Many questions in developmental biology depend on measuring the position and movement of individual cells within developing embryos. Yet, tools that provide this data are often challenged by high cell density and their accuracy is difficult to measure. Here, we present a three-step procedure to address this problem. Step one is a novel segmentation algorithm based on image derivatives that, in combination with selective post-processing, reliably and automatically segments cell nuclei from images of densely packed tissue. Step two is a quantitative validation using synthetic images to ascertain the efficiency of the algorithm with respect to signal-to-noise ratio and object density. Finally, we propose an original method to generate reliable and experimentally faithful ground truth datasets: Sparse-dense dual-labeled embryo chimeras are used to unambiguously measure segmentation errors within experimental data. Together, the three steps outlined here establish a robust, iterative procedure to fine-tune image analysis algorithms and microscopy settings associated with embryonic 3D image data sets. PMID:27332860

  14. Object Segmentation and Ground Truth in 3D Embryonic Imaging

    PubMed Central

    Rajasekaran, Bhavna; Uriu, Koichiro; Valentin, Guillaume; Tinevez, Jean-Yves; Oates, Andrew C.

    2016-01-01

    Many questions in developmental biology depend on measuring the position and movement of individual cells within developing embryos. Yet, tools that provide this data are often challenged by high cell density and their accuracy is difficult to measure. Here, we present a three-step procedure to address this problem. Step one is a novel segmentation algorithm based on image derivatives that, in combination with selective post-processing, reliably and automatically segments cell nuclei from images of densely packed tissue. Step two is a quantitative validation using synthetic images to ascertain the efficiency of the algorithm with respect to signal-to-noise ratio and object density. Finally, we propose an original method to generate reliable and experimentally faithful ground truth datasets: Sparse-dense dual-labeled embryo chimeras are used to unambiguously measure segmentation errors within experimental data. Together, the three steps outlined here establish a robust, iterative procedure to fine-tune image analysis algorithms and microscopy settings associated with embryonic 3D image data sets. PMID:27332860

  15. Dual-view 3D displays based on integral imaging

    NASA Astrophysics Data System (ADS)

    Wang, Qiong-Hua; Deng, Huan; Wu, Fei

    2016-03-01

    We propose three dual-view integral imaging (DVII) three-dimensional (3D) displays. In the spatial-multiplexed DVII 3D display, each elemental image (EI) is cut into a left and right sub-EIs, and they are refracted to the left and right viewing zones by the corresponding micro-lens array (MLA). Different 3D images are reconstructed in the left and right viewing zones, and the viewing angle is decreased. In the DVII 3D display using polarizer parallax barriers, a polarizer parallax barrier is used in front of both the display panel and the MLA. The polarizer parallax barrier consists of two parts with perpendicular polarization directions. The elemental image array (EIA) is cut to left and right parts. The lights emitted from the left part are modulated by the left MLA and reconstruct a 3D image in the right viewing zone, whereas the lights emitted from the right part reconstruct another 3D image in the left viewing zone. The 3D resolution is decreased. In the time-multiplexed DVII 3D display, an orthogonal polarizer array is attached onto both the display panel and the MLA. The orthogonal polarizer array consists of horizontal and vertical polarizer units and the polarization directions of the adjacent units are orthogonal. In State 1, each EI is reconstructed by its corresponding micro-lens, whereas in State 2, each EI is reconstructed by its adjacent micro-lens. 3D images 1 and 2 are reconstructed alternately with a refresh rate up to 120HZ. The viewing angle and 3D resolution are the same as the conventional II 3D display.

  16. 3D model-based still image object categorization

    NASA Astrophysics Data System (ADS)

    Petre, Raluca-Diana; Zaharia, Titus

    2011-09-01

    This paper proposes a novel recognition scheme algorithm for semantic labeling of 2D object present in still images. The principle consists of matching unknown 2D objects with categorized 3D models in order to infer the semantics of the 3D object to the image. We tested our new recognition framework by using the MPEG-7 and Princeton 3D model databases in order to label unknown images randomly selected from the web. Results obtained show promising performances, with recognition rate up to 84%, which opens interesting perspectives in terms of semantic metadata extraction from still images/videos.

  17. Imaging hypoxia using 3D photoacoustic spectroscopy

    NASA Astrophysics Data System (ADS)

    Stantz, Keith M.

    2010-02-01

    Purpose: The objective is to develop a multivariate in vivo hemodynamic model of tissue oxygenation (MiHMO2) based on 3D photoacoustic spectroscopy. Introduction: Low oxygen levels, or hypoxia, deprives cancer cells of oxygen and confers resistance to irradiation, some chemotherapeutic drugs, and oxygen-dependent therapies (phototherapy) leading to treatment failure and poor disease-free and overall survival. For example, clinical studies of patients with breast carcinomas, cervical cancer, and head and neck carcinomas (HNC) are more likely to suffer local reoccurrence and metastasis if their tumors are hypoxic. A novel method to non invasively measure tumor hypoxia, identify its type, and monitor its heterogeneity is devised by measuring tumor hemodynamics, MiHMO2. Material and Methods: Simulations are performed to compare tumor pO2 levels and hypoxia based on physiology - perfusion, fractional plasma volume, fractional cellular volume - and its hemoglobin status - oxygen saturation and hemoglobin concentration - based on in vivo measurements of breast, prostate, and ovarian tumors. Simulations of MiHMO2 are performed to assess the influence of scanner resolutions and different mathematic models of oxygen delivery. Results: Sensitivity of pO2 and hypoxic fraction to photoacoustic scanner resolution and dependencies on model complexity will be presented using hemodynamic parameters for different tumors. Conclusions: Photoacoustic CT spectroscopy provides a unique ability to monitor hemodynamic and cellular physiology in tissue, which can be used to longitudinally monitor tumor oxygenation and its response to anti-angiogenic therapies.

  18. Highway 3D model from image and lidar data

    NASA Astrophysics Data System (ADS)

    Chen, Jinfeng; Chu, Henry; Sun, Xiaoduan

    2014-05-01

    We present a new method of highway 3-D model construction developed based on feature extraction in highway images and LIDAR data. We describe the processing road coordinate data that connect the image frames to the coordinates of the elevation data. Image processing methods are used to extract sky, road, and ground regions as well as significant objects (such as signs and building fronts) in the roadside for the 3D model. LIDAR data are interpolated and processed to extract the road lanes as well as other features such as trees, ditches, and elevated objects to form the 3D model. 3D geometry reasoning is used to match the image features to the 3D model. Results from successive frames are integrated to improve the final model.

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

    PubMed

    Goncharsky, Alexander; Goncharsky, Anton; Durlevich, Svyatoslav

    2016-05-01

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

  20. Dedicated 3D photoacoustic breast imaging

    PubMed Central

    Kruger, Robert A.; Kuzmiak, Cherie M.; Lam, Richard B.; Reinecke, Daniel R.; Del Rio, Stephen P.; Steed, Doreen

    2013-01-01

    Purpose: To report the design and imaging methodology of a photoacoustic scanner dedicated to imaging hemoglobin distribution throughout a human breast. Methods: The authors developed a dedicated breast photoacoustic mammography (PAM) system using a spherical detector aperture based on our previous photoacoustic tomography scanner. The system uses 512 detectors with rectilinear scanning. The scan shape is a spiral pattern whose radius varies from 24 to 96 mm, thereby allowing a field of view that accommodates a wide range of breast sizes. The authors measured the contrast-to-noise ratio (CNR) using a target comprised of 1-mm dots printed on clear plastic. Each dot absorption coefficient was approximately the same as a 1-mm thickness of whole blood at 756 nm, the output wavelength of the Alexandrite laser used by this imaging system. The target was immersed in varying depths of an 8% solution of stock Liposyn II-20%, which mimics the attenuation of breast tissue (1.1 cm−1). The spatial resolution was measured using a 6 μm-diameter carbon fiber embedded in agar. The breasts of four healthy female volunteers, spanning a range of breast size from a brassiere C cup to a DD cup, were imaged using a 96-mm spiral protocol. Results: The CNR target was clearly visualized to a depth of 53 mm. Spatial resolution, which was estimated from the full width at half-maximum of a profile across the PAM image of a carbon fiber, was 0.42 mm. In the four human volunteers, the vasculature was well visualized throughout the breast tissue, including to the chest wall. Conclusions: CNR, lateral field-of-view and penetration depth of our dedicated PAM scanning system is sufficient to image breasts as large as 1335 mL, which should accommodate up to 90% of the women in the United States. PMID:24320471

  1. 3-D capacitance density imaging system

    DOEpatents

    Fasching, G.E.

    1988-03-18

    A three-dimensional capacitance density imaging of a gasified bed or the like in a containment vessel is achieved using a plurality of electrodes provided circumferentially about the bed in levels and along the bed in channels. The electrodes are individually and selectively excited electrically at each level to produce a plurality of current flux field patterns generated in the bed at each level. The current flux field patterns are suitably sensed and a density pattern of the bed at each level determined. By combining the determined density patterns at each level, a three-dimensional density image of the bed is achieved. 7 figs.

  2. 3-D seismic imaging of complex geologies

    SciTech Connect

    Womble, D.E.; Dosanjh, S.S.; VanDyke, J.P.; Oldfield, R.A.; Greenberg, D.S.

    1995-02-01

    We present three codes for the Intel Paragon that address the problem of three-dimensional seismic imaging of complex geologies. The first code models acoustic wave propagation and can be used to generate data sets to calibrate and validate seismic imaging codes. This code reported the fastest timings for acoustic wave propagation codes at a recent SEG (Society of Exploration Geophysicists) meeting. The second code implements a Kirchhoff method for pre-stack depth migration. Development of this code is almost complete, and preliminary results are presented. The third code implements a wave equation approach to seismic migration and is a Paragon implementation of a code from the ARCO Seismic Benchmark Suite.

  3. 3-D seismic imaging of complex geologies

    NASA Astrophysics Data System (ADS)

    Womble, David E.; Dosanjh, Sudip S.; Vandyke, John P.; Oldfield, Ron A.; Greenberg, David S.

    We present three codes for the Intel Paragon that address the problem of three-dimensional seismic imaging of complex geologies. The first code models acoustic wave propagation and can be used to generate data sets to calibrate and validate seismic imaging codes. This code reported the fastest timings for acoustic wave propagation codes at a recent SEG (Society of Exploration Geophysicists) meeting. The second code implements a Kirchhoff method for pre-stack depth migration. Development of this code is almost complete, and preliminary results are presented. The third code implements a wave equation approach to seismic migration and is a Paragon implementation of a code from the ARCO Seismic Benchmark Suite.

  4. High resolution 3D imaging of synchrotron generated microbeams

    SciTech Connect

    Gagliardi, Frank M.; Cornelius, Iwan; Blencowe, Anton; Franich, Rick D.; Geso, Moshi

    2015-12-15

    Purpose: Microbeam radiation therapy (MRT) techniques are under investigation at synchrotrons worldwide. Favourable outcomes from animal and cell culture studies have proven the efficacy of MRT. The aim of MRT researchers currently is to progress to human clinical trials in the near future. The purpose of this study was to demonstrate the high resolution and 3D imaging of synchrotron generated microbeams in PRESAGE® dosimeters using laser fluorescence confocal microscopy. Methods: Water equivalent PRESAGE® dosimeters were fabricated and irradiated with microbeams on the Imaging and Medical Beamline at the Australian Synchrotron. Microbeam arrays comprised of microbeams 25–50 μm wide with 200 or 400 μm peak-to-peak spacing were delivered as single, cross-fire, multidirectional, and interspersed arrays. Imaging of the dosimeters was performed using a NIKON A1 laser fluorescence confocal microscope. Results: The spatial fractionation of the MRT beams was clearly visible in 2D and up to 9 mm in depth. Individual microbeams were easily resolved with the full width at half maximum of microbeams measured on images with resolutions of as low as 0.09 μm/pixel. Profiles obtained demonstrated the change of the peak-to-valley dose ratio for interspersed MRT microbeam arrays and subtle variations in the sample positioning by the sample stage goniometer were measured. Conclusions: Laser fluorescence confocal microscopy of MRT irradiated PRESAGE® dosimeters has been validated in this study as a high resolution imaging tool for the independent spatial and geometrical verification of MRT beam delivery.

  5. Polarimetric 3D integral imaging in photon-starved conditions.

    PubMed

    Carnicer, Artur; Javidi, Bahram

    2015-03-01

    We develop a method for obtaining 3D polarimetric integral images from elemental images recorded in low light illumination conditions. Since photon-counting images are very sparse, calculation of the Stokes parameters and the degree of polarization should be handled carefully. In our approach, polarimetric 3D integral images are generated using the Maximum Likelihood Estimation and subsequently reconstructed by means of a Total Variation Denoising filter. In this way, polarimetric results are comparable to those obtained in conventional illumination conditions. We also show that polarimetric information retrieved from photon starved images can be used in 3D object recognition problems. To the best of our knowledge, this is the first report on 3D polarimetric photon counting integral imaging. PMID:25836861

  6. 3D laser imaging for concealed object identification

    NASA Astrophysics Data System (ADS)

    Berechet, Ion; Berginc, Gérard; Berechet, Stefan

    2014-09-01

    This paper deals with new optical non-conventional 3D laser imaging. Optical non-conventional imaging explores the advantages of laser imaging to form a three-dimensional image of the scene. 3D laser imaging can be used for threedimensional medical imaging, topography, surveillance, robotic vision because of ability to detect and recognize objects. In this paper, we present a 3D laser imaging for concealed object identification. The objective of this new 3D laser imaging is to provide the user a complete 3D reconstruction of the concealed object from available 2D data limited in number and with low representativeness. The 2D laser data used in this paper come from simulations that are based on the calculation of the laser interactions with the different interfaces of the scene of interest and from experimental results. We show the global 3D reconstruction procedures capable to separate objects from foliage and reconstruct a threedimensional image of the considered object. In this paper, we present examples of reconstruction and completion of three-dimensional images and we analyse the different parameters of the identification process such as resolution, the scenario of camouflage, noise impact and lacunarity degree.

  7. Phase Sensitive Cueing for 3D Objects in Overhead Images

    SciTech Connect

    Paglieroni, D

    2005-02-04

    Locating specific 3D objects in overhead images is an important problem in many remote sensing applications. 3D objects may contain either one connected component or multiple disconnected components. Solutions must accommodate images acquired with diverse sensors at various times of the day, in various seasons of the year, or under various weather conditions. Moreover, the physical manifestation of a 3D object with fixed physical dimensions in an overhead image is highly dependent on object physical dimensions, object position/orientation, image spatial resolution, and imaging geometry (e.g., obliqueness). This paper describes a two-stage computer-assisted approach for locating 3D objects in overhead images. In the matching stage, the computer matches models of 3D objects to overhead images. The strongest degree of match over all object orientations is computed at each pixel. Unambiguous local maxima in the degree of match as a function of pixel location are then found. In the cueing stage, the computer sorts image thumbnails in descending order of figure-of-merit and presents them to human analysts for visual inspection and interpretation. The figure-of-merit associated with an image thumbnail is computed from the degrees of match to a 3D object model associated with unambiguous local maxima that lie within the thumbnail. This form of computer assistance is invaluable when most of the relevant thumbnails are highly ranked, and the amount of inspection time needed is much less for the highly ranked thumbnails than for images as a whole.

  8. V3D enables real-time 3D visualization and quantitative analysis of large-scale biological image data sets.

    PubMed

    Peng, Hanchuan; Ruan, Zongcai; Long, Fuhui; Simpson, Julie H; Myers, Eugene W

    2010-04-01

    The V3D system provides three-dimensional (3D) visualization of gigabyte-sized microscopy image stacks in real time on current laptops and desktops. V3D streamlines the online analysis, measurement and proofreading of complicated image patterns by combining ergonomic functions for selecting a location in an image directly in 3D space and for displaying biological measurements, such as from fluorescent probes, using the overlaid surface objects. V3D runs on all major computer platforms and can be enhanced by software plug-ins to address specific biological problems. To demonstrate this extensibility, we built a V3D-based application, V3D-Neuron, to reconstruct complex 3D neuronal structures from high-resolution brain images. V3D-Neuron can precisely digitize the morphology of a single neuron in a fruitfly brain in minutes, with about a 17-fold improvement in reliability and tenfold savings in time compared with other neuron reconstruction tools. Using V3D-Neuron, we demonstrate the feasibility of building a 3D digital atlas of neurite tracts in the fruitfly brain. PMID:20231818

  9. 3D/3D registration of coronary CTA and biplane XA reconstructions for improved image guidance

    SciTech Connect

    Dibildox, Gerardo Baka, Nora; Walsum, Theo van; Punt, Mark; Aben, Jean-Paul; Schultz, Carl; Niessen, Wiro

    2014-09-15

    Purpose: The authors aim to improve image guidance during percutaneous coronary interventions of chronic total occlusions (CTO) by providing information obtained from computed tomography angiography (CTA) to the cardiac interventionist. To this end, the authors investigate a method to register a 3D CTA model to biplane reconstructions. Methods: The authors developed a method for registering preoperative coronary CTA with intraoperative biplane x-ray angiography (XA) images via 3D models of the coronary arteries. The models are extracted from the CTA and biplane XA images, and are temporally aligned based on CTA reconstruction phase and XA ECG signals. Rigid spatial alignment is achieved with a robust probabilistic point set registration approach using Gaussian mixture models (GMMs). This approach is extended by including orientation in the Gaussian mixtures and by weighting bifurcation points. The method is evaluated on retrospectively acquired coronary CTA datasets of 23 CTO patients for which biplane XA images are available. Results: The Gaussian mixture model approach achieved a median registration accuracy of 1.7 mm. The extended GMM approach including orientation was not significantly different (P > 0.1) but did improve robustness with regards to the initialization of the 3D models. Conclusions: The authors demonstrated that the GMM approach can effectively be applied to register CTA to biplane XA images for the purpose of improving image guidance in percutaneous coronary interventions.

  10. Critical comparison of 3D imaging approaches

    SciTech Connect

    Bennett, C L

    1999-06-03

    Currently three imaging spectrometer architectures, tunable filter, dispersive, and Fourier transform, are viable for imaging the universe in three dimensions. There are domains of greatest utility for each of these architectures. The optimum choice among the various alternative architectures is dependent on the nature of the desired observations, the maturity of the relevant technology, and the character of the backgrounds. The domain appropriate for each of the alternatives is delineated; both for instruments having ideal performance as well as for instrumentation based on currently available technology. The environment and science objectives for the Next Generation Space Telescope will be used as a specific representative case to provide a basis for comparison of the various alternatives.

  11. Acoustic 3D imaging of dental structures

    SciTech Connect

    Lewis, D.K.; Hume, W.R.; Douglass, G.D.

    1997-02-01

    Our goals for the first year of this three dimensional electodynamic imaging project was to determine how to combine flexible, individual addressable; preprocessing of array source signals; spectral extrapolation or received signals; acoustic tomography codes; and acoustic propagation modeling code. We investigated flexible, individually addressable acoustic array material to find the best match in power, sensitivity and cost and settled on PVDF sheet arrays and 3-1 composite material.

  12. Accommodation response measurements for integral 3D image

    NASA Astrophysics Data System (ADS)

    Hiura, H.; Mishina, T.; Arai, J.; Iwadate, Y.

    2014-03-01

    We measured accommodation responses under integral photography (IP), binocular stereoscopic, and real object display conditions, and viewing conditions of binocular and monocular viewing conditions. The equipment we used was an optometric device and a 3D display. We developed the 3D display for IP and binocular stereoscopic images that comprises a high-resolution liquid crystal display (LCD) and a high-density lens array. The LCD has a resolution of 468 dpi and a diagonal size of 4.8 inches. The high-density lens array comprises 106 x 69 micro lenses that have a focal length of 3 mm and diameter of 1 mm. The lenses are arranged in a honeycomb pattern. The 3D display was positioned 60 cm from an observer under IP and binocular stereoscopic display conditions. The target was presented at eight depth positions relative to the 3D display: 15, 10, and 5 cm in front of the 3D display, on the 3D display panel, and 5, 10, 15 and 30 cm behind the 3D display under the IP and binocular stereoscopic display conditions. Under the real object display condition, the target was displayed on the 3D display panel, and the 3D display was placed at the eight positions. The results suggest that the IP image induced more natural accommodation responses compared to the binocular stereoscopic image. The accommodation responses of the IP image were weaker than those of a real object; however, they showed a similar tendency with those of the real object under the two viewing conditions. Therefore, IP can induce accommodation to the depth positions of 3D images.

  13. Fast iterative image reconstruction of 3D PET data

    SciTech Connect

    Kinahan, P.E.; Townsend, D.W.; Michel, C.

    1996-12-31

    For count-limited PET imaging protocols, two different approaches to reducing statistical noise are volume, or 3D, imaging to increase sensitivity, and statistical reconstruction methods to reduce noise propagation. These two approaches have largely been developed independently, likely due to the perception of the large computational demands of iterative 3D reconstruction methods. We present results of combining the sensitivity of 3D PET imaging with the noise reduction and reconstruction speed of 2D iterative image reconstruction methods. This combination is made possible by using the recently-developed Fourier rebinning technique (FORE), which accurately and noiselessly rebins 3D PET data into a 2D data set. The resulting 2D sinograms are then reconstructed independently by the ordered-subset EM (OSEM) iterative reconstruction method, although any other 2D reconstruction algorithm could be used. We demonstrate significant improvements in image quality for whole-body 3D PET scans by using the FORE+OSEM approach compared with the standard 3D Reprojection (3DRP) algorithm. In addition, the FORE+OSEM approach involves only 2D reconstruction and it therefore requires considerably less reconstruction time than the 3DRP algorithm, or any fully 3D statistical reconstruction algorithm.

  14. Image based 3D city modeling : Comparative study

    NASA Astrophysics Data System (ADS)

    Singh, S. P.; Jain, K.; Mandla, V. R.

    2014-06-01

    3D city model is a digital representation of the Earth's surface and it's related objects such as building, tree, vegetation, and some manmade feature belonging to urban area. The demand of 3D city modeling is increasing rapidly for various engineering and non-engineering applications. Generally four main image based approaches were used for virtual 3D city models generation. In first approach, researchers were used Sketch based modeling, second method is Procedural grammar based modeling, third approach is Close range photogrammetry based modeling and fourth approach is mainly based on Computer Vision techniques. SketchUp, CityEngine, Photomodeler and Agisoft Photoscan are the main softwares to represent these approaches respectively. These softwares have different approaches & methods suitable for image based 3D city modeling. Literature study shows that till date, there is no complete such type of comparative study available to create complete 3D city model by using images. This paper gives a comparative assessment of these four image based 3D modeling approaches. This comparative study is mainly based on data acquisition methods, data processing techniques and output 3D model products. For this research work, study area is the campus of civil engineering department, Indian Institute of Technology, Roorkee (India). This 3D campus acts as a prototype for city. This study also explains various governing parameters, factors and work experiences. This research work also gives a brief introduction, strengths and weakness of these four image based techniques. Some personal comment is also given as what can do or what can't do from these softwares. At the last, this study shows; it concluded that, each and every software has some advantages and limitations. Choice of software depends on user requirements of 3D project. For normal visualization project, SketchUp software is a good option. For 3D documentation record, Photomodeler gives good result. For Large city

  15. Computational integral-imaging reconstruction-based 3-D volumetric target object recognition by using a 3-D reference object.

    PubMed

    Kim, Seung-Cheol; Park, Seok-Chan; Kim, Eun-Soo

    2009-12-01

    In this paper, we propose a novel computational integral-imaging reconstruction (CIIR)-based three-dimensional (3-D) image correlator system for the recognition of 3-D volumetric objects by employing a 3-D reference object. That is, a number of plane object images (POIs) computationally reconstructed from the 3-D reference object are used for the 3-D volumetric target recognition. In other words, simultaneous 3-D image correlations between two sets of target and reference POIs, which are depth-dependently reconstructed by using the CIIR method, are performed for effective recognition of 3-D volumetric objects in the proposed system. Successful experiments with this CIIR-based 3-D image correlator confirmed the feasibility of the proposed method.

  16. Imaging fault zones using 3D seismic image processing techniques

    NASA Astrophysics Data System (ADS)

    Iacopini, David; Butler, Rob; Purves, Steve

    2013-04-01

    Significant advances in structural analysis of deep water structure, salt tectonic and extensional rift basin come from the descriptions of fault system geometries imaged in 3D seismic data. However, even where seismic data are excellent, in most cases the trajectory of thrust faults is highly conjectural and still significant uncertainty exists as to the patterns of deformation that develop between the main faults segments, and even of the fault architectures themselves. Moreover structural interpretations that conventionally define faults by breaks and apparent offsets of seismic reflectors are commonly conditioned by a narrow range of theoretical models of fault behavior. For example, almost all interpretations of thrust geometries on seismic data rely on theoretical "end-member" behaviors where concepts as strain localization or multilayer mechanics are simply avoided. Yet analogue outcrop studies confirm that such descriptions are commonly unsatisfactory and incomplete. In order to fill these gaps and improve the 3D visualization of deformation in the subsurface, seismic attribute methods are developed here in conjunction with conventional mapping of reflector amplitudes (Marfurt & Chopra, 2007)). These signal processing techniques recently developed and applied especially by the oil industry use variations in the amplitude and phase of the seismic wavelet. These seismic attributes improve the signal interpretation and are calculated and applied to the entire 3D seismic dataset. In this contribution we will show 3D seismic examples of fault structures from gravity-driven deep-water thrust structures and extensional basin systems to indicate how 3D seismic image processing methods can not only build better the geometrical interpretations of the faults but also begin to map both strain and damage through amplitude/phase properties of the seismic signal. This is done by quantifying and delineating the short-range anomalies on the intensity of reflector amplitudes

  17. 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.

  18. Faster, higher quality volume visualization for 3D medical imaging

    NASA Astrophysics Data System (ADS)

    Kalvin, Alan D.; Laine, Andrew F.; Song, Ting

    2008-03-01

    The two major volume visualization methods used in biomedical applications are Maximum Intensity Projection (MIP) and Volume Rendering (VR), both of which involve the process of creating sets of 2D projections from 3D images. We have developed a new method for very fast, high-quality volume visualization of 3D biomedical images, based on the fact that the inverse of this process (transforming 2D projections into a 3D image) is essentially equivalent to tomographic image reconstruction. This new method uses the 2D projections acquired by the scanner, thereby obviating the need for the two computationally expensive steps currently required in the complete process of biomedical visualization, that is, (i) reconstructing the 3D image from 2D projection data, and (ii) computing the set of 2D projections from the reconstructed 3D image As well as improvements in computation speed, this method also results in improvements in visualization quality, and in the case of x-ray CT we can exploit this quality improvement to reduce radiation dosage. In this paper, demonstrate the benefits of developing biomedical visualization techniques by directly processing the sensor data acquired by body scanners, rather than by processing the image data reconstructed from the sensor data. We show results of using this approach for volume visualization for tomographic modalities, like x-ray CT, and as well as for MRI.

  19. 3D elemental sensitive imaging by full-field XFCT.

    PubMed

    Deng, Biao; Du, Guohao; Zhou, Guangzhao; Wang, Yudan; Ren, Yuqi; Chen, Rongchang; Sun, Pengfei; Xie, Honglan; Xiao, Tiqiao

    2015-05-21

    X-ray fluorescence computed tomography (XFCT) is a stimulated emission tomography modality that maps the three-dimensional (3D) distribution of elements. Generally, XFCT is done by scanning a pencil-beam across the sample. This paper presents a feasibility study of full-field XFCT (FF-XFCT) for 3D elemental imaging. The FF-XFCT consists of a pinhole collimator and X-ray imaging detector with no energy resolution. A prototype imaging system was set up at the Shanghai Synchrotron Radiation Facility (SSRF) for imaging the phantom. The first FF-XFCT experimental results are presented. The cadmium (Cd) and iodine (I) distributions were reconstructed. The results demonstrate FF-XFCT is fit for 3D elemental imaging and the sensitivity of FF-XFCT is higher than a conventional CT system.

  20. Automatic 3D lesion segmentation on breast ultrasound images

    NASA Astrophysics Data System (ADS)

    Kuo, Hsien-Chi; Giger, Maryellen L.; Reiser, Ingrid; Drukker, Karen; Edwards, Alexandra; Sennett, Charlene A.

    2013-02-01

    Automatically acquired and reconstructed 3D breast ultrasound images allow radiologists to detect and evaluate breast lesions in 3D. However, assessing potential cancers in 3D ultrasound can be difficult and time consuming. In this study, we evaluate a 3D lesion segmentation method, which we had previously developed for breast CT, and investigate its robustness on lesions on 3D breast ultrasound images. Our dataset includes 98 3D breast ultrasound images obtained on an ABUS system from 55 patients containing 64 cancers. Cancers depicted on 54 US images had been clinically interpreted as negative on screening mammography and 44 had been clinically visible on mammography. All were from women with breast density BI-RADS 3 or 4. Tumor centers and margins were indicated and outlined by radiologists. Initial RGI-eroded contours were automatically calculated and served as input to the active contour segmentation algorithm yielding the final lesion contour. Tumor segmentation was evaluated by determining the overlap ratio (OR) between computer-determined and manually-drawn outlines. Resulting average overlap ratios on coronal, transverse, and sagittal views were 0.60 +/- 0.17, 0.57 +/- 0.18, and 0.58 +/- 0.17, respectively. All OR values were significantly higher the 0.4, which is deemed "acceptable". Within the groups of mammogram-negative and mammogram-positive cancers, the overlap ratios were 0.63 +/- 0.17 and 0.56 +/- 0.16, respectively, on the coronal views; with similar results on the other views. The segmentation performance was not found to be correlated to tumor size. Results indicate robustness of the 3D lesion segmentation technique in multi-modality 3D breast imaging.

  1. 3D thermography imaging standardization technique for inflammation diagnosis

    NASA Astrophysics Data System (ADS)

    Ju, Xiangyang; Nebel, Jean-Christophe; Siebert, J. Paul

    2005-01-01

    We develop a 3D thermography imaging standardization technique to allow quantitative data analysis. Medical Digital Infrared Thermal Imaging is very sensitive and reliable mean of graphically mapping and display skin surface temperature. It allows doctors to visualise in colour and quantify temperature changes in skin surface. The spectrum of colours indicates both hot and cold responses which may co-exist if the pain associate with an inflammatory focus excites an increase in sympathetic activity. However, due to thermograph provides only qualitative diagnosis information, it has not gained acceptance in the medical and veterinary communities as a necessary or effective tool in inflammation and tumor detection. Here, our technique is based on the combination of visual 3D imaging technique and thermal imaging technique, which maps the 2D thermography images on to 3D anatomical model. Then we rectify the 3D thermogram into a view independent thermogram and conform it a standard shape template. The combination of these imaging facilities allows the generation of combined 3D and thermal data from which thermal signatures can be quantified.

  2. 3D stereophotogrammetric image superimposition onto 3D CT scan images: the future of orthognathic surgery. A pilot study.

    PubMed

    Khambay, Balvinder; Nebel, Jean-Christophe; Bowman, Janet; Walker, Fraser; Hadley, Donald M; Ayoub, Ashraf

    2002-01-01

    The aim of this study was to register and assess the accuracy of the superimposition method of a 3-dimensional (3D) soft tissue stereophotogrammetric image (C3D image) and a 3D image of the underlying skeletal tissue acquired by 3D spiral computerized tomography (CT). The study was conducted on a model head, in which an intact human skull was embedded with an overlying latex mask that reproduced anatomic features of a human face. Ten artificial radiopaque landmarks were secured to the surface of the latex mask. A stereophotogrammetric image of the mask and a 3D spiral CT image of the model head were captured. The C3D image and the CT images were registered for superimposition by 3 different methods: Procrustes superimposition using artificial landmarks, Procrustes analysis using anatomic landmarks, and partial Procrustes analysis using anatomic landmarks and then registration completion by HICP (a modified Iterative Closest Point algorithm) using a specified region of both images. The results showed that Procrustes superimposition using the artificial landmarks produced an error of superimposition on the order of 10 mm. Procrustes analysis using anatomic landmarks produced an error in the order of 2 mm. Partial Procrustes analysis using anatomic landmarks followed by HICP produced a superimposition accuracy of between 1.25 and 1.5 mm. It was concluded that a stereophotogrammetric and a 3D spiral CT scan image can be superimposed with an accuracy of between 1.25 and 1.5 mm using partial Procrustes analysis based on anatomic landmarks and then registration completion by HICP.

  3. A 3D surface imaging system for assessing human obesity

    NASA Astrophysics Data System (ADS)

    Xu, B.; Yu, W.; Yao, M.; Yao, X.; Li, Q.; Pepper, M. R.; Freeland-Graves, J. H.

    2009-08-01

    The increasing prevalence of obesity suggests a need to develop a convenient, reliable and economical tool for assessment of this condition. Three-dimensional (3D) body surface imaging has emerged as an exciting technology for estimation of body composition. This paper presents a new 3D body imaging system, which was designed for enhanced portability, affordability, and functionality. In this system, stereo vision technology was used to satisfy the requirements for a simple hardware setup and fast image acquisitions. The portability of the system was created via a two-stand configuration, and the accuracy of body volume measurements was improved by customizing stereo matching and surface reconstruction algorithms that target specific problems in 3D body imaging. Body measurement functions dedicated to body composition assessment also were developed. The overall performance of the system was evaluated in human subjects by comparison to other conventional anthropometric methods, as well as air displacement plethysmography, for body fat assessment.

  4. Hybrid segmentation framework for 3D medical image analysis

    NASA Astrophysics Data System (ADS)

    Chen, Ting; Metaxas, Dimitri N.

    2003-05-01

    Medical image segmentation is the process that defines the region of interest in the image volume. Classical segmentation methods such as region-based methods and boundary-based methods cannot make full use of the information provided by the image. In this paper we proposed a general hybrid framework for 3D medical image segmentation purposes. In our approach we combine the Gibbs Prior model, and the deformable model. First, Gibbs Prior models are applied onto each slice in a 3D medical image volume and the segmentation results are combined to a 3D binary masks of the object. Then we create a deformable mesh based on this 3D binary mask. The deformable model will be lead to the edge features in the volume with the help of image derived external forces. The deformable model segmentation result can be used to update the parameters for Gibbs Prior models. These methods will then work recursively to reach a global segmentation solution. The hybrid segmentation framework has been applied to images with the objective of lung, heart, colon, jaw, tumor, and brain. The experimental data includes MRI (T1, T2, PD), CT, X-ray, Ultra-Sound images. High quality results are achieved with relatively efficient time cost. We also did validation work using expert manual segmentation as the ground truth. The result shows that the hybrid segmentation may have further clinical use.

  5. 3D image analysis of abdominal aortic aneurysm

    NASA Astrophysics Data System (ADS)

    Subasic, Marko; Loncaric, Sven; Sorantin, Erich

    2001-07-01

    In this paper we propose a technique for 3-D segmentation of abdominal aortic aneurysm (AAA) from computed tomography angiography (CTA) images. Output data (3-D model) form the proposed method can be used for measurement of aortic shape and dimensions. Knowledge of aortic shape and size is very important in planning of minimally invasive procedure that is for selection of appropriate stent graft device for treatment of AAA. The technique is based on a 3-D deformable model and utilizes the level-set algorithm for implementation of the method. The method performs 3-D segmentation of CTA images and extracts a 3-D model of aortic wall. Once the 3-D model of aortic wall is available it is easy to perform all required measurements for appropriate stent graft selection. The method proposed in this paper uses the level-set algorithm for deformable models, instead of the classical snake algorithm. The main advantage of the level set algorithm is that it enables easy segmentation of complex structures, surpassing most of the drawbacks of the classical approach. We have extended the deformable model to incorporate the a priori knowledge about the shape of the AAA. This helps direct the evolution of the deformable model to correctly segment the aorta. The algorithm has been implemented in IDL and C languages. Experiments have been performed using real patient CTA images and have shown good results.

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

    PubMed

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

    2013-01-31

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

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

    PubMed

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

    2013-01-31

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

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

    PubMed

    Kim, Boyoung; Naemura, Takeshi

    2016-06-01

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

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

    PubMed

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

    2012-10-22

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

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

    NASA Astrophysics Data System (ADS)

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

    2010-08-01

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

  11. 3-D Terahertz Synthetic-Aperture Imaging and Spectroscopy

    NASA Astrophysics Data System (ADS)

    Henry, Samuel C.

    Terahertz (THz) wavelengths have attracted recent interest in multiple disciplines within engineering and science. Situated between the infrared and the microwave region of the electromagnetic spectrum, THz energy can propagate through non-polar materials such as clothing or packaging layers. Moreover, many chemical compounds, including explosives and many drugs, reveal strong absorption signatures in the THz range. For these reasons, THz wavelengths have great potential for non-destructive evaluation and explosive detection. Three-dimensional (3-D) reflection imaging with considerable depth resolution is also possible using pulsed THz systems. While THz imaging (especially 3-D) systems typically operate in transmission mode, reflection offers the most practical configuration for standoff detection, especially for objects with high water content (like human tissue) which are opaque at THz frequencies. In this research, reflection-based THz synthetic-aperture (SA) imaging is investigated as a potential imaging solution. THz SA imaging results presented in this dissertation are unique in that a 2-D planar synthetic array was used to generate a 3-D image without relying on a narrow time-window for depth isolation cite [Shen 2005]. Novel THz chemical detection techniques are developed and combined with broadband THz SA capabilities to provide concurrent 3-D spectral imaging. All algorithms are tested with various objects and pressed pellets using a pulsed THz time-domain system in the Northwest Electromagnetics and Acoustics Research Laboratory (NEAR-Lab).

  12. Computerized analysis of pelvic incidence from 3D images

    NASA Astrophysics Data System (ADS)

    Vrtovec, Tomaž; Janssen, Michiel M. A.; Pernuš, Franjo; Castelein, René M.; Viergever, Max A.

    2012-02-01

    The sagittal alignment of the pelvis can be evaluated by the angle of pelvic incidence (PI), which is constant for an arbitrary subject position and orientation and can be therefore compared among subjects in standing, sitting or supine position. In this study, PI was measured from three-dimensional (3D) computed tomography (CT) images of normal subjects that were acquired in supine position. A novel computerized method, based on image processing techniques, was developed to automatically determine the anatomical references required to measure PI, i.e. the centers of the femoral heads in 3D, and the center and inclination of the sacral endplate in 3D. Multiplanar image reformation was applied to obtain perfect sagittal views with all anatomical structures completely in line with the hip axis, from which PI was calculated. The resulting PI (mean+/-standard deviation) was equal to 46.6°+/-9.2° for male subjects (N = 189), 47.6°+/-10.7° for female subjects (N = 181), and 47.1°+/-10.0° for all subjects (N = 370). The obtained measurements of PI from 3D images were not biased by acquisition projection or structure orientation, because all anatomical structures were completely in line with the hip axis. The performed measurements in 3D therefore represent PI according to the actual geometrical relationships among anatomical structures of the sacrum, pelvis and hips, as observed from the perfect sagittal views.

  13. 3D image analysis of abdominal aortic aneurysm

    NASA Astrophysics Data System (ADS)

    Subasic, Marko; Loncaric, Sven; Sorantin, Erich

    2002-05-01

    This paper presents a method for 3-D segmentation of abdominal aortic aneurysm from computed tomography angiography images. The proposed method is automatic and requires minimal user assistance. Segmentation is performed in two steps. First inner and then outer aortic border is segmented. Those two steps are different due to different image conditions on two aortic borders. Outputs of these two segmentations give a complete 3-D model of abdominal aorta. Such a 3-D model is used in measurements of aneurysm area. The deformable model is implemented using the level-set algorithm due to its ability to describe complex shapes in natural manner which frequently occur in pathology. In segmentation of outer aortic boundary we introduced some knowledge based preprocessing to enhance and reconstruct low contrast aortic boundary. The method has been implemented in IDL and C languages. Experiments have been performed using real patient CTA images and have shown good results.

  14. MMSE Reconstruction for 3D Freehand Ultrasound Imaging

    PubMed Central

    Huang, Wei; Zheng, Yibin

    2008-01-01

    The reconstruction of 3D ultrasound (US) images from mechanically registered, but otherwise irregularly positioned, B-scan slices is of great interest in image guided therapy procedures. Conventional 3D ultrasound algorithms have low computational complexity, but the reconstructed volume suffers from severe speckle contamination. Furthermore, the current method cannot reconstruct uniform high-resolution data from several low-resolution B-scans. In this paper, the minimum mean-squared error (MMSE) method is applied to 3D ultrasound reconstruction. Data redundancies due to overlapping samples as well as correlation of the target and speckle are naturally accounted for in the MMSE reconstruction algorithm. Thus, the reconstruction process unifies the interpolation and spatial compounding. Simulation results for synthetic US images are presented to demonstrate the excellent reconstruction. PMID:18382623

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

    NASA Astrophysics Data System (ADS)

    Xie, Yiting; Reeves, Anthony P.

    2014-03-01

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

  16. Optimized Bayes variational regularization prior for 3D PET images.

    PubMed

    Rapisarda, Eugenio; Presotto, Luca; De Bernardi, Elisabetta; Gilardi, Maria Carla; Bettinardi, Valentino

    2014-09-01

    A new prior for variational Maximum a Posteriori regularization is proposed to be used in a 3D One-Step-Late (OSL) reconstruction algorithm accounting also for the Point Spread Function (PSF) of the PET system. The new regularization prior strongly smoothes background regions, while preserving transitions. A detectability index is proposed to optimize the prior. The new algorithm has been compared with different reconstruction algorithms such as 3D-OSEM+PSF, 3D-OSEM+PSF+post-filtering and 3D-OSL with a Gauss-Total Variation (GTV) prior. The proposed regularization allows controlling noise, while maintaining good signal recovery; compared to the other algorithms it demonstrates a very good compromise between an improved quantitation and good image quality. PMID:24958594

  17. 3D CARS image reconstruction and pattern recognition on SHG images

    NASA Astrophysics Data System (ADS)

    Medyukhina, Anna; Vogler, Nadine; Latka, Ines; Dietzek, Benjamin; Cicchi, Riccardo; Pavone, Francesco S.; Popp, Jürgen

    2012-06-01

    Nonlinear optical imaging techniques based e.g. on coherent anti-Stokes Raman scattering (CARS) or second-harmonic generation (SHG) show great potential for in-vivo investigations of tissue. While the microspectroscopic imaging tools are established, automized data evaluation, i.e. image pattern recognition and automized image classification, of nonlinear optical images still bares great possibilities for future developments towards an objective clinical diagnosis. This contribution details the capability of nonlinear microscopy for both 3D visualization of human tissues and automated discrimination between healthy and diseased patterns using ex-vivo human skin samples. By means of CARS image alignment we show how to obtain a quasi-3D model of a skin biopsy, which allows us to trace the tissue structure in different projections. Furthermore, the potential of automated pattern and organization recognition to distinguish between healthy and keloidal skin tissue is discussed. A first classification algorithm employs the intrinsic geometrical features of collagen, which can be efficiently visualized by SHG microscopy. The shape of the collagen pattern allows conclusions about the physiological state of the skin, as the typical wavy collagen structure of healthy skin is disturbed e.g. in keloid formation. Based on the different collagen patterns a quantitative score characterizing the collagen waviness - and hence reflecting the physiological state of the tissue - is obtained. Further, two additional scoring methods for collagen organization, respectively based on a statistical analysis of the mutual organization of fibers and on FFT, are presented.

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

    PubMed Central

    Torrano, Adriano A

    2014-01-01

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

  19. 3D surface reconstruction and FIB microscopy of worn alumina hip prostheses

    NASA Astrophysics Data System (ADS)

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

    2008-08-01

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

  20. Laboratory 3D Micro-XRF/Micro-CT Imaging System

    NASA Astrophysics Data System (ADS)

    Bruyndonckx, P.; Sasov, A.; Liu, X.

    2011-09-01

    A prototype micro-XRF laboratory system based on pinhole imaging was developed to produce 3D elemental maps. The fluorescence x-rays are detected by a deep-depleted CCD camera operating in photon-counting mode. A charge-clustering algorithm, together with dynamically adjusted exposure times, ensures a correct energy measurement. The XRF component has a spatial resolution of 70 μm and an energy resolution of 180 eV at 6.4 keV. The system is augmented by a micro-CT imaging modality. This is used for attenuation correction of the XRF images and to co-register features in the 3D XRF images with morphological structures visible in the volumetric CT images of the object.

  1. A miniature high resolution 3-D imaging sonar.

    PubMed

    Josserand, Tim; Wolley, Jason

    2011-04-01

    This paper discusses the design and development of a miniature, high resolution 3-D imaging sonar. The design utilizes frequency steered phased arrays (FSPA) technology. FSPAs present a small, low-power solution to the problem of underwater imaging sonars. The technology provides a method to build sonars with a large number of beams without the proportional power, circuitry and processing complexity. The design differs from previous methods in that the array elements are manufactured from a monolithic material. With this technique the arrays are flat and considerably smaller element dimensions are achievable which allows for higher frequency ranges and smaller array sizes. In the current frequency range, the demonstrated array has ultra high image resolution (1″ range×1° azimuth×1° elevation) and small size (<3″×3″). The design of the FSPA utilizes the phasing-induced frequency-dependent directionality of a linear phased array to produce multiple beams in a forward sector. The FSPA requires only two hardware channels per array and can be arranged in single and multiple array configurations that deliver wide sector 2-D images. 3-D images can be obtained by scanning the array in a direction perpendicular to the 2-D image field and applying suitable image processing to the multiple scanned 2-D images. This paper introduces the 3-D FSPA concept, theory and design methodology. Finally, results from a prototype array are presented and discussed.

  2. 3-D Display Of Magnetic Resonance Imaging Of The Spine

    NASA Astrophysics Data System (ADS)

    Nelson, Alan C.; Kim, Yongmin; Haralick, Robert M.; Anderson, Paul A.; Johnson, Roger H.; DeSoto, Larry A.

    1988-06-01

    The original data is produced through standard magnetic resonance imaging (MRI) procedures with a surface coil applied to the lower back of a normal human subject. The 3-D spine image data consists of twenty-six contiguous slices with 256 x 256 pixels per slice. Two methods for visualization of the 3-D spine are explored. One method utilizes a verifocal mirror system which creates a true 3-D virtual picture of the object. Another method uses a standard high resolution monitor to simultaneously show the three orthogonal sections which intersect at any user-selected point within the object volume. We discuss the application of these systems in assessment of low back pain.

  3. Imaging interferometric microscopy.

    PubMed

    Schwarz, Christian J; Kuznetsova, Yuliya; Brueck, S R J

    2003-08-15

    We introduce and demonstrate a new microscopy concept: imaging interferometric microscopy (IIM), which is related to holography, synthetic-aperture imaging, and off-axis-dark-field illumination techniques. IIM is a wavelength-division multiplex approach to image formation that combines multiple images covering different spatial-frequency regions to form a composite image with a resolution much greater than that permitted by the same optical system using conventional techniques. This new type of microscopy involves both off-axis coherent illumination and reinjection of appropriate zero-order reference beams. Images demonstrate high resolution, comparable with that of a high-numerical-aperture (NA) objective, while they retain the long working distance, the large depth of field, and the large field of view of a low-NA objective. A Fourier-optics model of IIM is in good agreement with the experiment. PMID:12943079

  4. Wave-CAIPI for Highly Accelerated 3D Imaging

    PubMed Central

    Bilgic, Berkin; Gagoski, Borjan A.; Cauley, Stephen F.; Fan, Audrey P.; Polimeni, Jonathan R.; Grant, P. Ellen; Wald, Lawrence L.; Setsompop, Kawin

    2014-01-01

    Purpose To introduce the Wave-CAIPI (Controlled Aliasing in Parallel Imaging) acquisition and reconstruction technique for highly accelerated 3D imaging with negligible g-factor and artifact penalties. Methods The Wave-CAIPI 3D acquisition involves playing sinusoidal gy and gz gradients during the readout of each kx encoding line, while modifying the 3D phase encoding strategy to incur inter-slice shifts as in 2D-CAIPI acquisitions. The resulting acquisition spreads the aliasing evenly in all spatial directions, thereby taking full advantage of 3D coil sensitivity distribution. By expressing the voxel spreading effect as a convolution in image space, an efficient reconstruction scheme that does not require data gridding is proposed. Rapid acquisition and high quality image reconstruction with Wave-CAIPI is demonstrated for high-resolution magnitude and phase imaging and Quantitative Susceptibility Mapping (QSM). Results Wave-CAIPI enables full-brain gradient echo (GRE) acquisition at 1 mm isotropic voxel size and R=3×3 acceleration with maximum g-factors of 1.08 at 3T, and 1.05 at 7T. Relative to the other advanced Cartesian encoding strategies 2D-CAIPI and Bunched Phase Encoding, Wave-CAIPI yields up to 2-fold reduction in maximum g-factor for 9-fold acceleration at both field strengths. Conclusion Wave-CAIPI allows highly accelerated 3D acquisitions with low artifact and negligible g-factor penalties, and may facilitate clinical application of high-resolution volumetric imaging. PMID:24986223

  5. Reduction of attenuation effects in 3D transrectal ultrasound images

    NASA Astrophysics Data System (ADS)

    Frimmel, Hans; Acosta, Oscar; Fenster, Aaron; Ourselin, Sébastien

    2007-03-01

    Ultrasound (US) is one of the most used imaging modalities today as it is cheap, reliable, safe and widely available. There are a number of issues with US images in general. Besides reflections which is the basis of ultrasonic imaging, other phenomena such as diffraction, refraction, attenuation, dispersion and scattering appear when ultrasound propagates through different tissues. The generated images are therefore corrupted by false boundaries, lack of signal for surface tangential to ultrasound propagation, large amount of noise giving rise to local properties, and anisotropic sampling space complicating image processing tasks. Although 3D Transrectal US (TRUS) probes are not yet widely available, within a few years they will likely be introduced in hospitals. Therefore, the improvement of automatic segmentation from 3D TRUS images, making the process independent of human factor is desirable. We introduce an algorithm for attenuation correction, reducing enhancement/shadowing effects and average attenuation effects in 3D US images, taking into account the physical properties of US. The parameters of acquisition such as logarithmic correction are unknown, therefore no additional information is available to restore the image. As the physical properties are related to the direction of each US ray, the 3D US data set is resampled into cylindrical coordinates using a fully automatic algorithm. Enhancement and shadowing effects, as well as average attenuation effects, are then removed with a rescaling process optimizing simultaneously in and perpendicular to the US ray direction. A set of tests using anisotropic diffusion are performed to illustrate the improvement in image quality, where well defined structures are visible. The evolution of both the entropy and the contrast show that our algorithm is a suitable pre-processing step for segmentation tasks.

  6. Automated curved planar reformation of 3D spine images

    NASA Astrophysics Data System (ADS)

    Vrtovec, Tomaz; Likar, Bostjan; Pernus, Franjo

    2005-10-01

    Traditional techniques for visualizing anatomical structures are based on planar cross-sections from volume images, such as images obtained by computed tomography (CT) or magnetic resonance imaging (MRI). However, planar cross-sections taken in the coordinate system of the 3D image often do not provide sufficient or qualitative enough diagnostic information, because planar cross-sections cannot follow curved anatomical structures (e.g. arteries, colon, spine, etc). Therefore, not all of the important details can be shown simultaneously in any planar cross-section. To overcome this problem, reformatted images in the coordinate system of the inspected structure must be created. This operation is usually referred to as curved planar reformation (CPR). In this paper we propose an automated method for CPR of 3D spine images, which is based on the image transformation from the standard image-based to a novel spine-based coordinate system. The axes of the proposed spine-based coordinate system are determined on the curve that represents the vertebral column, and the rotation of the vertebrae around the spine curve, both of which are described by polynomial models. The optimal polynomial parameters are obtained in an image analysis based optimization framework. The proposed method was qualitatively and quantitatively evaluated on five CT spine images. The method performed well on both normal and pathological cases and was consistent with manually obtained ground truth data. The proposed spine-based CPR benefits from reduced structural complexity in favour of improved feature perception of the spine. The reformatted images are diagnostically valuable and enable easier navigation, manipulation and orientation in 3D space. Moreover, reformatted images may prove useful for segmentation and other image analysis tasks.

  7. Imaging thin-bed reservoirs with 3-D seismic

    SciTech Connect

    Hardage, B.A.

    1996-12-01

    This article explains how a 3-D seismic data volume, a vertical seismic profile (VSP), electric well logs and reservoir pressure data can be used to image closely stacked thin-bed reservoirs. This interpretation focuses on the Oligocene Frio reservoir in South Texas which has multiple thin-beds spanning a vertical interval of about 3,000 ft.

  8. 3D imaging lidar for lunar robotic exploration

    NASA Astrophysics Data System (ADS)

    Hussein, Marwan W.; Tripp, Jeffrey W.

    2009-05-01

    Part of the requirements of the future Constellation program is to optimize lunar surface operations and reduce hazards to astronauts. Toward this end, many robotic platforms, rovers in specific, are being sought to carry out a multitude of missions involving potential EVA sites survey, surface reconnaissance, path planning and obstacle detection and classification. 3D imaging lidar technology provides an enabling capability that allows fast, accurate and detailed collection of three-dimensional information about the rover's environment. The lidar images the region of interest by scanning a laser beam and measuring the pulse time-of-flight and the bearing. The accumulated set of laser ranges and bearings constitutes the threedimensional image. As part of the ongoing NASA Ames research center activities in lunar robotics, the utility of 3D imaging lidar was evaluated by testing Optech's ILRIS-3D lidar on board the K-10 Red rover during the recent Human - Robotics Systems (HRS) field trails in Lake Moses, WA. This paper examines the results of the ILRIS-3D trials, presents the data obtained and discusses its application in lunar surface robotic surveying and scouting.

  9. Hollow Cone Electron Imaging for Single Particle 3D Reconstruction of Proteins.

    PubMed

    Tsai, Chun-Ying; Chang, Yuan-Chih; Lobato, Ivan; Van Dyck, Dirk; Chen, Fu-Rong

    2016-01-01

    The main bottlenecks for high-resolution biological imaging in electron microscopy are radiation sensitivity and low contrast. The phase contrast at low spatial frequencies can be enhanced by using a large defocus but this strongly reduces the resolution. Recently, phase plates have been developed to enhance the contrast at small defocus but electrical charging remains a problem. Single particle cryo-electron microscopy is mostly used to minimize the radiation damage and to enhance the resolution of the 3D reconstructions but it requires averaging images of a massive number of individual particles. Here we present a new route to achieve the same goals by hollow cone dark field imaging using thermal diffuse scattered electrons giving about a 4 times contrast increase as compared to bright field imaging. We demonstrate the 3D reconstruction of a stained GroEL particle can yield about 13.5 Å resolution but using a strongly reduced number of images. PMID:27292544

  10. Hollow Cone Electron Imaging for Single Particle 3D Reconstruction of Proteins

    PubMed Central

    Tsai, Chun-Ying; Chang, Yuan-Chih; Lobato, Ivan; Van Dyck, Dirk; Chen, Fu-Rong

    2016-01-01

    The main bottlenecks for high-resolution biological imaging in electron microscopy are radiation sensitivity and low contrast. The phase contrast at low spatial frequencies can be enhanced by using a large defocus but this strongly reduces the resolution. Recently, phase plates have been developed to enhance the contrast at small defocus but electrical charging remains a problem. Single particle cryo-electron microscopy is mostly used to minimize the radiation damage and to enhance the resolution of the 3D reconstructions but it requires averaging images of a massive number of individual particles. Here we present a new route to achieve the same goals by hollow cone dark field imaging using thermal diffuse scattered electrons giving about a 4 times contrast increase as compared to bright field imaging. We demonstrate the 3D reconstruction of a stained GroEL particle can yield about 13.5 Å resolution but using a strongly reduced number of images. PMID:27292544

  11. Hollow Cone Electron Imaging for Single Particle 3D Reconstruction of Proteins

    NASA Astrophysics Data System (ADS)

    Tsai, Chun-Ying; Chang, Yuan-Chih; Lobato, Ivan; van Dyck, Dirk; Chen, Fu-Rong

    2016-06-01

    The main bottlenecks for high-resolution biological imaging in electron microscopy are radiation sensitivity and low contrast. The phase contrast at low spatial frequencies can be enhanced by using a large defocus but this strongly reduces the resolution. Recently, phase plates have been developed to enhance the contrast at small defocus but electrical charging remains a problem. Single particle cryo-electron microscopy is mostly used to minimize the radiation damage and to enhance the resolution of the 3D reconstructions but it requires averaging images of a massive number of individual particles. Here we present a new route to achieve the same goals by hollow cone dark field imaging using thermal diffuse scattered electrons giving about a 4 times contrast increase as compared to bright field imaging. We demonstrate the 3D reconstruction of a stained GroEL particle can yield about 13.5 Å resolution but using a strongly reduced number of images.

  12. Practical pseudo-3D registration for large tomographic images

    NASA Astrophysics Data System (ADS)

    Liu, Xuan; Laperre, Kjell; Sasov, Alexander

    2014-09-01

    Image registration is a powerful tool in various tomographic applications. Our main focus is on microCT applications in which samples/animals can be scanned multiple times under different conditions or at different time points. For this purpose, a registration tool capable of handling fairly large volumes has been developed, using a novel pseudo-3D method to achieve fast and interactive registration with simultaneous 3D visualization. To reduce computation complexity in 3D registration, we decompose it into several 2D registrations, which are applied to the orthogonal views (transaxial, sagittal and coronal) sequentially and iteratively. After registration in each view, the next view is retrieved with the new transformation matrix for registration. This reduces the computation complexity significantly. For rigid transform, we only need to search for 3 parameters (2 shifts, 1 rotation) in each of the 3 orthogonal views instead of 6 (3 shifts, 3 rotations) for full 3D volume. In addition, the amount of voxels involved is also significantly reduced. For the proposed pseudo-3D method, image-based registration is employed, with Sum of Square Difference (SSD) as the similarity measure. The searching engine is Powell's conjugate direction method. In this paper, only rigid transform is used. However, it can be extended to affine transform by adding scaling and possibly shearing to the transform model. We have noticed that more information can be used in the 2D registration if Maximum Intensity Projections (MIP) or Parallel Projections (PP) is used instead of the orthogonal views. Also, other similarity measures, such as covariance or mutual information, can be easily incorporated. The initial evaluation on microCT data shows very promising results. Two application examples are shown: dental samples before and after treatment and structural changes in materials before and after compression. Evaluation on registration accuracy between pseudo-3D method and true 3D method has

  13. 3D wavefront image formation for NIITEK GPR

    NASA Astrophysics Data System (ADS)

    Soumekh, Mehrdad; Ton, Tuan; Howard, Pete

    2009-05-01

    The U.S. Department of Defense Humanitarian Demining (HD) Research and Development Program focuses on developing, testing, demonstrating, and validating new technology for immediate use in humanitarian demining operations around the globe. Beginning in the late 1990's, the U.S. Army Countermine Division funded the development of the NIITEK ground penetrating radar (GPR) for detection of anti-tank (AT) landmines. This work is concerned with signal processing algorithms to suppress sources of artifacts in the NIITEK GPR, and formation of three-dimensional (3D) imagery from the resultant data. We first show that the NIITEK GPR data correspond to a 3D Synthetic Aperture Radar (SAR) database. An adaptive filtering method is utilized to suppress ground return and self-induced resonance (SIR) signals that are generated by the interaction of the radar-carrying platform and the transmitted radar signal. We examine signal processing methods to improve the fidelity of imagery for this 3D SAR system using pre-processing methods that suppress Doppler aliasing as well as other side lobe leakage artifacts that are introduced by the radar radiation pattern. The algorithm, known as digital spotlighting, imposes a filtering scheme on the azimuth-compressed SAR data, and manipulates the resultant spectral data to achieve a higher PRF to suppress the Doppler aliasing. We also present the 3D version of the Fourier-based wavefront reconstruction, a computationally-efficient and approximation-free SAR imaging method, for image formation with the NIITEK 3D SAR database.

  14. Optimizing 3D image quality and performance for stereoscopic gaming

    NASA Astrophysics Data System (ADS)

    Flack, Julien; Sanderson, Hugh; Pegg, Steven; Kwok, Simon; Paterson, Daniel

    2009-02-01

    The successful introduction of stereoscopic TV systems, such as Samsung's 3D Ready Plasma, requires high quality 3D content to be commercially available to the consumer. Console and PC games provide the most readily accessible source of high quality 3D content. This paper describes innovative developments in a generic, PC-based game driver architecture that addresses the two key issues affecting 3D gaming: quality and speed. At the heart of the quality issue are the same considerations that studios face producing stereoscopic renders from CG movies: how best to perform the mapping from a geometric CG environment into the stereoscopic display volume. The major difference being that for game drivers this mapping cannot be choreographed by hand but must be automatically calculated in real-time without significant impact on performance. Performance is a critical issue when dealing with gaming. Stereoscopic gaming has traditionally meant rendering the scene twice with the associated performance overhead. An alternative approach is to render the scene from one virtual camera position and use information from the z-buffer to generate a stereo pair using Depth-Image-Based Rendering (DIBR). We analyze this trade-off in more detail and provide some results relating to both 3D image quality and render performance.

  15. 3D Winding Number: Theory and Application to Medical Imaging

    PubMed Central

    Becciu, Alessandro; Fuster, Andrea; Pottek, Mark; van den Heuvel, Bart; ter Haar Romeny, Bart; van Assen, Hans

    2011-01-01

    We develop a new formulation, mathematically elegant, to detect critical points of 3D scalar images. It is based on a topological number, which is the generalization to three dimensions of the 2D winding number. We illustrate our method by considering three different biomedical applications, namely, detection and counting of ovarian follicles and neuronal cells and estimation of cardiac motion from tagged MR images. Qualitative and quantitative evaluation emphasizes the reliability of the results. PMID:21317978

  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. 2D/3D image (facial) comparison using camera matching.

    PubMed

    Goos, Mirelle I M; Alberink, Ivo B; Ruifrok, Arnout C C

    2006-11-10

    A problem in forensic facial comparison of images of perpetrators and suspects is that distances between fixed anatomical points in the face, which form a good starting point for objective, anthropometric comparison, vary strongly according to the position and orientation of the camera. In case of a cooperating suspect, a 3D image may be taken using e.g. a laser scanning device. By projecting the 3D image onto a 2D image with the suspect's head in the same pose as that of the perpetrator, using the same focal length and pixel aspect ratio, numerical comparison of (ratios of) distances between fixed points becomes feasible. An experiment was performed in which, starting from two 3D scans and one 2D image of two colleagues, male and female, and using seven fixed anatomical locations in the face, comparisons were made for the matching and non-matching case. Using this method, the non-matching pair cannot be distinguished from the matching pair of faces. Facial expression and resolution of images were all more or less optimal, and the results of the study are not encouraging for the use of anthropometric arguments in the identification process. More research needs to be done though on larger sets of facial comparisons. PMID:16337353

  18. An automated 3D reconstruction method of UAV images

    NASA Astrophysics Data System (ADS)

    Liu, Jun; Wang, He; Liu, Xiaoyang; Li, Feng; Sun, Guangtong; Song, Ping

    2015-10-01

    In this paper a novel fully automated 3D reconstruction approach based on low-altitude unmanned aerial vehicle system (UAVs) images will be presented, which does not require previous camera calibration or any other external prior knowledge. Dense 3D point clouds are generated by integrating orderly feature extraction, image matching, structure from motion (SfM) and multi-view stereo (MVS) algorithms, overcoming many of the cost, time limitations of rigorous photogrammetry techniques. An image topology analysis strategy is introduced to speed up large scene reconstruction by taking advantage of the flight-control data acquired by UAV. Image topology map can significantly reduce the running time of feature matching by limiting the combination of images. A high-resolution digital surface model of the study area is produced base on UAV point clouds by constructing the triangular irregular network. Experimental results show that the proposed approach is robust and feasible for automatic 3D reconstruction of low-altitude UAV images, and has great potential for the acquisition of spatial information at large scales mapping, especially suitable for rapid response and precise modelling in disaster emergency.

  19. Preliminary comparison of 3D synthetic aperture imaging with Explososcan

    NASA Astrophysics Data System (ADS)

    Rasmussen, Morten Fischer; Hansen, Jens Munk; Férin, Guillaume; Dufait, Rémi; Jensen, Jørgen Arendt

    2012-03-01

    Explososcan is the 'gold standard' for real-time 3D medical ultrasound imaging. In this paper, 3D synthetic aperture imaging is compared to Explososcan by simulation of 3D point spread functions. The simulations mimic a 32×32 element prototype transducer. The transducer mimicked is a dense matrix phased array with a pitch of 300 μm, made by Vermon. For both imaging techniques, 289 emissions are used to image a volume spanning 60° in both the azimuth and elevation direction and 150mm in depth. This results for both techniques in a frame rate of 18 Hz. The implemented synthetic aperture technique reduces the number of transmit channels from 1024 to 256, compared to Explososcan. In terms of FWHM performance, was Explososcan and synthetic aperture found to perform similar. At 90mm depth is Explososcan's FWHM performance 7% better than that of synthetic aperture. Synthetic aperture improved the cystic resolution, which expresses the ability to detect anechoic cysts in a uniform scattering media, at all depths except at Explososcan's focus point. Synthetic aperture reduced the cyst radius, R20dB, at 90mm depth by 48%. Synthetic aperture imaging was shown to reduce the number of transmit channels by four and still, generally, improve the imaging quality.

  20. Refraction Correction in 3D Transcranial Ultrasound Imaging

    PubMed Central

    Lindsey, Brooks D.; Smith, Stephen W.

    2014-01-01

    We present the first correction of refraction in three-dimensional (3D) ultrasound imaging using an iterative approach that traces propagation paths through a two-layer planar tissue model, applying Snell’s law in 3D. This approach is applied to real-time 3D transcranial ultrasound imaging by precomputing delays offline for several skull thicknesses, allowing the user to switch between three sets of delays for phased array imaging at the push of a button. Simulations indicate that refraction correction may be expected to increase sensitivity, reduce beam steering errors, and partially restore lost spatial resolution, with the greatest improvements occurring at the largest steering angles. Distorted images of cylindrical lesions were created by imaging through an acrylic plate in a tissue-mimicking phantom. As a result of correcting for refraction, lesions were restored to 93.6% of their original diameter in the lateral direction and 98.1% of their original shape along the long axis of the cylinders. In imaging two healthy volunteers, the mean brightness increased by 8.3% and showed no spatial dependency. PMID:24275538

  1. Label free cell tracking in 3D tissue engineering constructs with high resolution imaging

    NASA Astrophysics Data System (ADS)

    Smith, W. A.; Lam, K.-P.; Dempsey, K. P.; Mazzocchi-Jones, D.; Richardson, J. B.; Yang, Y.

    2014-02-01

    Within the field of tissue engineering there is an emphasis on studying 3-D live tissue structures. Consequently, to investigate and identify cellular activities and phenotypes in a 3-D environment for all in vitro experiments, including shape, migration/proliferation and axon projection, it is necessary to adopt an optical imaging system that enables monitoring 3-D cellular activities and morphology through the thickness of the construct for an extended culture period without cell labeling. This paper describes a new 3-D tracking algorithm developed for Cell-IQ®, an automated cell imaging platform, which has been equipped with an environmental chamber optimized to enable capturing time-lapse sequences of live cell images over a long-term period without cell labeling. As an integral part of the algorithm, a novel auto-focusing procedure was developed for phase contrast microscopy equipped with 20x and 40x objectives, to provide a more accurate estimation of cell growth/trajectories by allowing 3-D voxels to be computed at high spatiotemporal resolution and cell density. A pilot study was carried out in a phantom system consisting of horizontally aligned nanofiber layers (with precise spacing between them), to mimic features well exemplified in cellular activities of neuronal growth in a 3-D environment. This was followed by detailed investigations concerning axonal projections and dendritic circuitry formation in a 3-D tissue engineering construct. Preliminary work on primary animal neuronal cells in response to chemoattractant and topographic cue within the scaffolds has produced encouraging results.

  2. 1024 pixels single photon imaging array for 3D ranging

    NASA Astrophysics Data System (ADS)

    Bellisai, S.; Guerrieri, F.; Tisa, S.; Zappa, F.; Tosi, A.; Giudice, A.

    2011-01-01

    Three dimensions (3D) acquisition systems are driving applications in many research field. Nowadays 3D acquiring systems are used in a lot of applications, such as cinema industry or in automotive (for active security systems). Depending on the application, systems present different features, for example color sensitivity, bi-dimensional image resolution, distance measurement accuracy and acquisition frame rate. The system we developed acquires 3D movie using indirect Time of Flight (iTOF), starting from phase delay measurement of a sinusoidally modulated light. The system acquires live movie with a frame rate up to 50frame/s in a range distance between 10 cm up to 7.5 m.

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

    NASA Astrophysics Data System (ADS)

    Oldham, Mark; Kim, Leonard; Hugo, Geoffrey

    2005-04-01

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

  4. 3D robust digital image correlation for vibration measurement.

    PubMed

    Chen, Zhong; Zhang, Xianmin; Fatikow, Sergej

    2016-03-01

    Discrepancies of speckle images under dynamic measurement due to the different viewing angles will deteriorate the correspondence in 3D digital image correlation (3D-DIC) for vibration measurement. Facing this kind of bottleneck, this paper presents two types of robust 3D-DIC methods for vibration measurement, SSD-robust and SWD-robust, which use a sum of square difference (SSD) estimator plus a Geman-McClure regulating term and a Welch estimator plus a Geman-McClure regulating term, respectively. Because the regulating term with an adaptive rejecting bound can lessen the influence of the abnormal pixel data in the dynamical measuring process, the robustness of the algorithm is enhanced. The robustness and precision evaluation experiments using a dual-frequency laser interferometer are implemented. The experimental results indicate that the two presented robust estimators can suppress the effects of the abnormality in the speckle images and, meanwhile, keep higher precision in vibration measurement in contrast with the traditional SSD method; thus, the SWD-robust and SSD-robust methods are suitable for weak image noise and strong image noise, respectively. PMID:26974624

  5. 3D Reconstruction of Human Motion from Monocular Image Sequences.

    PubMed

    Wandt, Bastian; Ackermann, Hanno; Rosenhahn, Bodo

    2016-08-01

    This article tackles the problem of estimating non-rigid human 3D shape and motion from image sequences taken by uncalibrated cameras. Similar to other state-of-the-art solutions we factorize 2D observations in camera parameters, base poses and mixing coefficients. Existing methods require sufficient camera motion during the sequence to achieve a correct 3D reconstruction. To obtain convincing 3D reconstructions from arbitrary camera motion, our method is based on a-priorly trained base poses. We show that strong periodic assumptions on the coefficients can be used to define an efficient and accurate algorithm for estimating periodic motion such as walking patterns. For the extension to non-periodic motion we propose a novel regularization term based on temporal bone length constancy. In contrast to other works, the proposed method does not use a predefined skeleton or anthropometric constraints and can handle arbitrary camera motion. We achieve convincing 3D reconstructions, even under the influence of noise and occlusions. Multiple experiments based on a 3D error metric demonstrate the stability of the proposed method. Compared to other state-of-the-art methods our algorithm shows a significant improvement. PMID:27093439

  6. 3D Reconstruction of Human Motion from Monocular Image Sequences.

    PubMed

    Wandt, Bastian; Ackermann, Hanno; Rosenhahn, Bodo

    2016-08-01

    This article tackles the problem of estimating non-rigid human 3D shape and motion from image sequences taken by uncalibrated cameras. Similar to other state-of-the-art solutions we factorize 2D observations in camera parameters, base poses and mixing coefficients. Existing methods require sufficient camera motion during the sequence to achieve a correct 3D reconstruction. To obtain convincing 3D reconstructions from arbitrary camera motion, our method is based on a-priorly trained base poses. We show that strong periodic assumptions on the coefficients can be used to define an efficient and accurate algorithm for estimating periodic motion such as walking patterns. For the extension to non-periodic motion we propose a novel regularization term based on temporal bone length constancy. In contrast to other works, the proposed method does not use a predefined skeleton or anthropometric constraints and can handle arbitrary camera motion. We achieve convincing 3D reconstructions, even under the influence of noise and occlusions. Multiple experiments based on a 3D error metric demonstrate the stability of the proposed method. Compared to other state-of-the-art methods our algorithm shows a significant improvement.

  7. Extraction of 3D information from sonar image sequences.

    PubMed

    Trucco, A; Curletto, S

    2003-01-01

    This paper describes a set of methods that make it possible to estimate the position of a feature inside a three-dimensional (3D) space by starting from a sequence of two-dimensional (2D) acoustic images of the seafloor acquired with a sonar system. Typical sonar imaging systems are able to generate just 2D images, and the acquisition of 3D information involves sharp increases in complexity and costs. The front-scan sonar proposed in this paper is a new equipment devoted to acquiring a 2D image of the seafloor to sail over, and allows one to collect a sequence of images showing a specific feature during the approach of the ship. This fact seems to make it possible to recover the 3D position of a feature by comparing the feature positions along the sequence of images acquired from different (known) ship positions. This opportunity is investigated in the paper, where it is shown that encouraging results have been obtained by a processing chain composed of some blocks devoted to low-level processing, feature extraction and analysis, a Kalman filter for robust feature tracking, and some ad hoc equations for depth estimation and averaging. A statistical error analysis demonstrated the great potential of the proposed system also if some inaccuracies affect the sonar measures and the knowledge of the ship position. This was also confirmed by several tests performed on both simulated and real sequences, obtaining satisfactory results on both the feature tracking and, above all, the estimation of the 3D position.

  8. Quantitative 3-D imaging topogrammetry for telemedicine applications

    NASA Technical Reports Server (NTRS)

    Altschuler, Bruce R.

    1994-01-01

    The technology to reliably transmit high-resolution visual imagery over short to medium distances in real time has led to the serious considerations of the use of telemedicine, telepresence, and telerobotics in the delivery of health care. These concepts may involve, and evolve toward: consultation from remote expert teaching centers; diagnosis; triage; real-time remote advice to the surgeon; and real-time remote surgical instrument manipulation (telerobotics with virtual reality). Further extrapolation leads to teledesign and telereplication of spare surgical parts through quantitative teleimaging of 3-D surfaces tied to CAD/CAM devices and an artificially intelligent archival data base of 'normal' shapes. The ability to generate 'topogrames' or 3-D surface numerical tables of coordinate values capable of creating computer-generated virtual holographic-like displays, machine part replication, and statistical diagnostic shape assessment is critical to the progression of telemedicine. Any virtual reality simulation will remain in 'video-game' realm until realistic dimensional and spatial relational inputs from real measurements in vivo during surgeries are added to an ever-growing statistical data archive. The challenges of managing and interpreting this 3-D data base, which would include radiographic and surface quantitative data, are considerable. As technology drives toward dynamic and continuous 3-D surface measurements, presenting millions of X, Y, Z data points per second of flexing, stretching, moving human organs, the knowledge base and interpretive capabilities of 'brilliant robots' to work as a surgeon's tireless assistants becomes imaginable. The brilliant robot would 'see' what the surgeon sees--and more, for the robot could quantify its 3-D sensing and would 'see' in a wider spectral range than humans, and could zoom its 'eyes' from the macro world to long-distance microscopy. Unerring robot hands could rapidly perform machine-aided suturing with

  9. Interactive 2D to 3D stereoscopic image synthesis

    NASA Astrophysics Data System (ADS)

    Feldman, Mark H.; Lipton, Lenny

    2005-03-01

    Advances in stereoscopic display technologies, graphic card devices, and digital imaging algorithms have opened up new possibilities in synthesizing stereoscopic images. The power of today"s DirectX/OpenGL optimized graphics cards together with adapting new and creative imaging tools found in software products such as Adobe Photoshop, provide a powerful environment for converting planar drawings and photographs into stereoscopic images. The basis for such a creative process is the focus of this paper. This article presents a novel technique, which uses advanced imaging features and custom Windows-based software that utilizes the Direct X 9 API to provide the user with an interactive stereo image synthesizer. By creating an accurate and interactive world scene with moveable and flexible depth map altered textured surfaces, perspective stereoscopic cameras with both visible frustums and zero parallax planes, a user can precisely model a virtual three-dimensional representation of a real-world scene. Current versions of Adobe Photoshop provide a creative user with a rich assortment of tools needed to highlight elements of a 2D image, simulate hidden areas, and creatively shape them for a 3D scene representation. The technique described has been implemented as a Photoshop plug-in and thus allows for a seamless transition of these 2D image elements into 3D surfaces, which are subsequently rendered to create stereoscopic views.

  10. Large distance 3D imaging of hidden objects

    NASA Astrophysics Data System (ADS)

    Rozban, Daniel; Aharon Akram, Avihai; Kopeika, N. S.; Abramovich, A.; Levanon, Assaf

    2014-06-01

    Imaging systems in millimeter waves are required for applications in medicine, communications, homeland security, and space technology. This is because there is no known ionization hazard for biological tissue, and atmospheric attenuation in this range of the spectrum is low compared to that of infrared and optical rays. The lack of an inexpensive room temperature detector makes it difficult to give a suitable real time implement for the above applications. A 3D MMW imaging system based on chirp radar was studied previously using a scanning imaging system of a single detector. The system presented here proposes to employ a chirp radar method with Glow Discharge Detector (GDD) Focal Plane Array (FPA of plasma based detectors) using heterodyne detection. The intensity at each pixel in the GDD FPA yields the usual 2D image. The value of the I-F frequency yields the range information at each pixel. This will enable 3D MMW imaging. In this work we experimentally demonstrate the feasibility of implementing an imaging system based on radar principles and FPA of inexpensive detectors. This imaging system is shown to be capable of imaging objects from distances of at least 10 meters.

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

    PubMed

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

    2016-02-01

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

  12. 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.

  13. 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.

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

    PubMed Central

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

    2015-01-01

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

  15. 3D imaging of fetus vertebra by synchrotron radiation microtomography

    NASA Astrophysics Data System (ADS)

    Peyrin, Francoise; Pateyron-Salome, Murielle; Denis, Frederic; Braillon, Pierre; Laval-Jeantet, Anne-Marie; Cloetens, Peter

    1997-10-01

    A synchrotron radiation computed microtomography system allowing high resolution 3D imaging of bone samples has been developed at ESRF. The system uses a high resolution 2D detector based on a CCd camera coupled to a fluorescent screen through light optics. The spatial resolution of the device is particularly well adapted to the imaging of bone structure. In view of studying growth, vertebra samples of fetus with differential gestational ages were imaged. The first results show that fetus vertebra is quite different from adult bone both in terms of density and organization.

  16. Advanced 3D imaging lidar concepts for long range sensing

    NASA Astrophysics Data System (ADS)

    Gordon, K. J.; Hiskett, P. A.; Lamb, R. A.

    2014-06-01

    Recent developments in 3D imaging lidar are presented. Long range 3D imaging using photon counting is now a possibility, offering a low-cost approach to integrated remote sensing with step changing advantages in size, weight and power compared to conventional analogue active imaging technology. We report results using a Geiger-mode array for time-of-flight, single photon counting lidar for depth profiling and determination of the shape and size of tree canopies and distributed surface reflections at a range of 9km, with 4μJ pulses with a frame rate of 100kHz using a low-cost fibre laser operating at a wavelength of λ=1.5 μm. The range resolution is less than 4cm providing very high depth resolution for target identification. This specification opens up several additional functionalities for advanced lidar, for example: absolute rangefinding and depth profiling for long range identification, optical communications, turbulence sensing and time-of-flight spectroscopy. Future concepts for 3D time-of-flight polarimetric and multispectral imaging lidar, with optical communications in a single integrated system are also proposed.

  17. Linear tracking for 3-D medical ultrasound imaging.

    PubMed

    Huang, Qing-Hua; Yang, Zhao; Hu, Wei; Jin, Lian-Wen; Wei, Gang; Li, Xuelong

    2013-12-01

    As the clinical application grows, there is a rapid technical development of 3-D ultrasound imaging. Compared with 2-D ultrasound imaging, 3-D ultrasound imaging can provide improved qualitative and quantitative information for various clinical applications. In this paper, we proposed a novel tracking method for a freehand 3-D ultrasound imaging system with improved portability, reduced degree of freedom, and cost. We designed a sliding track with a linear position sensor attached, and it transmitted positional data via a wireless communication module based on Bluetooth, resulting in a wireless spatial tracking modality. A traditional 2-D ultrasound probe fixed to the position sensor on the sliding track was used to obtain real-time B-scans, and the positions of the B-scans were simultaneously acquired when moving the probe along the track in a freehand manner. In the experiments, the proposed method was applied to ultrasound phantoms and real human tissues. The results demonstrated that the new system outperformed a previously developed freehand system based on a traditional six-degree-of-freedom spatial sensor in phantom and in vivo studies, indicating its merit in clinical applications for human tissues and organs. PMID:23757592

  18. 3D imaging: how to achieve highest accuracy

    NASA Astrophysics Data System (ADS)

    Luhmann, Thomas

    2011-07-01

    The generation of 3D information from images is a key technology in many different areas, e.g. in 3D modeling and representation of architectural or heritage objects, in human body motion tracking and scanning, in 3D scene analysis of traffic scenes, in industrial applications and many more. The basic concepts rely on mathematical representations of central perspective viewing as they are widely known from photogrammetry or computer vision approaches. The objectives of these methods differ, more or less, from high precision and well-structured measurements in (industrial) photogrammetry to fully-automated non-structured applications in computer vision. Accuracy and precision is a critical issue for the 3D measurement of industrial, engineering or medical objects. As state of the art, photogrammetric multi-view measurements achieve relative precisions in the order of 1:100000 to 1:200000, and relative accuracies with respect to retraceable lengths in the order of 1:50000 to 1:100000 of the largest object diameter. In order to obtain these figures a number of influencing parameters have to be optimized. These are, besides others: physical representation of object surface (targets, texture), illumination and light sources, imaging sensors, cameras and lenses, calibration strategies (camera model), orientation strategies (bundle adjustment), image processing of homologue features (target measurement, stereo and multi-image matching), representation of object or workpiece coordinate systems and object scale. The paper discusses the above mentioned parameters and offers strategies for obtaining highest accuracy in object space. Practical examples of high-quality stereo camera measurements and multi-image applications are used to prove the relevance of high accuracy in different applications, ranging from medical navigation to static and dynamic industrial measurements. In addition, standards for accuracy verifications are presented and demonstrated by practical examples

  19. Fourier plane imaging microscopy

    SciTech Connect

    Dominguez, Daniel Peralta, Luis Grave de; Alharbi, Nouf; Alhusain, Mdhaoui; Bernussi, Ayrton A.

    2014-09-14

    We show how the image of an unresolved photonic crystal can be reconstructed using a single Fourier plane (FP) image obtained with a second camera that was added to a traditional compound microscope. We discuss how Fourier plane imaging microscopy is an application of a remarkable property of the obtained FP images: they contain more information about the photonic crystals than the images recorded by the camera commonly placed at the real plane of the microscope. We argue that the experimental results support the hypothesis that surface waves, contributing to enhanced resolution abilities, were optically excited in the studied photonic crystals.

  20. Method for extracting the aorta from 3D CT images

    NASA Astrophysics Data System (ADS)

    Taeprasartsit, Pinyo; Higgins, William E.

    2007-03-01

    Bronchoscopic biopsy of the central-chest lymph nodes is vital in the staging of lung cancer. Three-dimensional multi-detector CT (MDCT) images provide vivid anatomical detail for planning bronchoscopy. Unfortunately, many lymph nodes are situated close to the aorta, and an inadvertent needle biopsy could puncture the aorta, causing serious harm. As an eventual aid for more complete planning of lymph-node biopsy, it is important to define the aorta. This paper proposes a method for extracting the aorta from a 3D MDCT chest image. The method has two main phases: (1) Off-line Model Construction, which provides a set of training cases for fitting new images, and (2) On-Line Aorta Construction, which is used for new incoming 3D MDCT images. Off-Line Model Construction is done once using several representative human MDCT images and consists of the following steps: construct a likelihood image, select control points of the medial axis of the aortic arch, and recompute the control points to obtain a constant-interval medial-axis model. On-Line Aorta Construction consists of the following operations: construct a likelihood image, perform global fitting of the precomputed models to the current case's likelihood image to find the best fitting model, perform local fitting to adjust the medial axis to local data variations, and employ a region recovery method to arrive at the complete constructed 3D aorta. The region recovery method consists of two steps: model-based and region-growing steps. This region growing method can recover regions outside the model coverage and non-circular tube structures. In our experiments, we used three models and achieved satisfactory results on twelve of thirteen test cases.

  1. 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.

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

    PubMed

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

    2015-02-01

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

  3. Phantom image results of an optimized full 3D USCT

    NASA Astrophysics Data System (ADS)

    Ruiter, Nicole V.; Zapf, Michael; Hopp, Torsten; Dapp, Robin; Gemmeke, Hartmut

    2012-03-01

    A promising candidate for improved imaging of breast cancer is ultrasound computer tomography (USCT). Current experimental USCT systems are still focused in elevation dimension resulting in a large slice thickness, limited depth of field, loss of out-of-plane reflections, and a large number of movement steps to acquire a stack of images. 3DUSCT emitting and receiving spherical wave fronts overcomes these limitations. We built an optimized 3DUSCT with nearly isotropic 3DPSF, realizing for the first time the full benefits of a 3Dsystem. In this paper results of the 3D point spread function measured with a dedicated phantom and images acquired with a clinical breast phantom are presented. The point spread function could be shown to be nearly isotropic in 3D, to have very low spatial variability and fit the predicted values. The contrast of the phantom images is very satisfactory in spite of imaging with a sparse aperture. The resolution and imaged details of the reflectivity reconstruction are comparable to a 3TeslaMRI volume of the breast phantom. Image quality and resolution is isotropic in all three dimensions, confirming the successful optimization experimentally.

  4. Combined elasticity and 3D imaging of the prostate

    NASA Astrophysics Data System (ADS)

    Li, Yinbo; Hossack, John A.

    2005-04-01

    A method is described for repeatably assessing elasticity and 3D extent of suspected prostate cancers. Elasticity is measured by controlled water inflation of a sheath placed over a modified transrectal ultrasound transducer. The benefit of using fluid inflation is that it should be possible to make repeatable, accurate, measurements of elasticity that are of interest in the serial assessment of prostate cancer progression or remission. The second aspect of the work uses auxiliary tracking arrays placed at each end of the central imaging array that allow the transducer to be rotated while simultaneously collected 'tracking' information thus allowing the position of successive image planes to be located with approximately 11% volumetric accuracy in 3D space. In this way, we present a technique for quantifying volumetric extent of suspected cancer in addition to making measures of elastic anomalies.

  5. 3D reconstruction of concave surfaces using polarisation imaging

    NASA Astrophysics Data System (ADS)

    Sohaib, A.; Farooq, A. R.; Ahmed, J.; Smith, L. N.; Smith, M. L.

    2015-06-01

    This paper presents a novel algorithm for improved shape recovery using polarisation-based photometric stereo. The majority of previous research using photometric stereo involves 3D reconstruction using both the diffuse and specular components of light; however, this paper suggests the use of the specular component only as it is the only form of light that comes directly off the surface without subsurface scattering or interreflections. Experiments were carried out on both real and synthetic surfaces. Real images were obtained using a polarisation-based photometric stereo device while synthetic images were generated using PovRay® software. The results clearly demonstrate that the proposed method can extract three-dimensional (3D) surface information effectively even for concave surfaces with complex texture and surface reflectance.

  6. Getting in touch--3D printing in forensic imaging.

    PubMed

    Ebert, Lars Chr; Thali, Michael J; Ross, Steffen

    2011-09-10

    With the increasing use of medical imaging in forensics, as well as the technological advances in rapid prototyping, we suggest combining these techniques to generate displays of forensic findings. We used computed tomography (CT), CT angiography, magnetic resonance imaging (MRI) and surface scanning with photogrammetry in conjunction with segmentation techniques to generate 3D polygon meshes. Based on these data sets, a 3D printer created colored models of the anatomical structures. Using this technique, we could create models of bone fractures, vessels, cardiac infarctions, ruptured organs as well as bitemark wounds. The final models are anatomically accurate, fully colored representations of bones, vessels and soft tissue, and they demonstrate radiologically visible pathologies. The models are more easily understood by laypersons than volume rendering or 2D reconstructions. Therefore, they are suitable for presentations in courtrooms and for educational purposes. PMID:21602004

  7. 3D scene reconstruction based on 3D laser point cloud combining UAV images

    NASA Astrophysics Data System (ADS)

    Liu, Huiyun; Yan, Yangyang; Zhang, Xitong; Wu, Zhenzhen

    2016-03-01

    It is a big challenge capturing and modeling 3D information of the built environment. A number of techniques and technologies are now in use. These include GPS, and photogrammetric application and also remote sensing applications. The experiment uses multi-source data fusion technology for 3D scene reconstruction based on the principle of 3D laser scanning technology, which uses the laser point cloud data as the basis and Digital Ortho-photo Map as an auxiliary, uses 3DsMAX software as a basic tool for building three-dimensional scene reconstruction. The article includes data acquisition, data preprocessing, 3D scene construction. The results show that the 3D scene has better truthfulness, and the accuracy of the scene meet the need of 3D scene construction.

  8. Automated Recognition of 3D Features in GPIR Images

    NASA Technical Reports Server (NTRS)

    Park, Han; Stough, Timothy; Fijany, Amir

    2007-01-01

    A method of automated recognition of three-dimensional (3D) features in images generated by ground-penetrating imaging radar (GPIR) is undergoing development. GPIR 3D images can be analyzed to detect and identify such subsurface features as pipes and other utility conduits. Until now, much of the analysis of GPIR images has been performed manually by expert operators who must visually identify and track each feature. The present method is intended to satisfy a need for more efficient and accurate analysis by means of algorithms that can automatically identify and track subsurface features, with minimal supervision by human operators. In this method, data from multiple sources (for example, data on different features extracted by different algorithms) are fused together for identifying subsurface objects. The algorithms of this method can be classified in several different ways. In one classification, the algorithms fall into three classes: (1) image-processing algorithms, (2) feature- extraction algorithms, and (3) a multiaxis data-fusion/pattern-recognition algorithm that includes a combination of machine-learning, pattern-recognition, and object-linking algorithms. The image-processing class includes preprocessing algorithms for reducing noise and enhancing target features for pattern recognition. The feature-extraction algorithms operate on preprocessed data to extract such specific features in images as two-dimensional (2D) slices of a pipe. Then the multiaxis data-fusion/ pattern-recognition algorithm identifies, classifies, and reconstructs 3D objects from the extracted features. In this process, multiple 2D features extracted by use of different algorithms and representing views along different directions are used to identify and reconstruct 3D objects. In object linking, which is an essential part of this process, features identified in successive 2D slices and located within a threshold radius of identical features in adjacent slices are linked in a

  9. Dynamic 3D computed tomography scanner for vascular imaging

    NASA Astrophysics Data System (ADS)

    Lee, Mark K.; Holdsworth, David W.; Fenster, Aaron

    2000-04-01

    A 3D dynamic computed-tomography (CT) scanner was developed for imaging objects undergoing periodic motion. The scanner system has high spatial and sufficient temporal resolution to produce quantitative tomographic/volume images of objects such as excised arterial samples perfused under physiological pressure conditions and enables the measurements of the local dynamic elastic modulus (Edyn) of the arteries in the axial and longitudinal directions. The system was comprised of a high resolution modified x-ray image intensifier (XRII) based computed tomographic system and a computer-controlled cardiac flow simulator. A standard NTSC CCD camera with a macro lens was coupled to the electro-optically zoomed XRII to acquire dynamic volumetric images. Through prospective cardiac gating and computer synchronized control, a time-resolved sequence of 20 mm thick high resolution volume images of porcine aortic specimens during one simulated cardiac cycle were obtained. Performance evaluation of the scanners illustrated that tomographic images can be obtained with resolution as high as 3.2 mm-1 with only a 9% decrease in the resolution for objects moving at velocities of 1 cm/s in 2D mode and static spatial resolution of 3.55 mm-1 with only a 14% decrease in the resolution in 3D mode for objects moving at a velocity of 10 cm/s. Application of the system for imaging of intact excised arterial specimens under simulated physiological flow/pressure conditions enabled measurements of the Edyn of the arteries with a precision of +/- kPa for the 3D scanner. Evaluation of the Edyn in the axial and longitudinal direction produced values of 428 +/- 35 kPa and 728 +/- 71 kPa, demonstrating the isotropic and homogeneous viscoelastic nature of the vascular specimens. These values obtained from the Dynamic CT systems were not statistically different (p less than 0.05) from the values obtained by standard uniaxial tensile testing and volumetric measurements.

  10. 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

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

    PubMed

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

    2016-07-01

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

  12. High-speed 3D imaging by DMD technology

    NASA Astrophysics Data System (ADS)

    Hoefling, Roland

    2004-05-01

    The paper presents an advanced solution for capturing the height of an object in addition to the 2D image as it is frequently desired in machine vision applications. Based upon the active fringe projection methodology, the system takes advantage of a series of patterns projected onto the object surface and observed by a camera to provide reliable, accurate and highly resolved 3D data from any scattering object surface. The paper shows how the recording of a projected image series can be significantly accelerated and improved in quality to overcome current limitations. The key is ALP - a metrology dedicated hardware design using the Discovery 1100 platform for the DMD micromirror device of Texas Instruments Inc. The paper describes how this DMD technology has been combined with latest LED illumination, high-performance optics, and recent digital camera solutions. The ALP based DMD projection can be exactly synchronized with one or multiple cameras so that gray value intensities generated by pulse-width modulation (PWM) are recorded with high linearity. Based upon these components, a novel 3D measuring system with outstanding properties is described. The "z-Snapper" represents a new class of 3D imaging devices, it is fast enough for time demanding in-line testing, and it can be built completely mobile: laptop based, hand-held, and battery powered. The turnkey system provides a "3D image" as simple as an usual b/w picture is grabbed. It can be instantly implemented into future machine vision applications that will benefit from the step into the third dimension.

  13. Discrete Method of Images for 3D Radio Propagation Modeling

    NASA Astrophysics Data System (ADS)

    Novak, Roman

    2016-09-01

    Discretization by rasterization is introduced into the method of images (MI) in the context of 3D deterministic radio propagation modeling as a way to exploit spatial coherence of electromagnetic propagation for fine-grained parallelism. Traditional algebraic treatment of bounding regions and surfaces is replaced by computer graphics rendering of 3D reflections and double refractions while building the image tree. The visibility of reception points and surfaces is also resolved by shader programs. The proposed rasterization is shown to be of comparable run time to that of the fundamentally parallel shooting and bouncing rays. The rasterization does not affect the signal evaluation backtracking step, thus preserving its advantage over the brute force ray-tracing methods in terms of accuracy. Moreover, the rendering resolution may be scaled back for a given level of scenario detail with only marginal impact on the image tree size. This allows selection of scene optimized execution parameters for faster execution, giving the method a competitive edge. The proposed variant of MI can be run on any GPU that supports real-time 3D graphics.

  14. Fast 3D fluid registration of brain magnetic resonance images

    NASA Astrophysics Data System (ADS)

    Leporé, Natasha; Chou, Yi-Yu; Lopez, Oscar L.; Aizenstein, Howard J.; Becker, James T.; Toga, Arthur W.; Thompson, Paul M.

    2008-03-01

    Fluid registration is widely used in medical imaging to track anatomical changes, to correct image distortions, and to integrate multi-modality data. Fluid mappings guarantee that the template image deforms smoothly into the target, without tearing or folding, even when large deformations are required for accurate matching. Here we implemented an intensity-based fluid registration algorithm, accelerated by using a filter designed by Bro-Nielsen and Gramkow. We validated the algorithm on 2D and 3D geometric phantoms using the mean square difference between the final registered image and target as a measure of the accuracy of the registration. In tests on phantom images with different levels of overlap, varying amounts of Gaussian noise, and different intensity gradients, the fluid method outperformed a more commonly used elastic registration method, both in terms of accuracy and in avoiding topological errors during deformation. We also studied the effect of varying the viscosity coefficients in the viscous fluid equation, to optimize registration accuracy. Finally, we applied the fluid registration algorithm to a dataset of 2D binary corpus callosum images and 3D volumetric brain MRIs from 14 healthy individuals to assess its accuracy and robustness.

  15. Stereotactic mammography imaging combined with 3D US imaging for image guided breast biopsy

    SciTech Connect

    Surry, K. J. M.; Mills, G. R.; Bevan, K.; Downey, D. B.; Fenster, A.

    2007-11-15

    Stereotactic X-ray mammography (SM) and ultrasound (US) guidance are both commonly used for breast biopsy. While SM provides three-dimensional (3D) targeting information and US provides real-time guidance, both have limitations. SM is a long and uncomfortable procedure and the US guided procedure is inherently two dimensional (2D), requiring a skilled physician for both safety and accuracy. The authors developed a 3D US-guided biopsy system to be integrated with, and to supplement SM imaging. Their goal is to be able to biopsy a larger percentage of suspicious masses using US, by clarifying ambiguous structures with SM imaging. Features from SM and US guided biopsy were combined, including breast stabilization, a confined needle trajectory, and dual modality imaging. The 3D US guided biopsy system uses a 7.5 MHz breast probe and is mounted on an upright SM machine for preprocedural imaging. Intraprocedural targeting and guidance was achieved with real-time 2D and near real-time 3D US imaging. Postbiopsy 3D US imaging allowed for confirmation that the needle was penetrating the target. The authors evaluated 3D US-guided biopsy accuracy of their system using test phantoms. To use mammographic imaging information, they registered the SM and 3D US coordinate systems. The 3D positions of targets identified in the SM images were determined with a target localization error (TLE) of 0.49 mm. The z component (x-ray tube to image) of the TLE dominated with a TLE{sub z} of 0.47 mm. The SM system was then registered to 3D US, with a fiducial registration error (FRE) and target registration error (TRE) of 0.82 and 0.92 mm, respectively. Analysis of the FRE and TRE components showed that these errors were dominated by inaccuracies in the z component with a FRE{sub z} of 0.76 mm and a TRE{sub z} of 0.85 mm. A stereotactic mammography and 3D US guided breast biopsy system should include breast compression for stability and safety and dual modality imaging for target localization

  16. Femoroacetabular impingement with chronic acetabular rim fracture - 3D computed tomography, 3D magnetic resonance imaging and arthroscopic correlation

    PubMed Central

    Chhabra, Avneesh; Nordeck, Shaun; Wadhwa, Vibhor; Madhavapeddi, Sai; Robertson, William J

    2015-01-01

    Femoroacetabular impingement is uncommonly associated with a large rim fragment of bone along the superolateral acetabulum. We report an unusual case of femoroacetabular impingement (FAI) with chronic acetabular rim fracture. Radiographic, 3D computed tomography, 3D magnetic resonance imaging and arthroscopy correlation is presented with discussion of relative advantages and disadvantages of various modalities in the context of FAI. PMID:26191497

  17. Pavement cracking measurements using 3D laser-scan images

    NASA Astrophysics Data System (ADS)

    Ouyang, W.; Xu, B.

    2013-10-01

    Pavement condition surveying is vital for pavement maintenance programs that ensure ride quality and traffic safety. This paper first introduces an automated pavement inspection system which uses a three-dimensional (3D) camera and a structured laser light to acquire dense transverse profiles of a pavement lane surface when it carries a moving vehicle. After the calibration, the 3D system can yield a depth resolution of 0.5 mm and a transverse resolution of 1.56 mm pixel-1 at 1.4 m camera height from the ground. The scanning rate of the camera can be set to its maximum at 5000 lines s-1, allowing the density of scanned profiles to vary with the vehicle's speed. The paper then illustrates the algorithms that utilize 3D information to detect pavement distress, such as transverse, longitudinal and alligator cracking, and presents the field tests on the system's repeatability when scanning a sample pavement in multiple runs at the same vehicle speed, at different vehicle speeds and under different weather conditions. The results show that this dedicated 3D system can capture accurate pavement images that detail surface distress, and obtain consistent crack measurements in repeated tests and under different driving and lighting conditions.

  18. Objective breast symmetry evaluation using 3-D surface imaging.

    PubMed

    Eder, Maximilian; Waldenfels, Fee V; Swobodnik, Alexandra; Klöppel, Markus; Pape, Ann-Kathrin; Schuster, Tibor; Raith, Stefan; Kitzler, Elena; Papadopulos, Nikolaos A; Machens, Hans-Günther; Kovacs, Laszlo

    2012-04-01

    This study develops an objective breast symmetry evaluation using 3-D surface imaging (Konica-Minolta V910(®) scanner) by superimposing the mirrored left breast over the right and objectively determining the mean 3-D contour difference between the 2 breast surfaces. 3 observers analyzed the evaluation protocol precision using 2 dummy models (n = 60), 10 test subjects (n = 300), clinically tested it on 30 patients (n = 900) and compared it to established 2-D measurements on 23 breast reconstructive patients using the BCCT.core software (n = 690). Mean 3-D evaluation precision, expressed as the coefficient of variation (VC), was 3.54 ± 0.18 for all human subjects without significant intra- and inter-observer differences (p > 0.05). The 3-D breast symmetry evaluation is observer independent, significantly more precise (p < 0.001) than the BCCT.core software (VC = 6.92 ± 0.88) and may play a part in an objective surgical outcome analysis after incorporation into clinical practice.

  19. 3D thermal medical image visualization tool: Integration between MRI and thermographic images.

    PubMed

    Abreu de Souza, Mauren; Chagas Paz, André Augusto; Sanches, Ionildo Jóse; Nohama, Percy; Gamba, Humberto Remigio

    2014-01-01

    Three-dimensional medical image reconstruction using different images modalities require registration techniques that are, in general, based on the stacking of 2D MRI/CT images slices. In this way, the integration of two different imaging modalities: anatomical (MRI/CT) and physiological information (infrared image), to generate a 3D thermal model, is a new methodology still under development. This paper presents a 3D THERMO interface that provides flexibility for the 3D visualization: it incorporates the DICOM parameters; different color scale palettes at the final 3D model; 3D visualization at different planes of sections; and a filtering option that provides better image visualization. To summarize, the 3D thermographc medical image visualization provides a realistic and precise medical tool. The merging of two different imaging modalities allows better quality and more fidelity, especially for medical applications in which the temperature changes are clinically significant.

  20. 3D imaging of soil pore network: two different approaches

    NASA Astrophysics Data System (ADS)

    Matrecano, M.; Di Matteo, B.; Mele, G.; Terribile, F.

    2009-04-01

    Pore geometry imaging and its quantitative description is a key factor for advances in the knowledge of physical, chemical and biological soil processes. For many years photos from flattened surfaces of undisturbed soil samples impregnated with fluorescent resin and from soil thin sections under microscope have been the only way available for exploring pore architecture at different scales. Earlier 3D representations of the internal structure of the soil based on not destructive methods have been obtained using medical tomographic systems (NMR and X-ray CT). However, images provided using such equipments, show strong limitations in terms of spatial resolution. In the last decade very good results have then been obtained using imaging from very expensive systems based on synchrotron radiation. More recently, X-ray Micro-Tomography has resulted the most widely applied being the technique showing the best compromise between costs, resolution and size of the images. Conversely, the conceptually simpler but destructive method of "serial sectioning" has been progressively neglected for technical problems in sample preparation and time consumption needed to obtain an adequate number of serial sections for correct 3D reconstruction of soil pore geometry. In this work a comparison between the two methods above has been carried out in order to define advantages, shortcomings and to point out their different potential. A cylindrical undisturbed soil sample 6.5cm in diameter and 6.5cm height of an Ap horizon of an alluvial soil showing vertic characteristics, has been reconstructed using both a desktop X-ray micro-tomograph Skyscan 1172 and the new automatic serial sectioning system SSAT (Sequential Section Automatic Tomography) set up at CNR ISAFOM in Ercolano (Italy) with the aim to overcome most of the typical limitations of such a technique. Image best resolution of 7.5 µm per voxel resulted using X-ray Micro CT while 20 µm was the best value using the serial sectioning

  1. Automatic structural matching of 3D image data

    NASA Astrophysics Data System (ADS)

    Ponomarev, Svjatoslav; Lutsiv, Vadim; Malyshev, Igor

    2015-10-01

    A new image matching technique is described. It is implemented as an object-independent hierarchical structural juxtaposition algorithm based on an alphabet of simple object-independent contour structural elements. The structural matching applied implements an optimized method of walking through a truncated tree of all possible juxtapositions of two sets of structural elements. The algorithm was initially developed for dealing with 2D images such as the aerospace photographs, and it turned out to be sufficiently robust and reliable for matching successfully the pictures of natural landscapes taken in differing seasons from differing aspect angles by differing sensors (the visible optical, IR, and SAR pictures, as well as the depth maps and geographical vector-type maps). At present (in the reported version), the algorithm is enhanced based on additional use of information on third spatial coordinates of observed points of object surfaces. Thus, it is now capable of matching the images of 3D scenes in the tasks of automatic navigation of extremely low flying unmanned vehicles or autonomous terrestrial robots. The basic principles of 3D structural description and matching of images are described, and the examples of image matching are presented.

  2. Underwater 3d Modeling: Image Enhancement and Point Cloud Filtering

    NASA Astrophysics Data System (ADS)

    Sarakinou, I.; Papadimitriou, K.; Georgoula, O.; Patias, P.

    2016-06-01

    This paper examines the results of image enhancement and point cloud filtering on the visual and geometric quality of 3D models for the representation of underwater features. Specifically it evaluates the combination of effects from the manual editing of images' radiometry (captured at shallow depths) and the selection of parameters for point cloud definition and mesh building (processed in 3D modeling software). Such datasets, are usually collected by divers, handled by scientists and used for geovisualization purposes. In the presented study, have been created 3D models from three sets of images (seafloor, part of a wreck and a small boat's wreck) captured at three different depths (3.5m, 10m and 14m respectively). Four models have been created from the first dataset (seafloor) in order to evaluate the results from the application of image enhancement techniques and point cloud filtering. The main process for this preliminary study included a) the definition of parameters for the point cloud filtering and the creation of a reference model, b) the radiometric editing of images, followed by the creation of three improved models and c) the assessment of results by comparing the visual and the geometric quality of improved models versus the reference one. Finally, the selected technique is tested on two other data sets in order to examine its appropriateness for different depths (at 10m and 14m) and different objects (part of a wreck and a small boat's wreck) in the context of an ongoing research in the Laboratory of Photogrammetry and Remote Sensing.

  3. 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.

  4. Sparse aperture 3D passive image sensing and recognition

    NASA Astrophysics Data System (ADS)

    Daneshpanah, Mehdi

    The way we perceive, capture, store, communicate and visualize the world has greatly changed in the past century Novel three dimensional (3D) imaging and display systems are being pursued both in academic and industrial settings. In many cases, these systems have revolutionized traditional approaches and/or enabled new technologies in other disciplines including medical imaging and diagnostics, industrial metrology, entertainment, robotics as well as defense and security. In this dissertation, we focus on novel aspects of sparse aperture multi-view imaging systems and their application in quantum-limited object recognition in two separate parts. In the first part, two concepts are proposed. First a solution is presented that involves a generalized framework for 3D imaging using randomly distributed sparse apertures. Second, a method is suggested to extract the profile of objects in the scene through statistical properties of the reconstructed light field. In both cases, experimental results are presented that demonstrate the feasibility of the techniques. In the second part, the application of 3D imaging systems in sensing and recognition of objects is addressed. In particular, we focus on the scenario in which only 10s of photons reach the sensor from the object of interest, as opposed to hundreds of billions of photons in normal imaging conditions. At this level, the quantum limited behavior of light will dominate and traditional object recognition practices may fail. We suggest a likelihood based object recognition framework that incorporates the physics of sensing at quantum-limited conditions. Sensor dark noise has been modeled and taken into account. This framework is applied to 3D sensing of thermal objects using visible spectrum detectors. Thermal objects as cold as 250K are shown to provide enough signature photons to be sensed and recognized within background and dark noise with mature, visible band, image forming optics and detector arrays. The results

  5. 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.

  6. 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

  7. Feature detection on 3D images of dental imprints

    NASA Astrophysics Data System (ADS)

    Mokhtari, Marielle; Laurendeau, Denis

    1994-09-01

    A computer vision approach for the extraction of feature points on 3D images of dental imprints is presented. The position of feature points are needed for the measurement of a set of parameters for automatic diagnosis of malocclusion problems in orthodontics. The system for the acquisition of the 3D profile of the imprint, the procedure for the detection of the interstices between teeth, and the approach for the identification of the type of tooth are described, as well as the algorithm for the reconstruction of the surface of each type of tooth. A new approach for the detection of feature points, called the watershed algorithm, is described in detail. The algorithm is a two-stage procedure which tracks the position of local minima at four different scales and produces a final map of the position of the minima. Experimental results of the application of the watershed algorithm on actual 3D images of dental imprints are presented for molars, premolars and canines. The segmentation approach for the analysis of the shape of incisors is also described in detail.

  8. Light microscopy digital imaging.

    PubMed

    Joubert, James; Sharma, Deepak

    2011-10-01

    This unit presents an overview of digital imaging hardware used in light microscopy. CMOS, CCD, and EMCCDs are the primary sensors used. The strengths and weaknesses of each define the primary applications for these sensors. Sensor architecture and formats are also reviewed. Color camera design strategies and sensor window cleaning are also described in the unit.

  9. 3D Plant cell architecture of Arabidopsis thaliana (Brassicaceae) using focused ion beam–scanning electron microscopy1

    PubMed Central

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

    2014-01-01

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

  10. Performance prediction for 3D filtering of multichannel images

    NASA Astrophysics Data System (ADS)

    Rubel, Oleksii; Kozhemiakin, Ruslan A.; Abramov, Sergey K.; Lukin, Vladimir V.; Vozel, Benoit; Chehdi, Kacem

    2015-10-01

    Performance of denoising based on discrete cosine transform applied to multichannel remote sensing images corrupted by additive white Gaussian noise is analyzed. Images obtained by satellite Earth Observing-1 (EO-1) mission using hyperspectral imager instrument (Hyperion) that have high input SNR are taken as test images. Denoising performance is characterized by improvement of PSNR. For hard-thresholding 3D DCT-based denoising, simple statistics (probabilities to be less than a certain threshold) are used to predict denoising efficiency using curves fitted into scatterplots. It is shown that the obtained curves (approximations) provide prediction of denoising efficiency with high accuracy. Analysis is carried out for different numbers of channels processed jointly. Universality of prediction for different number of channels is proven.

  11. Phase Sensitive Cueing for 3D Objects in Overhead Images

    SciTech Connect

    Paglieroni, D W; Eppler, W G; Poland, D N

    2005-02-18

    A 3D solid model-aided object cueing method that matches phase angles of directional derivative vectors at image pixels to phase angles of vectors normal to projected model edges is described. It is intended for finding specific types of objects at arbitrary position and orientation in overhead images, independent of spatial resolution, obliqueness, acquisition conditions, and type of imaging sensor. It is shown that the phase similarity measure can be efficiently evaluated over all combinations of model position and orientation using the FFT. The highest degree of similarity over all model orientations is captured in a match surface of similarity values vs. model position. Unambiguous peaks in this surface are sorted in descending order of similarity value, and the small image thumbnails that contain them are presented to human analysts for inspection in sorted order.

  12. 3D Lunar Terrain Reconstruction from Apollo Images

    NASA Technical Reports Server (NTRS)

    Broxton, Michael J.; Nefian, Ara V.; Moratto, Zachary; Kim, Taemin; Lundy, Michael; Segal, Alkeksandr V.

    2009-01-01

    Generating accurate three dimensional planetary models is becoming increasingly important as NASA plans manned missions to return to the Moon in the next decade. This paper describes a 3D surface reconstruction system called the Ames Stereo Pipeline that is designed to produce such models automatically by processing orbital stereo imagery. We discuss two important core aspects of this system: (1) refinement of satellite station positions and pose estimates through least squares bundle adjustment; and (2) a stochastic plane fitting algorithm that generalizes the Lucas-Kanade method for optimal matching between stereo pair images.. These techniques allow us to automatically produce seamless, highly accurate digital elevation models from multiple stereo image pairs while significantly reducing the influence of image noise. Our technique is demonstrated on a set of 71 high resolution scanned images from the Apollo 15 mission

  13. 3D super-resolution imaging with blinking quantum dots.

    PubMed

    Wang, Yong; Fruhwirth, Gilbert; Cai, En; Ng, Tony; Selvin, Paul R

    2013-11-13

    Quantum dots are promising candidates for single molecule imaging due to their exceptional photophysical properties, including their intense brightness and resistance to photobleaching. They are also notorious for their blinking. Here we report a novel way to take advantage of quantum dot blinking to develop an imaging technique in three-dimensions with nanometric resolution. We first applied this method to simulated images of quantum dots and then to quantum dots immobilized on microspheres. We achieved imaging resolutions (fwhm) of 8-17 nm in the x-y plane and 58 nm (on coverslip) or 81 nm (deep in solution) in the z-direction, approximately 3-7 times better than what has been achieved previously with quantum dots. This approach was applied to resolve the 3D distribution of epidermal growth factor receptor (EGFR) molecules at, and inside of, the plasma membrane of resting basal breast cancer cells.

  14. Scattering robust 3D reconstruction via polarized transient imaging.

    PubMed

    Wu, Rihui; Suo, Jinli; Dai, Feng; Zhang, Yongdong; Dai, Qionghai

    2016-09-01

    Reconstructing 3D structure of scenes in the scattering medium is a challenging task with great research value. Existing techniques often impose strong assumptions on the scattering behaviors and are of limited performance. Recently, a low-cost transient imaging system has provided a feasible way to resolve the scene depth, by detecting the reflection instant on the time profile of a surface point. However, in cases with scattering medium, the rays are both reflected and scattered during transmission, and the depth calculated from the time profile largely deviates from the true value. To handle this problem, we used the different polarization behaviors of the reflection and scattering components, and introduced active polarization to separate the reflection component to estimate the scattering robust depth. Our experiments have demonstrated that our approach can accurately reconstruct the 3D structure underlying the scattering medium. PMID:27607944

  15. The 3D model control of image processing

    NASA Technical Reports Server (NTRS)

    Nguyen, An H.; Stark, Lawrence

    1989-01-01

    Telerobotics studies remote control of distant robots by a human operator using supervisory or direct control. Even if the robot manipulators has vision or other senses, problems arise involving control, communications, and delay. The communication delays that may be expected with telerobots working in space stations while being controlled from an Earth lab have led to a number of experiments attempting to circumvent the problem. This delay in communication is a main motivating factor in moving from well understood instantaneous hands-on manual control to less well understood supervisory control; the ultimate step would be the realization of a fully autonomous robot. The 3-D model control plays a crucial role in resolving many conflicting image processing problems that are inherent in resolving in the bottom-up approach of most current machine vision processes. The 3-D model control approach is also capable of providing the necessary visual feedback information for both the control algorithms and for the human operator.

  16. 3D Imaging of the OH mesospheric emissive layer

    NASA Astrophysics Data System (ADS)

    Kouahla, M. N.; Moreels, G.; Faivre, M.; Clairemidi, J.; Meriwether, J. W.; Lehmacher, G. A.; Vidal, E.; Veliz, O.

    2010-01-01

    A new and original stereo imaging method is introduced to measure the altitude of the OH nightglow layer and provide a 3D perspective map of the altitude of the layer centroid. Near-IR photographs of the OH layer are taken at two sites separated by a 645 km distance. Each photograph is processed in order to provide a satellite view of the layer. When superposed, the two views present a common diamond-shaped area. Pairs of matched points that correspond to a physical emissive point in the common area are identified in calculating a normalized cross-correlation coefficient (NCC). This method is suitable for obtaining 3D representations in the case of low-contrast objects. An observational campaign was conducted in July 2006 in Peru. The images were taken simultaneously at Cerro Cosmos (12°09‧08.2″ S, 75°33‧49.3″ W, altitude 4630 m) close to Huancayo and Cerro Verde Tellolo (16°33‧17.6″ S, 71°39‧59.4″ W, altitude 2272 m) close to Arequipa. 3D maps of the layer surface were retrieved and compared with pseudo-relief intensity maps of the same region. The mean altitude of the emission barycenter is located at 86.3 km on July 26. Comparable relief wavy features appear in the 3D and intensity maps. It is shown that the vertical amplitude of the wave system varies as exp (Δz/2H) within the altitude range Δz = 83.5-88.0 km, H being the scale height. The oscillatory kinetic energy at the altitude of the OH layer is comprised between 3 × 10-4 and 5.4 × 10-4 J/m3, which is 2-3 times smaller than the values derived from partial radio wave at 52°N latitude.

  17. 3D range scan enhancement using image-based methods

    NASA Astrophysics Data System (ADS)

    Herbort, Steffen; Gerken, Britta; Schugk, Daniel; Wöhler, Christian

    2013-10-01

    This paper addresses the problem of 3D surface scan refinement, which is desirable due to noise, outliers, and missing measurements being present in the 3D surfaces obtained with a laser scanner. We present a novel algorithm for the fusion of absolute laser scanner depth profiles and photometrically estimated surface normal data, which yields a noise-reduced and highly detailed depth profile with large scale shape robustness. In contrast to other approaches published in the literature, the presented algorithm (1) regards non-Lambertian surfaces, (2) simultaneously computes surface reflectance (i.e. BRDF) parameters required for 3D reconstruction, (3) models pixelwise incident light and viewing directions, and (4) accounts for interreflections. The algorithm as such relies on the minimization of a three-component error term, which penalizes intensity deviations, integrability deviations, and deviations from the known large-scale surface shape. The solution of the error minimization is obtained iteratively based on a calculus of variations. BRDF parameters are estimated by initially reducing and then iteratively refining the optical resolution, which provides the required robust data basis. The 3D reconstruction of concave surface regions affected by interreflections is improved by compensating global illumination in the image data. The algorithm is evaluated based on eight objects with varying albedos and reflectance behaviors (diffuse, specular, metallic). The qualitative evaluation shows a removal of outliers and a strong reduction of noise, while the large scale shape is preserved. Fine surface details Which are previously not contained in the surface scans, are incorporated through using image data. The algorithm is evaluated with respect to its absolute accuracy using two caliper objects of known shape, and based on synthetically generated data. The beneficial effect of interreflection compensation on the reconstruction accuracy is evaluated quantitatively in a

  18. Multiple 2D video/3D medical image registration algorithm

    NASA Astrophysics Data System (ADS)

    Clarkson, Matthew J.; Rueckert, Daniel; Hill, Derek L.; Hawkes, David J.

    2000-06-01

    In this paper we propose a novel method to register at least two vide images to a 3D surface model. The potential applications of such a registration method could be in image guided surgery, high precision radiotherapy, robotics or computer vision. Registration is performed by optimizing a similarity measure with respect to the pose parameters. The similarity measure is based on 'photo-consistency' and computes for each surface point, how consistent the corresponding video image information in each view is with a lighting model. We took four video views of a volunteer's face, and used an independent method to reconstruct a surface that was intrinsically registered to the four views. In addition, we extracted a skin surface from the volunteer's MR scan. The surfaces were misregistered from a gold standard pose and our algorithm was used to register both types of surfaces to the video images. For the reconstructed surface, the mean 3D error was 1.53 mm. For the MR surface, the standard deviation of the pose parameters after registration ranged from 0.12 to 0.70 mm and degrees. The performance of the algorithm is accurate, precise and robust.

  19. 3D seismic imaging on massively parallel computers

    SciTech Connect

    Womble, D.E.; Ober, C.C.; Oldfield, R.

    1997-02-01

    The ability to image complex geologies such as salt domes in the Gulf of Mexico and thrusts in mountainous regions is a key to reducing the risk and cost associated with oil and gas exploration. Imaging these structures, however, is computationally expensive. Datasets can be terabytes in size, and the processing time required for the multiple iterations needed to produce a velocity model can take months, even with the massively parallel computers available today. Some algorithms, such as 3D, finite-difference, prestack, depth migration remain beyond the capacity of production seismic processing. Massively parallel processors (MPPs) and algorithms research are the tools that will enable this project to provide new seismic processing capabilities to the oil and gas industry. The goals of this work are to (1) develop finite-difference algorithms for 3D, prestack, depth migration; (2) develop efficient computational approaches for seismic imaging and for processing terabyte datasets on massively parallel computers; and (3) develop a modular, portable, seismic imaging code.

  20. Improving 3D Wavelet-Based Compression of Hyperspectral Images

    NASA Technical Reports Server (NTRS)

    Klimesh, Matthew; Kiely, Aaron; Xie, Hua; Aranki, Nazeeh

    2009-01-01

    Two methods of increasing the effectiveness of three-dimensional (3D) wavelet-based compression of hyperspectral images have been developed. (As used here, images signifies both images and digital data representing images.) The methods are oriented toward reducing or eliminating detrimental effects of a phenomenon, referred to as spectral ringing, that is described below. In 3D wavelet-based compression, an image is represented by a multiresolution wavelet decomposition consisting of several subbands obtained by applying wavelet transforms in the two spatial dimensions corresponding to the two spatial coordinate axes of the image plane, and by applying wavelet transforms in the spectral dimension. Spectral ringing is named after the more familiar spatial ringing (spurious spatial oscillations) that can be seen parallel to and near edges in ordinary images reconstructed from compressed data. These ringing phenomena are attributable to effects of quantization. In hyperspectral data, the individual spectral bands play the role of edges, causing spurious oscillations to occur in the spectral dimension. In the absence of such corrective measures as the present two methods, spectral ringing can manifest itself as systematic biases in some reconstructed spectral bands and can reduce the effectiveness of compression of spatially-low-pass subbands. One of the two methods is denoted mean subtraction. The basic idea of this method is to subtract mean values from spatial planes of spatially low-pass subbands prior to encoding, because (a) such spatial planes often have mean values that are far from zero and (b) zero-mean data are better suited for compression by methods that are effective for subbands of two-dimensional (2D) images. In this method, after the 3D wavelet decomposition is performed, mean values are computed for and subtracted from each spatial plane of each spatially-low-pass subband. The resulting data are converted to sign-magnitude form and compressed in a

  1. Measuring surface topography with scanning electron microscopy. I. EZEImage: a program to obtain 3D surface data.

    PubMed

    Ponz, Ezequiel; Ladaga, Juan Luis; Bonetto, Rita Dominga

    2006-04-01

    Scanning electron microscopy (SEM) is widely used in the science of materials and different parameters were developed to characterize the surface roughness. In a previous work, we studied the surface topography with fractal dimension at low scale and two parameters at high scale by using the variogram, that is, variance vs. step log-log graph, of a SEM image. Those studies were carried out with the FERImage program, previously developed by us. To verify the previously accepted hypothesis by working with only an image, it is indispensable to have reliable three-dimensional (3D) surface data. In this work, a new program (EZEImage) to characterize 3D surface topography in SEM has been developed. It uses fast cross correlation and dynamic programming to obtain reliable dense height maps in a few seconds which can be displayed as an image where each gray level represents a height value. This image can be used for the FERImage program or any other software to obtain surface topography characteristics. EZEImage also generates anaglyph images as well as characterizes 3D surface topography by means of a parameter set to describe amplitude properties and three functional indices for characterizing bearing and fluid properties. PMID:17481354

  2. 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.

  3. Development of 3D microwave imaging reflectometry in LHD (invited).

    PubMed

    Nagayama, Y; Kuwahara, D; Yoshinaga, T; Hamada, Y; Kogi, Y; Mase, A; Tsuchiya, H; Tsuji-Iio, S; Yamaguchi, S

    2012-10-01

    Three-dimensional (3D) microwave imaging reflectometry has been developed in the large helical device to visualize fluctuating reflection surface which is caused by the density fluctuations. The plasma is illuminated by the probe wave with four frequencies, which correspond to four radial positions. The imaging optics makes the image of cut-off surface onto the 2D (7 × 7 channels) horn antenna mixer arrays. Multi-channel receivers have been also developed using micro-strip-line technology to handle many channels at reasonable cost. This system is first applied to observe the edge harmonic oscillation (EHO), which is an MHD mode with many harmonics that appears in the edge plasma. A narrow structure along field lines is observed during EHO.

  4. Density-tapered spiral arrays for ultrasound 3-D imaging.

    PubMed

    Ramalli, Alessandro; Boni, Enrico; Savoia, Alessandro Stuart; Tortoli, Piero

    2015-08-01

    The current high interest in 3-D ultrasound imaging is pushing the development of 2-D probes with a challenging number of active elements. The most popular approach to limit this number is the sparse array technique, which designs the array layout by means of complex optimization algorithms. These algorithms are typically constrained by a few steering conditions, and, as such, cannot guarantee uniform side-lobe performance at all angles. The performance may be improved by the ungridded extensions of the sparse array technique, but this result is achieved at the expense of a further complication of the optimization process. In this paper, a method to design the layout of large circular arrays with a limited number of elements according to Fermat's spiral seeds and spatial density modulation is proposed and shown to be suitable for application to 3-D ultrasound imaging. This deterministic, aperiodic, and balanced positioning procedure attempts to guarantee uniform performance over a wide range of steering angles. The capabilities of the method are demonstrated by simulating and comparing the performance of spiral and dense arrays. A good trade-off for small vessel imaging is found, e.g., in the 60λ spiral array with 1.0λ elements and Blackman density tapering window. Here, the grating lobe level is -16 dB, the lateral resolution is lower than 6λ the depth of field is 120λ and, the average contrast is 10.3 dB, while the sensitivity remains in a 5 dB range for a wide selection of steering angles. The simulation results may represent a reference guide to the design of spiral sparse array probes for different application fields. PMID:26285181

  5. 3D-LZ helicopter ladar imaging system

    NASA Astrophysics Data System (ADS)

    Savage, James; Harrington, Walter; McKinley, R. Andrew; Burns, H. N.; Braddom, Steven; Szoboszlay, Zoltan

    2010-04-01

    A joint-service team led by the Air Force Research Laboratory's Munitions and Sensors Directorates completed a successful flight test demonstration of the 3D-LZ Helicopter LADAR Imaging System. This was a milestone demonstration in the development of technology solutions for a problem known as "helicopter brownout", the loss of situational awareness caused by swirling sand during approach and landing. The 3D-LZ LADAR was developed by H.N. Burns Engineering and integrated with the US Army Aeroflightdynamics Directorate's Brown-Out Symbology System aircraft state symbology aboard a US Army EH-60 Black Hawk helicopter. The combination of these systems provided an integrated degraded visual environment landing solution with landing zone situational awareness as well as aircraft guidance and obstacle avoidance information. Pilots from the U.S. Army, Air Force, Navy, and Marine Corps achieved a 77% landing rate in full brownout conditions at a test range at Yuma Proving Ground, Arizona. This paper will focus on the LADAR technology used in 3D-LZ and the results of this milestone demonstration.

  6. Ultra-realistic 3-D imaging based on colour holography

    NASA Astrophysics Data System (ADS)

    Bjelkhagen, H. I.

    2013-02-01

    A review of recent progress in colour holography is provided with new applications. Colour holography recording techniques in silver-halide emulsions are discussed. Both analogue, mainly Denisyuk colour holograms, and digitally-printed colour holograms are described and their recent improvements. An alternative to silver-halide materials are the panchromatic photopolymer materials such as the DuPont and Bayer photopolymers which are covered. The light sources used to illuminate the recorded holograms are very important to obtain ultra-realistic 3-D images. In particular the new light sources based on RGB LEDs are described. They show improved image quality over today's commonly used halogen lights. Recent work in colour holography by holographers and companies in different countries around the world are included. To record and display ultra-realistic 3-D images with perfect colour rendering are highly dependent on the correct recording technique using the optimal recording laser wavelengths, the availability of improved panchromatic recording materials and combined with new display light sources.

  7. Image segmentation and 3D visualization for MRI mammography

    NASA Astrophysics Data System (ADS)

    Li, Lihua; Chu, Yong; Salem, Angela F.; Clark, Robert A.

    2002-05-01

    MRI mammography has a number of advantages, including the tomographic, and therefore three-dimensional (3-D) nature, of the images. It allows the application of MRI mammography to breasts with dense tissue, post operative scarring, and silicon implants. However, due to the vast quantity of images and subtlety of difference in MR sequence, there is a need for reliable computer diagnosis to reduce the radiologist's workload. The purpose of this work was to develop automatic breast/tissue segmentation and visualization algorithms to aid physicians in detecting and observing abnormalities in breast. Two segmentation algorithms were developed: one for breast segmentation, the other for glandular tissue segmentation. In breast segmentation, the MRI image is first segmented using an adaptive growing clustering method. Two tracing algorithms were then developed to refine the breast air and chest wall boundaries of breast. The glandular tissue segmentation was performed using an adaptive thresholding method, in which the threshold value was spatially adaptive using a sliding window. The 3D visualization of the segmented 2D slices of MRI mammography was implemented under IDL environment. The breast and glandular tissue rendering, slicing and animation were displayed.

  8. 3D Multispectral Light Propagation Model For Subcutaneous Veins Imaging

    SciTech Connect

    Paquit, Vincent C; Price, Jeffery R; Meriaudeau, Fabrice; Tobin Jr, Kenneth William

    2008-01-01

    In this paper, we describe a new 3D light propagation model aimed at understanding the effects of various physiological properties on subcutaneous vein imaging. In particular, we build upon the well known MCML (Monte Carlo Multi Layer) code and present a tissue model that improves upon the current state-of-the-art by: incorporating physiological variation, such as melanin concentration, fat content, and layer thickness; including veins of varying depth and diameter; using curved surfaces from real arm shapes; and modeling the vessel wall interface. We describe our model, present results from the Monte Carlo modeling, and compare these results with those obtained with other Monte Carlo methods.

  9. 3D laser optoacoustic ultrasonic imaging system for preclinical research

    NASA Astrophysics Data System (ADS)

    Ermilov, Sergey A.; Conjusteau, André; Hernandez, Travis; Su, Richard; Nadvoretskiy, Vyacheslav; Tsyboulski, Dmitri; Anis, Fatima; Anastasio, Mark A.; Oraevsky, Alexander A.

    2013-03-01

    In this work, we introduce a novel three-dimensional imaging system for in vivo high-resolution anatomical and functional whole-body visualization of small animal models developed for preclinical or other type of biomedical research. The system (LOUIS-3DM) combines a multi-wavelength optoacoustic and ultrawide-band laser ultrasound tomographies to obtain coregistered maps of tissue optical absorption and acoustic properties, displayed within the skin outline of the studied animal. The most promising applications of the LOUIS-3DM include 3D angiography, cancer research, and longitudinal studies of biological distribution of optoacoustic contrast agents (carbon nanotubes, metal plasmonic nanoparticles, etc.).

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

    PubMed Central

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

    2015-01-01

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

  12. 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

  13. Pinhole shifting lifetime imaging microscopy.

    PubMed

    Ramshesh, Venkat K; Lemasters, John J

    2008-01-01

    Lifetime imaging microscopy is a powerful tool to probe biological phenomena independent of luminescence intensity and fluorophore concentration. We describe time-resolved imaging of long-lifetime luminescence with an unmodified commercial laser scanning confocal/multiphoton microscope. The principle of the measurement is displacement of the detection pinhole to collect delayed luminescence from a position lagging the rasting laser beam. As proof of principle, luminescence from microspheres containing europium (Eu(3+)), a red emitting probe, was compared to that of short-lifetime green-fluorescing microspheres and/or fluorescein and rhodamine in solution. Using 720-nm two-photon excitation and a pinhole diameter of 1 Airy unit, the short-lifetime fluorescence of fluorescein, rhodamine and green microspheres disappeared much more rapidly than the long-lifetime phosphorescence of Eu(3+) microspheres as the pinhole was repositioned in the lagging direction. In contrast, repositioning of the pinhole in the leading and orthogonal directions caused equal loss of short- and long-lifetime luminescence. From measurements at different lag pinhole positions, a lifetime of 270 micros was estimated for the Eu(3+) microspheres, consistent with independent measurements. This simple adaptation is the basis for quantitative 3-D lifetime imaging microscopy. PMID:19123648

  14. Recent progress in 3-D imaging of sea freight containers

    SciTech Connect

    Fuchs, Theobald Schön, Tobias Sukowski, Frank; Dittmann, Jonas; Hanke, Randolf

    2015-03-31

    The inspection of very large objects like sea freight containers with X-ray Computed Tomography (CT) is an emerging technology. A complete 3-D CT scan of a see-freight container takes several hours. Of course, this is too slow to apply it to a large number of containers. However, the benefits of a 3-D CT for sealed freight are obvious: detection of potential threats or illicit cargo without being confronted with legal complications or high time consumption and risks for the security personnel during a manual inspection. Recently distinct progress was made in the field of reconstruction of projections with only a relatively low number of angular positions. Instead of today’s 500 to 1000 rotational steps, as needed for conventional CT reconstruction techniques, this new class of algorithms provides the potential to reduce the number of projection angles approximately by a factor of 10. The main drawback of these advanced iterative methods is the high consumption for numerical processing. But as computational power is getting steadily cheaper, there will be practical applications of these complex algorithms in a foreseeable future. In this paper, we discuss the properties of iterative image reconstruction algorithms and show results of their application to CT of extremely large objects scanning a sea-freight container. A specific test specimen is used to quantitatively evaluate the image quality in terms of spatial and contrast resolution and depending on different number of projections.

  15. Tactile-optical 3D sensor applying image processing

    NASA Astrophysics Data System (ADS)

    Neuschaefer-Rube, Ulrich; Wissmann, Mark

    2009-01-01

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

  16. Recent progress in 3-D imaging of sea freight containers

    NASA Astrophysics Data System (ADS)

    Fuchs, Theobald; Schön, Tobias; Dittmann, Jonas; Sukowski, Frank; Hanke, Randolf

    2015-03-01

    The inspection of very large objects like sea freight containers with X-ray Computed Tomography (CT) is an emerging technology. A complete 3-D CT scan of a see-freight container takes several hours. Of course, this is too slow to apply it to a large number of containers. However, the benefits of a 3-D CT for sealed freight are obvious: detection of potential threats or illicit cargo without being confronted with legal complications or high time consumption and risks for the security personnel during a manual inspection. Recently distinct progress was made in the field of reconstruction of projections with only a relatively low number of angular positions. Instead of today's 500 to 1000 rotational steps, as needed for conventional CT reconstruction techniques, this new class of algorithms provides the potential to reduce the number of projection angles approximately by a factor of 10. The main drawback of these advanced iterative methods is the high consumption for numerical processing. But as computational power is getting steadily cheaper, there will be practical applications of these complex algorithms in a foreseeable future. In this paper, we discuss the properties of iterative image reconstruction algorithms and show results of their application to CT of extremely large objects scanning a sea-freight container. A specific test specimen is used to quantitatively evaluate the image quality in terms of spatial and contrast resolution and depending on different number of projections.

  17. Quantitative validation of 3D image registration techniques

    NASA Astrophysics Data System (ADS)

    Holton Tainter, Kerrie S.; Taneja, Udita; Robb, Richard A.

    1995-05-01

    Multimodality images obtained from different medical imaging systems such as magnetic resonance (MR), computed tomography (CT), ultrasound (US), positron emission tomography (PET), single photon emission computed tomography (SPECT) provide largely complementary characteristic or diagnostic information. Therefore, it is an important research objective to `fuse' or combine this complementary data into a composite form which would provide synergistic information about the objects under examination. An important first step in the use of complementary fused images is 3D image registration, where multi-modality images are brought into spatial alignment so that the point-to-point correspondence between image data sets is known. Current research in the field of multimodality image registration has resulted in the development and implementation of several different registration algorithms, each with its own set of requirements and parameters. Our research has focused on the development of a general paradigm for measuring, evaluating and comparing the performance of different registration algorithms. Rather than evaluating the results of one algorithm under a specific set of conditions, we suggest a general approach to validation using simulation experiments, where the exact spatial relationship between data sets is known, along with phantom data, to characterize the behavior of an algorithm via a set of quantitative image measurements. This behavior may then be related to the algorithm's performance with real patient data, where the exact spatial relationship between multimodality images is unknown. Current results indicate that our approach is general enough to apply to several different registration algorithms. Our methods are useful for understanding the different sources of registration error and for comparing the results between different algorithms.

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

    PubMed

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

    2015-05-20

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

  19. Active and interactive floating image display using holographic 3D images

    NASA Astrophysics Data System (ADS)

    Morii, Tsutomu; Sakamoto, Kunio

    2006-08-01

    We developed a prototype tabletop holographic display system. This system consists of the object recognition system and the spatial imaging system. In this paper, we describe the recognition system using an RFID tag and the 3D display system using a holographic technology. A 3D display system is useful technology for virtual reality, mixed reality and augmented reality. We have researched spatial imaging and interaction system. We have ever proposed 3D displays using the slit as a parallax barrier, the lenticular screen and the holographic optical elements(HOEs) for displaying active image 1,2,3. The purpose of this paper is to propose the interactive system using these 3D imaging technologies. In this paper, the authors describe the interactive tabletop 3D display system. The observer can view virtual images when the user puts the special object on the display table. The key technologies of this system are the object recognition system and the spatial imaging display.

  20. High Resolution 3D Radar Imaging of Comet Interiors

    NASA Astrophysics Data System (ADS)

    Asphaug, E. I.; Gim, Y.; Belton, M.; Brophy, J.; Weissman, P. R.; Heggy, E.

    2012-12-01

    Knowing the interiors of comets and other primitive bodies is fundamental to our understanding of how planets formed. We have developed a Discovery-class mission formulation, Comet Radar Explorer (CORE), based on the use of previously flown planetary radar sounding techniques, with the goal of obtaining high resolution 3D images of the interior of a small primitive body. We focus on the Jupiter-Family Comets (JFCs) as these are among the most primitive bodies reachable by spacecraft. Scattered in from far beyond Neptune, they are ultimate targets of a cryogenic sample return mission according to the Decadal Survey. Other suitable targets include primitive NEOs, Main Belt Comets, and Jupiter Trojans. The approach is optimal for small icy bodies ~3-20 km diameter with spin periods faster than about 12 hours, since (a) navigation is relatively easy, (b) radar penetration is global for decameter wavelengths, and (c) repeated overlapping ground tracks are obtained. The science mission can be as short as ~1 month for a fast-rotating JFC. Bodies smaller than ~1 km can be globally imaged, but the navigation solutions are less accurate and the relative resolution is coarse. Larger comets are more interesting, but radar signal is unlikely to be reflected from depths greater than ~10 km. So, JFCs are excellent targets for a variety of reasons. We furthermore focus on the use of Solar Electric Propulsion (SEP) to rendezvous shortly after the comet's perihelion. This approach leaves us with ample power for science operations under dormant conditions beyond ~2-3 AU. This leads to a natural mission approach of distant observation, followed by closer inspection, terminated by a dedicated radar mapping orbit. Radar reflections are obtained from a polar orbit about the icy nucleus, which spins underneath. Echoes are obtained from a sounder operating at dual frequencies 5 and 15 MHz, with 1 and 10 MHz bandwidths respectively. The dense network of echoes is used to obtain global 3D

  1. Compensation of log-compressed images for 3-D ultrasound.

    PubMed

    Sanches, João M; Marques, Jorge S

    2003-02-01

    In this study, a Bayesian approach was used for 3-D reconstruction in the presence of multiplicative noise and nonlinear compression of the ultrasound (US) data. Ultrasound images are often considered as being corrupted by multiplicative noise (speckle). Several statistical models have been developed to represent the US data. However, commercial US equipment performs a nonlinear image compression that reduces the dynamic range of the US signal for visualization purposes. This operation changes the distribution of the image pixels, preventing a straightforward application of the models. In this paper, the nonlinear compression is explicitly modeled and considered in the reconstruction process, where the speckle noise present in the radio frequency (RF) US data is modeled with a Rayleigh distribution. The results obtained by considering the compression of the US data are then compared with those obtained assuming no compression. It is shown that the estimation performed using the nonlinear log-compression model leads to better results than those obtained with the Rayleigh reconstruction method. The proposed algorithm is tested with synthetic and real data and the results are discussed. The results have shown an improvement in the reconstruction results when the compression operation is included in the image formation model, leading to sharper images with enhanced anatomical details.

  2. Real-time cylindrical curvilinear 3-D ultrasound imaging.

    PubMed

    Pua, E C; Yen, J T; Smith, S W

    2003-07-01

    In patients who are obese or exhibit signs of pulmonary disease, standard transthoracic scanning may yield poor quality cardiac images. For these conditions, two-dimensional transesophageal echocardiography (TEE) is established as an essential diagnostic tool. Current techniques in transesophageal scanning, though, are limited by incomplete visualization of cardiac structures in close proximity to the transducer. Thus, we propose a 2D curvilinear array for 3D transesophageal echocardiography in order to widen the field of view and increase visualization close to the transducer face. In this project, a 440 channel 5 MHz two-dimensional array with a 12.6 mm aperture diameter on a flexible interconnect circuit has been molded to a 4 mm radius of curvature. A 75% element yield was achieved during fabrication and an average -6dB bandwidth of 30% was observed in pulse-echo tests. Using this transducer in conjunction with modifications to the beam former delay software and scan converter display software of the our 3D scanner, we obtained cylindrical real-time curvilinear volumetric scans of tissue phantoms, including a field of view of greater than 120 degrees in the curved, azimuth direction and 65 degrees phased array sector scans in the elevation direction. These images were achieved using a stepped subaperture across the cylindrical curvilinear direction of the transducer face and phased array sector scanning in the noncurved plane. In addition, real-time volume rendered images of a tissue mimicking phantom with holes ranging from 1 cm to less than 4 mm have been obtained. 3D color flow Doppler results have also been acquired. This configuration can theoretically achieve volumes displaying 180 degrees by 120 degrees. The transducer is also capable of obtaining images through a curvilinear stepped subaperture in azimuth in conjunction with a rectilinear stepped subaperture in elevation, further increasing the field of view close to the transducer face. Future work

  3. 3D Imaging of Nanoparticle Distribution in Biological Tissue by Laser-Induced Breakdown Spectroscopy

    NASA Astrophysics Data System (ADS)

    Gimenez, Y.; Busser, B.; Trichard, F.; Kulesza, A.; Laurent, J. M.; Zaun, V.; Lux, F.; Benoit, J. M.; Panczer, G.; Dugourd, P.; Tillement, O.; Pelascini, F.; Sancey, L.; Motto-Ros, V.

    2016-07-01

    Nanomaterials represent a rapidly expanding area of research with huge potential for future medical applications. Nanotechnology indeed promises to revolutionize diagnostics, drug delivery, gene therapy, and many other areas of research. For any biological investigation involving nanomaterials, it is crucial to study the behavior of such nano-objects within tissues to evaluate both their efficacy and their toxicity. Here, we provide the first account of 3D label-free nanoparticle imaging at the entire-organ scale. The technology used is known as laser-induced breakdown spectroscopy (LIBS) and possesses several advantages such as speed of operation, ease of use and full compatibility with optical microscopy. We then used two different but complementary approaches to achieve 3D elemental imaging with LIBS: a volume reconstruction of a sliced organ and in-depth analysis. This proof-of-concept study demonstrates the quantitative imaging of both endogenous and exogenous elements within entire organs and paves the way for innumerable applications.

  4. Graph-regularized 3D shape reconstruction from highly anisotropic and noisy images

    PubMed Central

    Heinrich, Stephanie; Drewe, Philipp; Lou, Xinghua; Umrania, Shefali; Rätsch, Gunnar

    2014-01-01

    Analysis of microscopy images can provide insight into many biological processes. One particularly challenging problem is cellular nuclear segmentation in highly anisotropic and noisy 3D image data. Manually localizing and segmenting each and every cellular nucleus is very time-consuming, which remains a bottleneck in large-scale biological experiments. In this work, we present a tool for automated segmentation of cellular nuclei from 3D fluorescent microscopic data. Our tool is based on state-of-the-art image processing and machine learning techniques and provides a user-friendly graphical user interface. We show that our tool is as accurate as manual annotation and greatly reduces the time for the registration. PMID:25866587

  5. An efficient way of high-contrast, quasi-3D cellular imaging: off-axis illumination.

    PubMed

    Hostounský, Zdenĕk; Pelc, Radek

    2006-07-31

    An imaging system enabling a convenient visualisation of cells and other small objects is presented. It represents an adaptation of the optical microscope condenser, accommodating a built-in edge (relief) diaphragm brought close to the condenser iris diaphragm and enabling high-contrast pseudo-relief (quasi-3D) imaging. The device broadens the family of available apparatus based on the off-axis (or anaxial, asymmetric, inclined, oblique, schlieren-type, sideband) illumination. The simplicity of the design makes the condenser a user-friendly, dedicated device delivering high-contrast quasi-3D images of phase objects. Those are nearly invisible under the ordinary (axial) illumination. The phase contrast microscopy commonly used in visualisation of phase objects does not deliver the quasi-3D effect and introduces a disturbing 'halo' effect around the edges. The performance of the device presented here is demonstrated on living cells and tissue replicas. High-contrast quasi-3D images of cell-free preparations of biological origin (paper fibres and microcrystals) are shown as well. PMID:16678908

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

    PubMed Central

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

    2015-01-01

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

  7. 3D texture analysis for classification of second harmonic generation images of human ovarian cancer

    PubMed Central

    Wen, Bruce; Campbell, Kirby R.; Tilbury, Karissa; Nadiarnykh, Oleg; Brewer, Molly A.; Patankar, Manish; Singh, Vikas; Eliceiri, Kevin. W.; Campagnola, Paul J.

    2016-01-01

    Remodeling of the collagen architecture in the extracellular matrix (ECM) has been implicated in ovarian cancer. To quantify these alterations we implemented a form of 3D texture analysis to delineate the fibrillar morphology observed in 3D Second Harmonic Generation (SHG) microscopy image data of normal (1) and high risk (2) ovarian stroma, benign ovarian tumors (3), low grade (4) and high grade (5) serous tumors, and endometrioid tumors (6). We developed a tailored set of 3D filters which extract textural features in the 3D image sets to build (or learn) statistical models of each tissue class. By applying k-nearest neighbor classification using these learned models, we achieved 83–91% accuracies for the six classes. The 3D method outperformed the analogous 2D classification on the same tissues, where we suggest this is due the increased information content. This classification based on ECM structural changes will complement conventional classification based on genetic profiles and can serve as an additional biomarker. Moreover, the texture analysis algorithm is quite general, as it does not rely on single morphological metrics such as fiber alignment, length, and width but their combined convolution with a customizable basis set. PMID:27767180

  8. An instrument for 3D x-ray nano-imaging

    SciTech Connect

    Holler, M.; Raabe, J.; Diaz, A.; Guizar-Sicairos, M.; Quitmann, C.; Menzel, A.; Bunk, O.

    2012-07-15

    We present an instrument dedicated to 3D scanning x-ray microscopy, allowing a sample to be precisely scanned through a beam while the angle of x-ray incidence can be changed. The position of the sample is controlled with respect to the beam-defining optics by laser interferometry. The instrument achieves a position stability better than 10 nm standard deviation. The instrument performance is assessed using scanning x-ray diffraction microscopy and we demonstrate a resolution of 18 nm in 2D imaging of a lithographic test pattern while the beam was defined by a pinhole of 3 {mu}m in diameter. In 3D on a test object of copper interconnects of a microprocessor, a resolution of 53 nm is achieved.

  9. Fast 3D subsurface imaging with stepped-frequency GPR

    NASA Astrophysics Data System (ADS)

    Masarik, Matthew P.; Burns, Joseph; Thelen, Brian T.; Sutter, Lena

    2015-05-01

    This paper investigates an algorithm for forming 3D images of the subsurface using stepped-frequency GPR data. The algorithm is specifically designed for a handheld GPR and therefore accounts for the irregular sampling pattern in the data and the spatially-variant air-ground interface by estimating an effective "ground-plane" and then registering the data to the plane. The algorithm efficiently solves the 4th-order polynomial for the Snell reflection points using a fully vectorized iterative scheme. The forward operator is implemented efficiently using an accelerated nonuniform FFT (Greengard and Lee, 2004); the adjoint operator is implemented efficiently using an interpolation step coupled with an upsampled FFT. The imaging is done as a linearized version of the full inverse problem, which is regularized using a sparsity constraint to reduce sidelobes and therefore improve image localization. Applying an appropriate sparsity constraint, the algorithm is able to eliminate most the surrounding clutter and sidelobes, while still rendering valuable image properties such as shape and size. The algorithm is applied to simulated data, controlled experimental data (made available by Dr. Waymond Scott, Georgia Institute of Technology), and government-provided data with irregular sampling and air-ground interface.

  10. 3D lung image retrieval using localized features

    NASA Astrophysics Data System (ADS)

    Depeursinge, Adrien; Zrimec, Tatjana; Busayarat, Sata; Müller, Henning

    2011-03-01

    The interpretation of high-resolution computed tomography (HRCT) images of the chest showing disorders of the lung tissue associated with interstitial lung diseases (ILDs) is time-consuming and requires experience. Whereas automatic detection and quantification of the lung tissue patterns showed promising results in several studies, its aid for the clinicians is limited to the challenge of image interpretation, letting the radiologists with the problem of the final histological diagnosis. Complementary to lung tissue categorization, providing visually similar cases using content-based image retrieval (CBIR) is in line with the clinical workflow of the radiologists. In a preliminary study, a Euclidean distance based on volume percentages of five lung tissue types was used as inter-case distance for CBIR. The latter showed the feasibility of retrieving similar histological diagnoses of ILD based on visual content, although no localization information was used for CBIR. However, to retrieve and show similar images with pathology appearing at a particular lung position was not possible. In this work, a 3D localization system based on lung anatomy is used to localize low-level features used for CBIR. When compared to our previous study, the introduction of localization features allows improving early precision for some histological diagnoses, especially when the region of appearance of lung tissue disorders is important.

  11. Research of Fast 3D Imaging Based on Multiple Mode

    NASA Astrophysics Data System (ADS)

    Chen, Shibing; Yan, Huimin; Ni, Xuxiang; Zhang, Xiuda; Wang, Yu

    2016-02-01

    Three-dimensional (3D) imaging has received increasingly extensive attention and has been widely used currently. Lots of efforts have been put on three-dimensional imaging method and system study, in order to meet fast and high accurate requirement. In this article, we realize a fast and high quality stereo matching algorithm on field programmable gate array (FPGA) using the combination of time-of-flight (TOF) camera and binocular camera. Images captured from the two cameras own a same spatial resolution, letting us use the depth maps taken by the TOF camera to figure initial disparity. Under the constraint of the depth map as the stereo pairs when comes to stereo matching, expected disparity of each pixel is limited within a narrow search range. In the meanwhile, using field programmable gate array (FPGA, altera cyclone IV series) concurrent computing we can configure multi core image matching system, thus doing stereo matching on embedded system. The simulation results demonstrate that it can speed up the process of stereo matching and increase matching reliability and stability, realize embedded calculation, expand application range.

  12. Brain surface maps from 3-D medical images

    NASA Astrophysics Data System (ADS)

    Lu, Jiuhuai; Hansen, Eric W.; Gazzaniga, Michael S.

    1991-06-01

    The anatomic and functional localization of brain lesions for neurologic diagnosis and brain surgery is facilitated by labeling the cortical surface in 3D images. This paper presents a method which extracts cortical contours from magnetic resonance (MR) image series and then produces a planar surface map which preserves important anatomic features. The resultant map may be used for manual anatomic localization as well as for further automatic labeling. Outer contours are determined on MR cross-sectional images by following the clear boundaries between gray matter and cerebral-spinal fluid, skipping over sulci. Carrying this contour below the surface by shrinking it along its normal produces an inner contour that alternately intercepts gray matter (sulci) and white matter along its length. This procedure is applied to every section in the set, and the image (grayscale) values along the inner contours are radially projected and interpolated onto a semi-cylindrical surface with axis normal to the slices and large enough to cover the whole brain. A planar map of the cortical surface results by flattening this cylindrical surface. The projection from inner contour to cylindrical surface is unique in the sense that different points on the inner contour correspond to different points on the cylindrical surface. As the outer contours are readily obtained by automatic segmentation, cortical maps can be made directly from an MR series.

  13. MIMO based 3D imaging system at 360 GHz

    NASA Astrophysics Data System (ADS)

    Herschel, R.; Nowok, S.; Zimmermann, R.; Lang, S. A.; Pohl, N.

    2016-05-01

    A MIMO radar imaging system at 360 GHz is presented as a part of the comprehensive approach of the European FP7 project TeraSCREEN, using multiple frequency bands for active and passive imaging. The MIMO system consists of 16 transmitter and 16 receiver antennas within one single array. Using a bandwidth of 30 GHz, a range resolution up to 5 mm is obtained. With the 16×16 MIMO system 256 different azimuth bins can be distinguished. Mechanical beam steering is used to measure 130 different elevation angles where the angular resolution is obtained by a focusing elliptical mirror. With this system a high resolution 3D image can be generated with 4 frames per second, each containing 16 million points. The principle of the system is presented starting from the functional structure, covering the hardware design and including the digital image generation. This is supported by simulated data and discussed using experimental results from a preliminary 90 GHz system underlining the feasibility of the approach.

  14. Fast 3-D Tomographic Microwave Imaging for Breast Cancer Detection

    PubMed Central

    Meaney, Paul M.; Kaufman, Peter A.; diFlorio-Alexander, Roberta M.; Paulsen, Keith D.

    2013-01-01

    Microwave breast imaging (using electromagnetic waves of frequencies around 1 GHz) has mostly remained at the research level for the past decade, gaining little clinical acceptance. The major hurdles limiting patient use are both at the hardware level (challenges in collecting accurate and noncorrupted data) and software level (often plagued by unrealistic reconstruction times in the tens of hours). In this paper we report improvements that address both issues. First, the hardware is able to measure signals down to levels compatible with sub-centimeter image resolution while keeping an exam time under 2 min. Second, the software overcomes the enormous time burden and produces similarly accurate images in less than 20 min. The combination of the new hardware and software allows us to produce and report here the first clinical 3-D microwave tomographic images of the breast. Two clinical examples are selected out of 400+ exams conducted at the Dartmouth Hitchcock Medical Center (Lebanon, NH). The first example demonstrates the potential usefulness of our system for breast cancer screening while the second example focuses on therapy monitoring. PMID:22562726

  15. 3D Chemical and Elemental Imaging by STXM Spectrotomography

    NASA Astrophysics Data System (ADS)

    Wang, J.; Hitchcock, A. P.; Karunakaran, C.; Prange, A.; Franz, B.; Harkness, T.; Lu, Y.; Obst, M.; Hormes, J.

    2011-09-01

    Spectrotomography based on the scanning transmission x-ray microscope (STXM) at the 10ID-1 spectromicroscopy beamline of the Canadian Light Source was used to study two selected unicellular microorganisms. Spatial distributions of sulphur globules, calcium, protein, and polysaccharide in sulphur-metabolizing bacteria (Allochromatium vinosum) were determined at the S 2p, C 1s, and Ca 2p edges. 3D chemical mapping showed that the sulphur globules are located inside the bacteria with a strong spatial correlation with calcium ions (it is most probably calcium carbonate from the medium; however, with STXM the distribution and localization in the cell can be made visible, which is very interesting for a biologist) and polysaccharide-rich polymers, suggesting an influence of the organic components on the formation of the sulphur and calcium deposits. A second study investigated copper accumulating in yeast cells (Saccharomyces cerevisiae) treated with copper sulphate. 3D elemental imaging at the Cu 2p edge showed that Cu(II) is reduced to Cu(I) on the yeast cell wall. A novel needle-like wet cell sample holder for STXM spectrotomography studies of fully hydrated samples is discussed.

  16. 3D Chemical and Elemental Imaging by STXM Spectrotomography

    SciTech Connect

    Wang, J.; Karunakaran, C.; Lu, Y.; Hormes, J.; Hitchcock, A. P.; Prange, A.; Franz, B.; Harkness, T.; Obst, M.

    2011-09-09

    Spectrotomography based on the scanning transmission x-ray microscope (STXM) at the 10ID-1 spectromicroscopy beamline of the Canadian Light Source was used to study two selected unicellular microorganisms. Spatial distributions of sulphur globules, calcium, protein, and polysaccharide in sulphur-metabolizing bacteria (Allochromatium vinosum) were determined at the S 2p, C 1s, and Ca 2p edges. 3D chemical mapping showed that the sulphur globules are located inside the bacteria with a strong spatial correlation with calcium ions (it is most probably calcium carbonate from the medium; however, with STXM the distribution and localization in the cell can be made visible, which is very interesting for a biologist) and polysaccharide-rich polymers, suggesting an influence of the organic components on the formation of the sulphur and calcium deposits. A second study investigated copper accumulating in yeast cells (Saccharomyces cerevisiae) treated with copper sulphate. 3D elemental imaging at the Cu 2p edge showed that Cu(II) is reduced to Cu(I) on the yeast cell wall. A novel needle-like wet cell sample holder for STXM spectrotomography studies of fully hydrated samples is discussed.

  17. An Efficient 3D Imaging using Structured Light Systems

    NASA Astrophysics Data System (ADS)

    Lee, Deokwoo

    Structured light 3D surface imaging has been crucial in the fields of image processing and computer vision, particularly in reconstruction, recognition and others. In this dissertation, we propose the approaches to development of an efficient 3D surface imaging system using structured light patterns including reconstruction, recognition and sampling criterion. To achieve an efficient reconstruction system, we address the problem in its many dimensions. In the first, we extract geometric 3D coordinates of an object which is illuminated by a set of concentric circular patterns and reflected to a 2D image plane. The relationship between the original and the deformed shape of the light patterns due to a surface shape provides sufficient 3D coordinates information. In the second, we consider system efficiency. The efficiency, which can be quantified by the size of data, is improved by reducing the number of circular patterns to be projected onto an object of interest. Akin to the Shannon-Nyquist Sampling Theorem, we derive the minimum number of circular patterns which sufficiently represents the target object with no considerable information loss. Specific geometric information (e.g. the highest curvature) of an object is key to deriving the minimum sampling density. In the third, the object, represented using the minimum number of patterns, has incomplete color information (i.e. color information is given a priori along with the curves). An interpolation is carried out to complete the photometric reconstruction. The results can be approximately reconstructed because the minimum number of the patterns may not exactly reconstruct the original object. But the result does not show considerable information loss, and the performance of an approximate reconstruction is evaluated by performing recognition or classification. In an object recognition, we use facial curves which are deformed circular curves (patterns) on a target object. We simply carry out comparison between the

  18. 3D imaging of semiconductor components by discrete laminography

    SciTech Connect

    Batenburg, K. J.; Palenstijn, W. J.; Sijbers, J.

    2014-06-19

    X-ray laminography is a powerful technique for quality control of semiconductor components. Despite the advantages of nondestructive 3D imaging over 2D techniques based on sectioning, the acquisition time is still a major obstacle for practical use of the technique. In this paper, we consider the application of Discrete Tomography to laminography data, which can potentially reduce the scanning time while still maintaining a high reconstruction quality. By incorporating prior knowledge in the reconstruction algorithm about the materials present in the scanned object, far more accurate reconstructions can be obtained from the same measured data compared to classical reconstruction methods. We present a series of simulation experiments that illustrate the potential of the approach.

  19. Near field 3D scene simulation for passive microwave imaging

    NASA Astrophysics Data System (ADS)

    Zhang, Cheng; Wu, Ji

    2006-10-01

    Scene simulation is a necessary work in near field passive microwave remote sensing. A 3-D scene simulation model of microwave radiometric imaging based on ray tracing method is present in this paper. The essential influencing factors and general requirements are considered in this model such as the rough surface radiation, the sky radiation witch act as the uppermost illuminator in out door circumstance, the polarization rotation of the temperature rays caused by multiple reflections, and the antenna point spread function witch determines the resolution of the model final outputs. Using this model we simulate a virtual scene and analyzed the appeared microwave radiometric phenomenology, at last two real scenes of building and airstrip were simulated for validating the model. The comparison between the simulation and field measurements indicates that this model is completely feasible in practice. Furthermore, we analyzed the signatures of model outputs, and achieved some underlying phenomenology of microwave radiation witch is deferent with that in optical and infrared bands.

  20. Needle placement for piriformis injection using 3-D imaging.

    PubMed

    Clendenen, Steven R; Candler, Shawn A; Osborne, Michael D; Palmer, Scott C; Duench, Stephanie; Glynn, Laura; Ghazi, Salim M

    2013-01-01

    Piriformis syndrome is a pain syndrome originating in the buttock and is attributed to 6% - 8% of patients referred for the treatment of back and leg pain. The treatment for piriformis syndrome using fluoroscopy, computed tomography (CT), electromyography (EMG), and ultrasound (US) has become standard practice. The treatment of Piriformis Syndrome has evolved to include fluoroscopy and EMG with CT guidance. We present a case study of 5 successful piriformis injections using 3-D computer-assisted electromagnet needle tracking coupled with ultrasound. A 6-degree of freedom electromagnetic position tracker was attached to the ultrasound probe that allowed the system to detect the position and orientation of the probe in the magnetic field. The tracked ultrasound probe was used to find the posterior superior iliac spine. Subsequently, 3 points were captured to register the ultrasound image with the CT or magnetic resonance image scan. Moreover, after the registration was obtained, the navigation system visualized the tracked needle relative to the CT scan in real-time using 2 orthogonal multi-planar reconstructions centered at the tracked needle tip. Conversely, a recent study revealed that fluoroscopically guided injections had 30% accuracy compared to ultrasound guided injections, which tripled the accuracy percentage. This novel technique exhibited an accurate needle guidance injection precision of 98% while advancing to the piriformis muscle and avoiding the sciatic nerve. The mean (± SD) procedure time was 19.08 (± 4.9) minutes. This technique allows for electromagnetic instrument tip tracking with real-time 3-D guidance to the selected target. As with any new technique, a learning curve is expected; however, this technique could offer an alternative, minimizing radiation exposure. PMID:23703429

  1. NMR imaging microscopy

    SciTech Connect

    Not Available

    1986-10-01

    In the past several years, proton nuclear magnetic resonance (NMR) imaging has become an established technique in diagnostic medicine and biomedical research. Although much of the work in this field has been directed toward development of whole-body imagers, James Aguayo, Stephen Blackband, and Joseph Schoeninger of the Johns Hopkins University School of Medicine working with Markus Hintermann and Mark Mattingly of Bruker Medical Instruments, recently developed a small-bore NMR microscope with sufficient resolution to image a single African clawed toad cell (Nature 1986, 322, 190-91). This improved resolution should lead to increased use of NMR imaging for chemical, as well as biological or physiological, applications. The future of NMR microscopy, like that of many other newly emerging techniques, is ripe with possibilities. Because of its high cost, however, it is likely to remain primarily a research tool for some time. ''It's like having a camera,'' says Smith. ''You've got a way to look at things at very fine levels, and people are going to find lots of uses for it. But it is a very expensive technique - it costs $100,000 to add imaging capability once you have a high-resolution NMR, which itself is at least a $300,000 instrument. If it can answer even a few questions that can't be answered any other way, though, it may be well worth the cost.''

  2. 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

  3. Multi Length Scale Imaging of Flocculated Estuarine Sediments; Insights into their Complex 3D Structure

    NASA Astrophysics Data System (ADS)

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

    2015-04-01

    Suspended estuarine sediments form flocs that are compositionally complex, fragile and irregularly shaped. The fate and transport of suspended particulate matter (SPM) is determined by the size, shape, density, porosity and stability of these flocs and prediction of SPM transport requires accurate measurements of these three-dimensional (3D) physical properties. However, the multi-scaled nature of flocs in addition to their fragility makes their characterisation in 3D problematic. Correlative microscopy is a strategy involving the spatial registration of information collected at different scales using several imaging modalities. Previously, conventional optical microscopy (COM) and transmission electron microscopy (TEM) have enabled 2-dimensional (2D) floc characterisation at the gross (> 1 µm) and sub-micron scales respectively. Whilst this has proven insightful there remains a critical spatial and dimensional gap preventing the accurate measurement of geometric properties and an understanding of how structures at different scales are related. Within life sciences volumetric imaging techniques such as 3D micro-computed tomography (3D µCT) and focused ion beam scanning electron microscopy [FIB-SEM (or FIB-tomography)] have been combined to characterise materials at the centimetre to micron scale. Combining these techniques with TEM enables an advanced correlative study, allowing material properties across multiple spatial and dimensional scales to be visualised. The aims of this study are; 1) to formulate an advanced correlative imaging strategy combining 3D µCT, FIB-tomography and TEM; 2) to acquire 3D datasets; 3) to produce a model allowing their co-visualisation; 4) to interpret 3D floc structure. To reduce the chance of structural alterations during analysis samples were first 'fixed' in 2.5% glutaraldehyde/2% formaldehyde before being embedding in Durcupan resin. Intermediate steps were implemented to improve contrast and remove pore water, achieved by the

  4. Machine Learning of Hierarchical Clustering to Segment 2D and 3D Images

    PubMed Central

    Nunez-Iglesias, Juan; Kennedy, Ryan; Parag, Toufiq; Shi, Jianbo; Chklovskii, Dmitri B.

    2013-01-01

    We aim to improve segmentation through the use of machine learning tools during region agglomeration. We propose an active learning approach for performing hierarchical agglomerative segmentation from superpixels. Our method combines multiple features at all scales of the agglomerative process, works for data with an arbitrary number of dimensions, and scales to very large datasets. We advocate the use of variation of information to measure segmentation accuracy, particularly in 3D electron microscopy (EM) images of neural tissue, and using this metric demonstrate an improvement over competing algorithms in EM and natural images. PMID:23977123

  5. GPU-accelerated denoising of 3D magnetic resonance images

    SciTech Connect

    Howison, Mark; Wes Bethel, E.

    2014-05-29

    The raw computational power of GPU accelerators enables fast denoising of 3D MR images using bilateral filtering, anisotropic diffusion, and non-local means. In practice, applying these filtering operations requires setting multiple parameters. This study was designed to provide better guidance to practitioners for choosing the most appropriate parameters by answering two questions: what parameters yield the best denoising results in practice? And what tuning is necessary to achieve optimal performance on a modern GPU? To answer the first question, we use two different metrics, mean squared error (MSE) and mean structural similarity (MSSIM), to compare denoising quality against a reference image. Surprisingly, the best improvement in structural similarity with the bilateral filter is achieved with a small stencil size that lies within the range of real-time execution on an NVIDIA Tesla M2050 GPU. Moreover, inappropriate choices for parameters, especially scaling parameters, can yield very poor denoising performance. To answer the second question, we perform an autotuning study to empirically determine optimal memory tiling on the GPU. The variation in these results suggests that such tuning is an essential step in achieving real-time performance. These results have important implications for the real-time application of denoising to MR images in clinical settings that require fast turn-around times.

  6. Spectral ladar: towards active 3D multispectral imaging

    NASA Astrophysics Data System (ADS)

    Powers, Michael A.; Davis, Christopher C.

    2010-04-01

    In this paper we present our Spectral LADAR concept, an augmented implementation of traditional LADAR. This sensor uses a polychromatic source to obtain range-resolved 3D spectral images which are used to identify objects based on combined spatial and spectral features, resolving positions in three dimensions and up to hundreds of meters in distance. We report on a proof-of-concept Spectral LADAR demonstrator that generates spectral point clouds from static scenes. The demonstrator transmits nanosecond supercontinuum pulses generated in a photonic crystal fiber. Currently we use a rapidly tuned receiver with a high-speed InGaAs APD for 25 spectral bands with the future expectation of implementing a linear APD array spectrograph. Each spectral band is independently range resolved with multiple return pulse recognition. This is a critical feature, enabling simultaneous spectral and spatial unmixing of partially obscured objects when not achievable using image fusion of monochromatic LADAR and passive spectral imagers. This enables higher identification confidence in highly cluttered environments such as forested or urban areas (e.g. vehicles behind camouflage or foliage). These environments present challenges for situational awareness and robotic perception which can benefit from the unique attributes of Spectral LADAR. Results from this demonstrator unit are presented for scenes typical of military operations and characterize the operation of the device. The results are discussed here in the context of autonomous vehicle navigation and target recognition.

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

    PubMed Central

    Tadrous, Paul Joseph

    2012-01-01

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

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

    DOE PAGES

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

    2016-05-01

    Here, we have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single moleculemore » super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.« less

  9. 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.

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

    PubMed Central

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

    2016-01-01

    We have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet. PMID:27375939

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

    SciTech Connect

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

    2016-01-01

    Here, we have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.

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

    PubMed

    Meddens, Marjolein B M; Liu, Sheng; Finnegan, Patrick S; Edwards, Thayne L; James, Conrad D; Lidke, Keith A

    2016-06-01

    We have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet. PMID:27375939

  13. Performance assessment of 3D surface imaging technique for medical imaging applications

    NASA Astrophysics Data System (ADS)

    Li, Tuotuo; Geng, Jason; Li, Shidong

    2013-03-01

    Recent development in optical 3D surface imaging technologies provide better ways to digitalize the 3D surface and its motion in real-time. The non-invasive 3D surface imaging approach has great potential for many medical imaging applications, such as motion monitoring of radiotherapy, pre/post evaluation of plastic surgery and dermatology, to name a few. Various commercial 3D surface imaging systems have appeared on the market with different dimension, speed and accuracy. For clinical applications, the accuracy, reproducibility and robustness across the widely heterogeneous skin color, tone, texture, shape properties, and ambient lighting is very crucial. Till now, a systematic approach for evaluating the performance of different 3D surface imaging systems still yet exist. In this paper, we present a systematic performance assessment approach to 3D surface imaging system assessment for medical applications. We use this assessment approach to exam a new real-time surface imaging system we developed, dubbed "Neo3D Camera", for image-guided radiotherapy (IGRT). The assessments include accuracy, field of view, coverage, repeatability, speed and sensitivity to environment, texture and color.

  14. 3D Slicer as an Image Computing Platform for the Quantitative Imaging Network

    PubMed Central

    Fedorov, Andriy; Beichel, Reinhard; Kalpathy-Cramer, Jayashree; Finet, Julien; Fillion-Robin, Jean-Christophe; Pujol, Sonia; Bauer, Christian; Jennings, Dominique; Fennessy, Fiona; Sonka, Milan; Buatti, John; Aylward, Stephen; Miller, James V.; Pieper, Steve; Kikinis, Ron

    2012-01-01

    Quantitative analysis has tremendous but mostly unrealized potential in healthcare to support objective and accurate interpretation of the clinical imaging. In 2008, the National Cancer Institute began building the Quantitative Imaging Network (QIN) initiative with the goal of advancing quantitative imaging in the context of personalized therapy and evaluation of treatment response. Computerized analysis is an important component contributing to reproducibility and efficiency of the quantitative imaging techniques. The success of quantitative imaging is contingent on robust analysis methods and software tools to bring these methods from bench to bedside. 3D Slicer is a free open source software application for medical image computing. As a clinical research tool, 3D Slicer is similar to a radiology workstation that supports versatile visualizations but also provides advanced functionality such as automated segmentation and registration for a variety of application domains. Unlike a typical radiology workstation, 3D Slicer is free and is not tied to specific hardware. As a programming platform, 3D Slicer facilitates translation and evaluation of the new quantitative methods by allowing the biomedical researcher to focus on the implementation of the algorithm, and providing abstractions for the common tasks of data communication, visualization and user interface development. Compared to other tools that provide aspects of this functionality, 3D Slicer is fully open source and can be readily extended and redistributed. In addition, 3D Slicer is designed to facilitate the development of new functionality in the form of 3D Slicer extensions. In this paper, we present an overview of 3D Slicer as a platform for prototyping, development and evaluation of image analysis tools for clinical research applications. To illustrate the utility of the platform in the scope of QIN, we discuss several use cases of 3D Slicer by the existing QIN teams, and we elaborate on the future

  15. 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.

  16. 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

  17. ROIC for gated 3D imaging LADAR receiver

    NASA Astrophysics Data System (ADS)

    Chen, Guoqiang; Zhang, Junling; Wang, Pan; Zhou, Jie; Gao, Lei; Ding, Ruijun

    2013-09-01

    Time of flight laser range finding, deep space communications and scanning video imaging are three applications requiring very low noise optical receivers to achieve detection of fast and weak optical signal. HgCdTe electrons initiated avalanche photodiodes (e-APDs) in linear multiplication mode is the detector of choice thanks to its high quantum efficiency, high gain at low bias, high bandwidth and low noise factor. In this project, a readout integrated circuit of hybrid e-APD focal plane array (FPA) with 100um pitch for 3D-LADAR was designed for gated optical receiver. The ROIC works at 77K, including unit cell circuit, column-level circuit, timing control, bias circuit and output driver. The unit cell circuit is a key component, which consists of preamplifier, correlated double Sampling (CDS), bias circuit and timing control module. Specially, the preamplifier used the capacitor feedback transimpedance amplifier (CTIA) structure which has two capacitors to offer switchable capacitance for passive/active dual mode imaging. The main circuit of column-level circuit is a precision Multiply-by-Two circuit which is implemented by switched-capacitor circuit. Switched-capacitor circuit is quite suitable for the signal processing of readout integrated circuit (ROIC) due to the working characteristics. The output driver uses a simply unity-gain buffer. Because the signal is amplified in column-level circuit, the amplifier in unity-gain buffer uses a rail-rail amplifier. In active imaging mode, the integration time is 80ns. Integrating current from 200nA to 4uA, this circuit shows the nonlinearity is less than 1%. In passive imaging mode, the integration time is 150ns. Integrating current from 1nA to 20nA shows the nonlinearity less than 1%.

  18. Deformable M-Reps for 3D Medical Image Segmentation.

    PubMed

    Pizer, Stephen M; Fletcher, P Thomas; Joshi, Sarang; Thall, Andrew; Chen, James Z; Fridman, Yonatan; Fritsch, Daniel S; Gash, Graham; Glotzer, John M; Jiroutek, Michael R; Lu, Conglin; Muller, Keith E; Tracton, Gregg; Yushkevich, Paul; Chaney, Edward L

    2003-11-01

    M-reps (formerly called DSLs) are a multiscale medial means for modeling and rendering 3D solid geometry. They are particularly well suited to model anatomic objects and in particular to capture prior geometric information effectively in deformable models segmentation approaches. The representation is based on figural models, which define objects at coarse scale by a hierarchy of figures - each figure generally a slab representing a solid region and its boundary simultaneously. This paper focuses on the use of single figure models to segment objects of relatively simple structure. A single figure is a sheet of medial atoms, which is interpolated from the model formed by a net, i.e., a mesh or chain, of medial atoms (hence the name m-reps), each atom modeling a solid region via not only a position and a width but also a local figural frame giving figural directions and an object angle between opposing, corresponding positions on the boundary implied by the m-rep. The special capability of an m-rep is to provide spatial and orientational correspondence between an object in two different states of deformation. This ability is central to effective measurement of both geometric typicality and geometry to image match, the two terms of the objective function optimized in segmentation by deformable models. The other ability of m-reps central to effective segmentation is their ability to support segmentation at multiple levels of scale, with successively finer precision. Objects modeled by single figures are segmented first by a similarity transform augmented by object elongation, then by adjustment of each medial atom, and finally by displacing a dense sampling of the m-rep implied boundary. While these models and approaches also exist in 2D, we focus on 3D objects. The segmentation of the kidney from CT and the hippocampus from MRI serve as the major examples in this paper. The accuracy of segmentation as compared to manual, slice-by-slice segmentation is reported.

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

    PubMed

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

    2016-02-01

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

  20. 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.

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

    PubMed

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

    2016-02-01

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

  2. 3D imaging of enzymes working in situ.

    PubMed

    Jamme, F; Bourquin, D; Tawil, G; Viksø-Nielsen, A; Buléon, A; Réfrégiers, M

    2014-06-01

    Today, development of slowly digestible food with positive health impact and production of biofuels is a matter of intense research. The latter is achieved via enzymatic hydrolysis of starch or biomass such as lignocellulose. Free label imaging, using UV autofluorescence, provides a great tool to follow one single enzyme when acting on a non-UV-fluorescent substrate. In this article, we report synchrotron DUV fluorescence in 3-dimensional imaging to visualize in situ the diffusion of enzymes on solid substrate. The degradation pathway of single starch granules by two amylases optimized for biofuel production and industrial starch hydrolysis was followed by tryptophan autofluorescence (excitation at 280 nm, emission filter at 350 nm). The new setup has been specially designed and developed for a 3D representation of the enzyme-substrate interaction during hydrolysis. Thus, this tool is particularly effective for improving knowledge and understanding of enzymatic hydrolysis of solid substrates such as starch and lignocellulosic biomass. It could open up the way to new routes in the field of green chemistry and sustainable development, that is, in biotechnology, biorefining, or biofuels. PMID:24796213

  3. Complex adaptation-based LDR image rendering for 3D image reconstruction

    NASA Astrophysics Data System (ADS)

    Lee, Sung-Hak; Kwon, Hyuk-Ju; Sohng, Kyu-Ik

    2014-07-01

    A low-dynamic tone-compression technique is developed for realistic image rendering that can make three-dimensional (3D) images similar to realistic scenes by overcoming brightness dimming in the 3D display mode. The 3D surround provides varying conditions for image quality, illuminant adaptation, contrast, gamma, color, sharpness, and so on. In general, gain/offset adjustment, gamma compensation, and histogram equalization have performed well in contrast compression; however, as a result of signal saturation and clipping effects, image details are removed and information is lost on bright and dark areas. Thus, an enhanced image mapping technique is proposed based on space-varying image compression. The performance of contrast compression is enhanced with complex adaptation in a 3D viewing surround combining global and local adaptation. Evaluating local image rendering in view of tone and color expression, noise reduction, and edge compensation confirms that the proposed 3D image-mapping model can compensate for the loss of image quality in the 3D mode.

  4. Site-specific 3D imaging of cells and tissues with a dual beam microscope

    PubMed Central

    Heymann, Jurgen A.W.; Hayles, Mike; Gestmann, Ingo; Giannuzzi, Lucille A.; Lich, Ben; Subramaniam, Sriram

    2006-01-01

    Current approaches to 3D imaging at subcellular resolution using confocal microscopy and electron tomography, while powerful, are limited to relatively thin and transparent specimens. Here we report on the use of a new generation of dual beam electron microscopes capable of site-specific imaging of the interior of cellular and tissue specimens at spatial resolutions about an order of magnitude better than those currently achieved with optical microscopy. The principle of imaging is based on using a focused ion beam to create a cut at a designated site in the specimen, followed by viewing the newly generated surface with a scanning electron beam. Iteration of these two steps several times thus results in the generation of a series of surface maps of the specimen at regularly spaced intervals, which can be converted into a three-dimensional map of the specimen. We have explored the potential of this sequential “slice-and-view” strategy for site-specific 3D imaging of frozen yeast cells and tumor tissue, and establish that this approach can identify the locations of intracellular features such as the 100 nm-wide yeast nuclear pore complex. We also show that 200 nm thick sections can be generated in situ by “milling” of resin-embedded specimens using the ion beam, providing a valuable alternative to manual sectioning of cells and tissues using an ultramicrotome. Our results demonstrate that dual beam imaging is a powerful new tool for cellular and subcellular imaging in 3D for both basic biomedical and clinical applications. PMID:16713294

  5. Analysis and dynamic 3D visualization of cerebral blood flow combining 3D and 4D MR image sequences

    NASA Astrophysics Data System (ADS)

    Forkert, Nils Daniel; Säring, Dennis; Fiehler, Jens; Illies, Till; Möller, Dietmar; Handels, Heinz

    2009-02-01

    In this paper we present a method for the dynamic visualization of cerebral blood flow. Spatio-temporal 4D magnetic resonance angiography (MRA) image datasets and 3D MRA datasets with high spatial resolution were acquired for the analysis of arteriovenous malformations (AVMs). One of the main tasks is the combination of the information of the 3D and 4D MRA image sequences. Initially, in the 3D MRA dataset the vessel system is segmented and a 3D surface model is generated. Then, temporal intensity curves are analyzed voxelwise in the 4D MRA image sequences. A curve fitting of the temporal intensity curves to a patient individual reference curve is used to extract the bolus arrival times in the 4D MRA sequences. After non-linear registration of both MRA datasets the extracted hemodynamic information is transferred to the surface model where the time points of inflow can be visualized color coded dynamically over time. The dynamic visualizations computed using the curve fitting method for the estimation of the bolus arrival times were rated superior compared to those computed using conventional approaches for bolus arrival time estimation. In summary the procedure suggested allows a dynamic visualization of the individual hemodynamic situation and better understanding during the visual evaluation of cerebral vascular diseases.

  6. Improvements of 3-D image quality in integral display by reducing distortion errors

    NASA Astrophysics Data System (ADS)

    Kawakita, Masahiro; Sasaki, Hisayuki; Arai, Jun; Okano, Fumio; Suehiro, Koya; Haino, Yasuyuki; Yoshimura, Makoto; Sato, Masahito

    2008-02-01

    An integral three-dimensional (3-D) system based on the principle of integral photography can display natural 3-D images. We studied ways of improving the resolution and viewing angle of 3-D images by using extremely highresolution (EHR) video in an integral 3-D video system. One of the problems with the EHR projection-type integral 3-D system is that positional errors appear between the elemental image and the elemental lens when there is geometric distortion in the projected image. We analyzed the relationships between the geometric distortion in the elemental images caused by the projection lens and the spatial distortion of the reconstructed 3-D image. As a result, we clarified that 3-D images reconstructed far from the lens array were greatly affected by the distortion of the elemental images, and that the 3-D images were significantly distorted in the depth direction at the corners of the displayed images. Moreover, we developed a video signal processor that electrically compensated the distortion in the elemental images for an EHR projection-type integral 3-D system. Therefore, the distortion in the displayed 3-D image was removed, and the viewing angle of the 3-D image was expanded to nearly double that obtained with the previous prototype system.

  7. PSF engineering in multifocus microscopy for increased depth volumetric imaging.

    PubMed

    Hajj, Bassam; El Beheiry, Mohamed; Dahan, Maxime

    2016-03-01

    Imaging and localizing single molecules with high accuracy in a 3D volume is a challenging task. Here we combine multifocal microscopy, a recently developed volumetric imaging technique, with point spread function engineering to achieve an increased depth for single molecule imaging. Applications in 3D single molecule localization-based super-resolution imaging is shown over an axial depth of 4 µm as well as for the tracking of diffusing beads in a fluid environment over 8 µm. PMID:27231584

  8. PSF engineering in multifocus microscopy for increased depth volumetric imaging

    PubMed Central

    Hajj, Bassam; El Beheiry, Mohamed; Dahan, Maxime

    2016-01-01

    Imaging and localizing single molecules with high accuracy in a 3D volume is a challenging task. Here we combine multifocal microscopy, a recently developed volumetric imaging technique, with point spread function engineering to achieve an increased depth for single molecule imaging. Applications in 3D single molecule localization-based super-resolution imaging is shown over an axial depth of 4 µm as well as for the tracking of diffusing beads in a fluid environment over 8 µm. PMID:27231584

  9. Post-processing methods of rendering and visualizing 3-D reconstructed tomographic images

    SciTech Connect

    Wong, S.T.C.

    1997-02-01

    The purpose of this presentation is to discuss the computer processing techniques of tomographic images, after they have been generated by imaging scanners, for volume visualization. Volume visualization is concerned with the representation, manipulation, and rendering of volumetric data. Since the first digital images were produced from computed tomography (CT) scanners in the mid 1970s, applications of visualization in medicine have expanded dramatically. Today, three-dimensional (3D) medical visualization has expanded from using CT data, the first inherently digital source of 3D medical data, to using data from various medical imaging modalities, including magnetic resonance scanners, positron emission scanners, digital ultrasound, electronic and confocal microscopy, and other medical imaging modalities. We have advanced from rendering anatomy to aid diagnosis and visualize complex anatomic structures to planning and assisting surgery and radiation treatment. New, more accurate and cost-effective procedures for clinical services and biomedical research have become possible by integrating computer graphics technology with medical images. This trend is particularly noticeable in current market-driven health care environment. For example, interventional imaging, image-guided surgery, and stereotactic and visualization techniques are now stemming into surgical practice. In this presentation, we discuss only computer-display-based approaches of volumetric medical visualization. That is, we assume that the display device available is two-dimensional (2D) in nature and all analysis of multidimensional image data is to be carried out via the 2D screen of the device. There are technologies such as holography and virtual reality that do provide a {open_quotes}true 3D screen{close_quotes}. To confine the scope, this presentation will not discuss such approaches.

  10. 3D image analysis of a volcanic deposit

    NASA Astrophysics Data System (ADS)

    de Witte, Y.; Vlassenbroeck, J.; Vandeputte, K.; Dewanckele, J.; Cnudde, V.; van Hoorebeke, L.; Ernst, G.; Jacobs, P.

    2009-04-01

    During the last decades, X-ray micro CT has become a well established technique for non-destructive testing in a wide variety of research fields. Using a series of X-ray transmission images of the sample at different projection angles, a stack of 2D cross-sections is reconstructed, resulting in a 3D volume representing the X-ray attenuation coefficients of the sample. Since the attenuation coefficient of a material depends on its density and atomic number, this volume provides valuable information about the internal structure and composition of the sample. Although much qualitative information can be derived directly from this 3D volume, researchers usually require more quantitative results to be able to provide a full characterization of the sample under investigation. This type of information needs to be retrieved using specialized image processing software. For most samples, it is imperative that this processing is performed on the 3D volume as a whole, since a sequence of 2D cross sections usually forms an inadequate approximation of the actual structure. The complete processing of a volume consists of three sequential steps. First, the volume is segmented into a set of objects. What these objects represent depends on what property of the sample needs to be analysed. The objects can be for instance concavities, dense inclusions or the matrix of the sample. When dealing with noisy data, it might be necessary to filter the data before applying the segmentation. The second step is the separation of connected objects into a set of smaller objects. This is necessary when objects appear to be connected because of the limited resolution and contrast of the scan. Separation can also be useful when the sample contains a network structure and one wants to study the individual cells of the network. The third and last step consists of the actual analysis of the various objects to derive the different parameters of interest. While some parameters require extensive

  11. 3D map of the plant photosystem II supercomplex obtained by cryoelectron microscopy and single particle analysis.

    PubMed

    Nield, J; Orlova, E V; Morris, E P; Gowen, B; van Heel, M; Barber, J

    2000-01-01

    Here we describe the first 3D structure of the photosystem II (PSII) supercomplex of higher plants, constructed by single particle analysis of images obtained by cryoelectron microscopy. This large multisubunit membrane protein complex functions to absorb light energy and catalyze the oxidation of water and reduction of plastoquinone. The resolution of the 3D structure is 24 A and emphasizes the dimeric nature of the supercomplex. The extrinsic proteins of the oxygen-evolving complex (OEC) are readily observed as a tetrameric cluster bound to the lumenal surface. By considering higher resolution data, obtained from electron crystallography, it has been possible to relate the binding sites of the OEC proteins with the underlying intrinsic membrane subunits of the photochemical reaction center core. The model suggests that the 33 kDa OEC protein is located towards the CP47/D2 side of the reaction center but is also positioned over the C-terminal helices of the D1 protein including its CD lumenal loop. In contrast, the model predicts that the 23/17 kDa OEC proteins are positioned at the N-terminus of the D1 protein incorporating the AB lumenal loop of this protein and two other unidentified transmembrane helices. Overall the 3D model represents a significant step forward in revealing the structure of the photosynthetic OEC whose activity is required to sustain the aerobic atmosphere on our planet.

  12. 3D imaging of nanomaterials by discrete tomography.

    PubMed

    Batenburg, K J; Bals, S; Sijbers, J; Kübel, C; Midgley, P A; Hernandez, J C; Kaiser, U; Encina, E R; Coronado, E A; Van Tendeloo, G

    2009-05-01

    The field of discrete tomography focuses on the reconstruction of samples that consist of only a few different materials. Ideally, a three-dimensional (3D) reconstruction of such a sample should contain only one grey level for each of the compositions in the sample. By exploiting this property in the reconstruction algorithm, either the quality of the reconstruction can be improved significantly, or the number of required projection images can be reduced. The discrete reconstruction typically contains fewer artifacts and does not have to be segmented, as it already contains one grey level for each composition. Recently, a new algorithm, called discrete algebraic reconstruction technique (DART), has been proposed that can be used effectively on experimental electron tomography datasets. In this paper, we propose discrete tomography as a general reconstruction method for electron tomography in materials science. We describe the basic principles of DART and show that it can be applied successfully to three different types of samples, consisting of embedded ErSi(2) nanocrystals, a carbon nanotube grown from a catalyst particle and a single gold nanoparticle, respectively. PMID:19269094

  13. 3D Soil Images Structure Quantification using Relative Entropy

    NASA Astrophysics Data System (ADS)

    Tarquis, A. M.; Gonzalez-Nieto, P. L.; Bird, N. R. A.

    2012-04-01

    Soil voids manifest the cumulative effect of local pedogenic processes and ultimately influence soil behavior - especially as it pertains to aeration and hydrophysical properties. Because of the relatively weak attenuation of X-rays by air, compared with liquids or solids, non-disruptive CT scanning has become a very attractive tool for generating three-dimensional imagery of soil voids. One of the main steps involved in this analysis is the thresholding required to transform the original (greyscale) images into the type of binary representation (e.g., pores in white, solids in black) needed for fractal analysis or simulation with Lattice-Boltzmann models (Baveye et al., 2010). The objective of the current work is to apply an innovative approach to quantifying soil voids and pore networks in original X-ray CT imagery using Relative Entropy (Bird et al., 2006; Tarquis et al., 2008). These will be illustrated using typical imagery representing contrasting soil structures. Particular attention will be given to the need to consider the full 3D context of the CT imagery, as well as scaling issues, in the application and interpretation of this index.

  14. A correlative approach for combining microCT, light and transmission electron microscopy in a single 3D scenario

    PubMed Central

    2013-01-01

    Background In biomedical research, a huge variety of different techniques is currently available for the structural examination of small specimens, including conventional light microscopy (LM), transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), microscopic X-ray computed tomography (microCT), and many others. Since every imaging method is physically limited by certain parameters, a correlative use of complementary methods often yields a significant broader range of information. Here we demonstrate the advantages of the correlative use of microCT, light microscopy, and transmission electron microscopy for the analysis of small biological samples. Results We used a small juvenile bivalve mollusc (Mytilus galloprovincialis, approximately 0.8 mm length) to demonstrate the workflow of a correlative examination by microCT, LM serial section analysis, and TEM-re-sectioning. Initially these three datasets were analyzed separately, and subsequently they were fused in one 3D scene. This workflow is very straightforward. The specimen was processed as usual for transmission electron microscopy including post-fixation in osmium tetroxide and embedding in epoxy resin. Subsequently it was imaged with microCT. Post-fixation in osmium tetroxide yielded sufficient X-ray contrast for microCT imaging, since the X-ray absorption of epoxy resin is low. Thereafter, the same specimen was serially sectioned for LM investigation. The serial section images were aligned and specific organ systems were reconstructed based on manual segmentation and surface rendering. According to the region of interest (ROI), specific LM sections were detached from the slides, re-mounted on resin blocks and re-sectioned (ultrathin) for TEM. For analysis, image data from the three different modalities was co-registered into a single 3D scene using the software AMIRA®. We were able to register both the LM section series volume and TEM slices neatly to the microCT dataset, with

  15. 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.

  16. Automated 3D renal segmentation based on image partitioning

    NASA Astrophysics Data System (ADS)

    Yeghiazaryan, Varduhi; Voiculescu, Irina D.

    2016-03-01

    Despite several decades of research into segmentation techniques, automated medical image segmentation is barely usable in a clinical context, and still at vast user time expense. This paper illustrates unsupervised organ segmentation through the use of a novel automated labelling approximation algorithm followed by a hypersurface front propagation method. The approximation stage relies on a pre-computed image partition forest obtained directly from CT scan data. We have implemented all procedures to operate directly on 3D volumes, rather than slice-by-slice, because our algorithms are dimensionality-independent. The results picture segmentations which identify kidneys, but can easily be extrapolated to other body parts. Quantitative analysis of our automated segmentation compared against hand-segmented gold standards indicates an average Dice similarity coefficient of 90%. Results were obtained over volumes of CT data with 9 kidneys, computing both volume-based similarity measures (such as the Dice and Jaccard coefficients, true positive volume fraction) and size-based measures (such as the relative volume difference). The analysis considered both healthy and diseased kidneys, although extreme pathological cases were excluded from the overall count. Such cases are difficult to segment both manually and automatically due to the large amplitude of Hounsfield unit distribution in the scan, and the wide spread of the tumorous tissue inside the abdomen. In the case of kidneys that have maintained their shape, the similarity range lies around the values obtained for inter-operator variability. Whilst the procedure is fully automated, our tools also provide a light level of manual editing.

  17. 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...

  18. 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...

  19. Deformable M-Reps for 3D Medical Image Segmentation

    PubMed Central

    Pizer, Stephen M.; Fletcher, P. Thomas; Joshi, Sarang; Thall, Andrew; Chen, James Z.; Fridman, Yonatan; Fritsch, Daniel S.; Gash, Graham; Glotzer, John M.; Jiroutek, Michael R.; Lu, Conglin; Muller, Keith E.; Tracton, Gregg; Yushkevich, Paul; Chaney, Edward L.

    2013-01-01

    M-reps (formerly called DSLs) are a multiscale medial means for modeling and rendering 3D solid geometry. They are particularly well suited to model anatomic objects and in particular to capture prior geometric information effectively in deformable models segmentation approaches. The representation is based on figural models, which define objects at coarse scale by a hierarchy of figures – each figure generally a slab representing a solid region and its boundary simultaneously. This paper focuses on the use of single figure models to segment objects of relatively simple structure. A single figure is a sheet of medial atoms, which is interpolated from the model formed by a net, i.e., a mesh or chain, of medial atoms (hence the name m-reps), each atom modeling a solid region via not only a position and a width but also a local figural frame giving figural directions and an object angle between opposing, corresponding positions on the boundary implied by the m-rep. The special capability of an m-rep is to provide spatial and orientational correspondence between an object in two different states of deformation. This ability is central to effective measurement of both geometric typicality and geometry to image match, the two terms of the objective function optimized in segmentation by deformable models. The other ability of m-reps central to effective segmentation is their ability to support segmentation at multiple levels of scale, with successively finer precision. Objects modeled by single figures are segmented first by a similarity transform augmented by object elongation, then by adjustment of each medial atom, and finally by displacing a dense sampling of the m-rep implied boundary. While these models and approaches also exist in 2D, we focus on 3D objects. The segmentation of the kidney from CT and the hippocampus from MRI serve as the major examples in this paper. The accuracy of segmentation as compared to manual, slice-by-slice segmentation is reported. PMID

  20. 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

  1. Multiplexed 3D FRET imaging in deep tissue of live embryos

    PubMed Central

    Zhao, Ming; Wan, Xiaoyang; Li, Yu; Zhou, Weibin; Peng, Leilei

    2015-01-01

    Current deep tissue microscopy techniques are mostly restricted to intensity mapping of fluorophores, which significantly limit their applications in investigating biochemical processes in vivo. We present a deep tissue multiplexed functional imaging method that probes multiple Förster resonant energy transfer (FRET) sensors in live embryos with high spatial resolution. The method simultaneously images fluorescence lifetimes in 3D with multiple excitation lasers. Through quantitative analysis of triple-channel intensity and lifetime images, we demonstrated that Ca2+ and cAMP levels of live embryos expressing dual FRET sensors can be monitored simultaneously at microscopic resolution. The method is compatible with a broad range of FRET sensors currently available for probing various cellular biochemical functions. It opens the door to imaging complex cellular circuitries in whole live organisms. PMID:26387920

  2. Segmentation of Image Data from Complex Organotypic 3D Models of Cancer Tissues with Markov Random Fields

    PubMed Central

    Robinson, Sean; Guyon, Laurent; Nevalainen, Jaakko; Toriseva, Mervi

    2015-01-01

    Organotypic, three dimensional (3D) cell culture models of epithelial tumour types such as prostate cancer recapitulate key aspects of the architecture and histology of solid cancers. Morphometric analysis of multicellular 3D organoids is particularly important when additional components such as the extracellular matrix and tumour microenvironment are included in the model. The complexity of such models has so far limited their successful implementation. There is a great need for automatic, accurate and robust image segmentation tools to facilitate the analysis of such biologically relevant 3D cell culture models. We present a segmentation method based on Markov random fields (MRFs) and illustrate our method using 3D stack image data from an organotypic 3D model of prostate cancer cells co-cultured with cancer-associated fibroblasts (CAFs). The 3D segmentation output suggests that these cell types are in physical contact with each other within the model, which has important implications for tumour biology. Segmentation performance is quantified using ground truth labels and we show how each step of our method increases segmentation accuracy. We provide the ground truth labels along with the image data and code. Using independent image data we show that our segmentation method is also more generally applicable to other types of cellular microscopy and not only limited to fluorescence microscopy. PMID:26630674

  3. Segmentation of Image Data from Complex Organotypic 3D Models of Cancer Tissues with Markov Random Fields.

    PubMed

    Robinson, Sean; Guyon, Laurent; Nevalainen, Jaakko; Toriseva, Mervi; Åkerfelt, Malin; Nees, Matthias

    2015-01-01

    Organotypic, three dimensional (3D) cell culture models of epithelial tumour types such as prostate cancer recapitulate key aspects of the architecture and histology of solid cancers. Morphometric analysis of multicellular 3D organoids is particularly important when additional components such as the extracellular matrix and tumour microenvironment are included in the model. The complexity of such models has so far limited their successful implementation. There is a great need for automatic, accurate and robust image segmentation tools to facilitate the analysis of such biologically relevant 3D cell culture models. We present a segmentation method based on Markov random fields (MRFs) and illustrate our method using 3D stack image data from an organotypic 3D model of prostate cancer cells co-cultured with cancer-associated fibroblasts (CAFs). The 3D segmentation output suggests that these cell types are in physical contact with each other within the model, which has important implications for tumour biology. Segmentation performance is quantified using ground truth labels and we show how each step of our method increases segmentation accuracy. We provide the ground truth labels along with the image data and code. Using independent image data we show that our segmentation method is also more generally applicable to other types of cellular microscopy and not only limited to fluorescence microscopy.

  4. Imaging and 3D Reconstruction of Cerebrovascular Structures in Embryonic Zebrafish

    PubMed Central

    Ethell, Douglas W.; Cameron, D. Joshua

    2014-01-01

    Zebrafish are a powerful tool to study developmental biology and pathology in vivo. The small size and relative transparency of zebrafish embryos make them particularly useful for the visual examination of processes such as heart and vascular development. In several recent studies transgenic zebrafish that express EGFP in vascular endothelial cells were used to image and analyze complex vascular networks in the brain and retina, using confocal microscopy. Descriptions are provided to prepare, treat and image zebrafish embryos that express enhanced green fluorescent protein (EGFP), and then generate comprehensive 3D renderings of the cerebrovascular system. Protocols include the treatment of embryos, confocal imaging, and fixation protocols that preserve EGFP fluorescence. Further, useful tips on obtaining high-quality images of cerebrovascular structures, such as removal the eye without damaging nearby neural tissue are provided. Potential pitfalls with confocal imaging are discussed, along with the steps necessary to generate 3D reconstructions from confocal image stacks using freely available open source software. PMID:24797110

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

    PubMed Central

    Craig, Roger

    2011-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  7. 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.

  8. Characterization and application of 3D-printed phantoms for biophotonic imaging

    NASA Astrophysics Data System (ADS)

    Wang, Jianting; Coburn, James; Liang, Chia-Pin; Woolsey, Nicholas; Le, Du; Ramella-Roman, Jessica; Chen, Yu; Pfefer, Joshua

    2013-05-01

    The emerging technique of three-dimensional (3D) printing provides a simple, fast, and flexible way to fabricate structures with arbitrary spatial features and may prove useful in the development of standardized, phantom-based performance test methods for biophotonic imaging. Acrylonitrile Butadiene Styrene (ABS) is commonly used in the printing process, given its low cost and strength. In this study, we evaluate 3D printing as an approach for fabricating biologically-relevant optical phantoms for hyperspectral reflectance imaging (HRI). The initial phase of this work involved characterization of absorption and scattering coefficients using spectrophotometry. The morphology of phantoms incorporating vessel-like channels with diameters on the order of hundreds of microns was examined by microscopy and OCT. A near-infrared absorbing dye was injected into channels located at a range of depths within the phantom and imaged with a near-infrared HRI system (650-1100 nm). ABS was found to have scattering coefficients comparable to biological tissue and low absorption throughout much of the visible and infrared range. Channels with dimensions on the order of the resolution limit of the 3D printer (~0.2 mm) exhibited pixelation effects as well as a degree of distortion along their edges. Furthermore, phantom porosity sometimes resulted in leakage from channel regions. Contrast-enhanced channel visualization with HRI was possible to a depth of nearly 1 mm - a level similar to that seen previously in biological tissue. Overall, our ABS phantoms demonstrated a high level of optical similarity to biological tissue. While limitations in printer resolution, matrix homogeneity and optical property tunability remain challenging, 3D printed phantoms have significant promise as samples for objective, quantitative evaluation of performance for biophotonic imaging modalities such as HRI.

  9. Assessing 3D tunnel position in ACL reconstruction using a novel single image 3D-2D registration

    NASA Astrophysics Data System (ADS)

    Kang, X.; Yau, W. P.; Otake, Y.; Cheung, P. Y. S.; Hu, Y.; Taylor, R. H.

    2012-02-01

    The routinely used procedure for evaluating tunnel positions following anterior cruciate ligament (ACL) reconstructions based on standard X-ray images is known to pose difficulties in terms of obtaining accurate measures, especially in providing three-dimensional tunnel positions. This is largely due to the variability in individual knee joint pose relative to X-ray plates. Accurate results were reported using postoperative CT. However, its extensive usage in clinical routine is hampered by its major requirement of having CT scans of individual patients, which is not available for most ACL reconstructions. These difficulties are addressed through the proposed method, which aligns a knee model to X-ray images using our novel single-image 3D-2D registration method and then estimates the 3D tunnel position. In the proposed method, the alignment is achieved by using a novel contour-based 3D-2D registration method wherein image contours are treated as a set of oriented points. However, instead of using some form of orientation weighting function and multiplying it with a distance function, we formulate the 3D-2D registration as a probability density estimation using a mixture of von Mises-Fisher-Gaussian (vMFG) distributions and solve it through an expectation maximization (EM) algorithm. Compared with the ground-truth established from postoperative CT, our registration method in an experiment using a plastic phantom showed accurate results with errors of (-0.43°+/-1.19°, 0.45°+/-2.17°, 0.23°+/-1.05°) and (0.03+/-0.55, -0.03+/-0.54, -2.73+/-1.64) mm. As for the entry point of the ACL tunnel, one of the key measurements, it was obtained with high accuracy of 0.53+/-0.30 mm distance errors.

  10. 3-D Adaptive Sparsity Based Image Compression with Applications to Optical Coherence Tomography

    PubMed Central

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

    2015-01-01

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

  11. Segmentation of densely populated cell nuclei from confocal image stacks using 3D non-parametric shape priors.

    PubMed

    Ong, Lee-Ling S; Wang, Mengmeng; Dauwels, Justin; Asada, H Harry

    2014-01-01

    An approach to jointly estimate 3D shapes and poses of stained nuclei from confocal microscopy images, using statistical prior information, is presented. Extracting nuclei boundaries from our experimental images of cell migration is challenging due to clustered nuclei and variations in their shapes. This issue is formulated as a maximum a posteriori estimation problem. By incorporating statistical prior models of 3D nuclei shapes into level set functions, the active contour evolutions applied on the images is constrained. A 3D alignment algorithm is developed to build the training databases and to match contours obtained from the images to them. To address the issue of aligning the model over multiple clustered nuclei, a watershed-like technique is used to detect and separate clustered regions prior to active contour evolution. Our method is tested on confocal images of endothelial cells in microfluidic devices, compared with existing approaches.

  12. 3-D Imaging Systems for Agricultural Applications-A Review.

    PubMed

    Vázquez-Arellano, Manuel; Griepentrog, Hans W; Reiser, David; Paraforos, Dimitris S

    2016-01-01

    Efficiency increase of resources through automation of agriculture requires more information about the production process, as well as process and machinery status. Sensors are necessary for monitoring the status and condition of production by recognizing the surrounding structures such as objects, field structures, natural or artificial markers, and obstacles. Currently, three dimensional (3-D) sensors are economically affordable and technologically advanced to a great extent, so a breakthrough is already possible if enough research projects are commercialized. The aim of this review paper is to investigate the state-of-the-art of 3-D vision systems in agriculture, and the role and value that only 3-D data can have to provide information about environmental structures based on the recent progress in optical 3-D sensors. The structure of this research consists of an overview of the different optical 3-D vision techniques, based on the basic principles. Afterwards, their application in agriculture are reviewed. The main focus lays on vehicle navigation, and crop and animal husbandry. The depth dimension brought by 3-D sensors provides key information that greatly facilitates the implementation of automation and robotics in agriculture.

  13. 3-D Imaging Systems for Agricultural Applications-A Review.

    PubMed

    Vázquez-Arellano, Manuel; Griepentrog, Hans W; Reiser, David; Paraforos, Dimitris S

    2016-01-01

    Efficiency increase of resources through automation of agriculture requires more information about the production process, as well as process and machinery status. Sensors are necessary for monitoring the status and condition of production by recognizing the surrounding structures such as objects, field structures, natural or artificial markers, and obstacles. Currently, three dimensional (3-D) sensors are economically affordable and technologically advanced to a great extent, so a breakthrough is already possible if enough research projects are commercialized. The aim of this review paper is to investigate the state-of-the-art of 3-D vision systems in agriculture, and the role and value that only 3-D data can have to provide information about environmental structures based on the recent progress in optical 3-D sensors. The structure of this research consists of an overview of the different optical 3-D vision techniques, based on the basic principles. Afterwards, their application in agriculture are reviewed. The main focus lays on vehicle navigation, and crop and animal husbandry. The depth dimension brought by 3-D sensors provides key information that greatly facilitates the implementation of automation and robotics in agriculture. PMID:27136560

  14. 3-D Imaging Systems for Agricultural Applications—A Review

    PubMed Central

    Vázquez-Arellano, Manuel; Griepentrog, Hans W.; Reiser, David; Paraforos, Dimitris S.

    2016-01-01

    Efficiency increase of resources through automation of agriculture requires more information about the production process, as well as process and machinery status. Sensors are necessary for monitoring the status and condition of production by recognizing the surrounding structures such as objects, field structures, natural or artificial markers, and obstacles. Currently, three dimensional (3-D) sensors are economically affordable and technologically advanced to a great extent, so a breakthrough is already possible if enough research projects are commercialized. The aim of this review paper is to investigate the state-of-the-art of 3-D vision systems in agriculture, and the role and value that only 3-D data can have to provide information about environmental structures based on the recent progress in optical 3-D sensors. The structure of this research consists of an overview of the different optical 3-D vision techniques, based on the basic principles. Afterwards, their application in agriculture are reviewed. The main focus lays on vehicle navigation, and crop and animal husbandry. The depth dimension brought by 3-D sensors provides key information that greatly facilitates the implementation of automation and robotics in agriculture. PMID:27136560

  15. 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.

  16. Dense 3d Point Cloud Generation from Uav Images from Image Matching and Global Optimazation

    NASA Astrophysics Data System (ADS)

    Rhee, S.; Kim, T.

    2016-06-01

    3D spatial information from unmanned aerial vehicles (UAV) images is usually provided in the form of 3D point clouds. For various UAV applications, it is important to generate dense 3D point clouds automatically from over the entire extent of UAV images. In this paper, we aim to apply image matching for generation of local point clouds over a pair or group of images and global optimization to combine local point clouds over the whole region of interest. We tried to apply two types of image matching, an object space-based matching technique and an image space-based matching technique, and to compare the performance of the two techniques. The object space-based matching used here sets a list of candidate height values for a fixed horizontal position in the object space. For each height, its corresponding image point is calculated and similarity is measured by grey-level correlation. The image space-based matching used here is a modified relaxation matching. We devised a global optimization scheme for finding optimal pairs (or groups) to apply image matching, defining local match region in image- or object- space, and merging local point clouds into a global one. For optimal pair selection, tiepoints among images were extracted and stereo coverage network was defined by forming a maximum spanning tree using the tiepoints. From experiments, we confirmed that through image matching and global optimization, 3D point clouds were generated successfully. However, results also revealed some limitations. In case of image-based matching results, we observed some blanks in 3D point clouds. In case of object space-based matching results, we observed more blunders than image-based matching ones and noisy local height variations. We suspect these might be due to inaccurate orientation parameters. The work in this paper is still ongoing. We will further test our approach with more precise orientation parameters.

  17. Segmented images and 3D images for studying the anatomical structures in MRIs

    NASA Astrophysics Data System (ADS)

    Lee, Yong Sook; Chung, Min Suk; Cho, Jae Hyun

    2004-05-01

    For identifying the pathological findings in MRIs, the anatomical structures in MRIs should be identified in advance. For studying the anatomical structures in MRIs, an education al tool that includes the horizontal, coronal, sagittal MRIs of entire body, corresponding segmented images, 3D images, and browsing software is necessary. Such an educational tool, however, is hard to obtain. Therefore, in this research, such an educational tool which helps medical students and doctors study the anatomical structures in MRIs was made as follows. A healthy, young Korean male adult with standard body shape was selected. Six hundred thirteen horizontal MRIs of the entire body were scanned and inputted to the personal computer. Sixty anatomical structures in the horizontal MRIs were segmented to make horizontal segmented images. Coronal, sagittal MRIs and coronal, sagittal segmented images were made. 3D images of anatomical structures in the segmented images were reconstructed by surface rendering method. Browsing software of the MRIs, segmented images, and 3D images was composed. This educational tool that includes horizontal, coronal, sagittal MRIs of entire body, corresponding segmented images, 3D images, and browsing software is expected to help medical students and doctors study anatomical structures in MRIs.

  18. 3-D Reconstruction From 2-D Radiographic Images and Its Application to Clinical Veterinary Medicine

    NASA Astrophysics Data System (ADS)

    Hamamoto, Kazuhiko; Sato, Motoyoshi

    3D imaging technique is very important and indispensable in diagnosis. The main stream of the technique is one in which 3D image is reconstructed from a set of slice images, such as X-ray CT and MRI. However, these systems require large space and high costs. On the other hand, a low cost and small size 3D imaging system is needed in clinical veterinary medicine, for example, in the case of diagnosis in X-ray car or pasture area. We propose a novel 3D imaging technique using 2-D X-ray radiographic images. This system can be realized by cheaper system than X-ray CT and enables to get 3D image in X-ray car or portable X-ray equipment. In this paper, a 3D visualization technique from 2-D radiographic images is proposed and several reconstructions are shown. These reconstructions are evaluated by veterinarians.

  19. 3D simulation of the image formation in soft x-ray microscopes.

    PubMed

    Selin, Mårten; Fogelqvist, Emelie; Holmberg, Anders; Guttmann, Peter; Vogt, Ulrich; Hertz, Hans M

    2014-12-15

    In water-window soft x-ray microscopy the studied object is typically larger than the depth of focus and the sample illumination is often partially coherent. This blurs out-of-focus features and may introduce considerable fringing. Understanding the influence of these phenomena on the image formation is therefore important when interpreting experimental data. Here we present a wave-propagation model operating in 3D for simulating the image formation of thick objects in partially coherent soft x-ray microscopes. The model is compared with present simulation methods as well as with experiments. The results show that our model predicts the image formation of transmission soft x-ray microscopes more accurately than previous models.

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

    PubMed

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

    2014-03-25

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

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

    PubMed

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

    2016-02-23

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

  2. Dual-view integral imaging 3D display using polarizer parallax barriers.

    PubMed

    Wu, Fei; Wang, Qiong-Hua; Luo, Cheng-Gao; Li, Da-Hai; Deng, Huan

    2014-04-01

    We propose a dual-view integral imaging (DVII) 3D display using polarizer parallax barriers (PPBs). The DVII 3D display consists of a display panel, a microlens array, and two PPBs. The elemental images (EIs) displayed on the left and right half of the display panel are captured from two different 3D scenes, respectively. The lights emitted from two kinds of EIs are modulated by the left and right half of the microlens array to present two different 3D images, respectively. A prototype of the DVII 3D display is developed, and the experimental results agree well with the theory.

  3. Free segmentation in rendered 3D images through synthetic impulse response in integral imaging

    NASA Astrophysics Data System (ADS)

    Martínez-Corral, M.; Llavador, A.; Sánchez-Ortiga, E.; Saavedra, G.; Javidi, B.

    2016-06-01

    Integral Imaging is a technique that has the capability of providing not only the spatial, but also the angular information of three-dimensional (3D) scenes. Some important applications are the 3D display and digital post-processing as for example, depth-reconstruction from integral images. In this contribution we propose a new reconstruction method that takes into account the integral image and a simplified version of the impulse response function (IRF) of the integral imaging (InI) system to perform a two-dimensional (2D) deconvolution. The IRF of an InI system has a periodic structure that depends directly on the axial position of the object. Considering different periods of the IRFs we recover by deconvolution the depth information of the 3D scene. An advantage of our method is that it is possible to obtain nonconventional reconstructions by considering alternative synthetic impulse responses. Our experiments show the feasibility of the proposed method.

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

    PubMed

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

    2012-05-01

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

  5. Implementation of wireless 3D stereo image capture system and 3D exaggeration algorithm for the region of interest

    NASA Astrophysics Data System (ADS)

    Ham, Woonchul; Song, Chulgyu; Lee, Kangsan; Badarch, Luubaatar

    2015-05-01

    In this paper, we introduce the mobile embedded system implemented for capturing stereo image based on two CMOS camera module. We use WinCE as an operating system and capture the stereo image by using device driver for CMOS camera interface and Direct Draw API functions. We aslo comments on the GPU hardware and CUDA programming for implementation of 3D exaggeraion algorithm for ROI by adjusting and synthesizing the disparity value of ROI (region of interest) in real time. We comment on the pattern of aperture for deblurring of CMOS camera module based on the Kirchhoff diffraction formula and clarify the reason why we can get more sharp and clear image by blocking some portion of aperture or geometric sampling. Synthesized stereo image is real time monitored on the shutter glass type three-dimensional LCD monitor and disparity values of each segment are analyzed to prove the validness of emphasizing effect of ROI.

  6. Imaging 3D strain field monitoring during hydraulic fracturing processes

    NASA Astrophysics Data System (ADS)

    Chen, Rongzhang; Zaghloul, Mohamed A. S.; Yan, Aidong; Li, Shuo; Lu, Guanyi; Ames, Brandon C.; Zolfaghari, Navid; Bunger, Andrew P.; Li, Ming-Jun; Chen, Kevin P.

    2016-05-01

    In this paper, we present a distributed fiber optic sensing scheme to study 3D strain fields inside concrete cubes during hydraulic fracturing process. Optical fibers embedded in concrete were used to monitor 3D strain field build-up with external hydraulic pressures. High spatial resolution strain fields were interrogated by the in-fiber Rayleigh backscattering with 1-cm spatial resolution using optical frequency domain reflectometry. The fiber optics sensor scheme presented in this paper provides scientists and engineers a unique laboratory tool to understand the hydraulic fracturing processes in various rock formations and its impacts to environments.

  7. 3D fingerprint imaging system based on full-field fringe projection profilometry

    NASA Astrophysics Data System (ADS)

    Huang, Shujun; Zhang, Zonghua; Zhao, Yan; Dai, Jie; Chen, Chao; Xu, Yongjia; Zhang, E.; Xie, Lili

    2014-01-01

    As an unique, unchangeable and easily acquired biometrics, fingerprint has been widely studied in academics and applied in many fields over the years. The traditional fingerprint recognition methods are based on the obtained 2D feature of fingerprint. However, fingerprint is a 3D biological characteristic. The mapping from 3D to 2D loses 1D information and causes nonlinear distortion of the captured fingerprint. Therefore, it is becoming more and more important to obtain 3D fingerprint information for recognition. In this paper, a novel 3D fingerprint imaging system is presented based on fringe projection technique to obtain 3D features and the corresponding color texture information. A series of color sinusoidal fringe patterns with optimum three-fringe numbers are projected onto a finger surface. From another viewpoint, the fringe patterns are deformed by the finger surface and captured by a CCD camera. 3D shape data of the finger can be obtained from the captured fringe pattern images. This paper studies the prototype of the 3D fingerprint imaging system, including principle of 3D fingerprint acquisition, hardware design of the 3D imaging system, 3D calibration of the system, and software development. Some experiments are carried out by acquiring several 3D fingerprint data. The experimental results demonstrate the feasibility of the proposed 3D fingerprint imaging system.

  8. Image-based RSA: Roentgen stereophotogrammetric analysis based on 2D-3D image registration.

    PubMed

    de Bruin, P W; Kaptein, B L; Stoel, B C; Reiber, J H C; Rozing, P M; Valstar, E R

    2008-01-01

    Image-based Roentgen stereophotogrammetric analysis (IBRSA) integrates 2D-3D image registration and conventional RSA. Instead of radiopaque RSA bone markers, IBRSA uses 3D CT data, from which digitally reconstructed radiographs (DRRs) are generated. Using 2D-3D image registration, the 3D pose of the CT is iteratively adjusted such that the generated DRRs resemble the 2D RSA images as closely as possible, according to an image matching metric. Effectively, by registering all 2D follow-up moments to the same 3D CT, the CT volume functions as common ground. In two experiments, using RSA and using a micromanipulator as gold standard, IBRSA has been validated on cadaveric and sawbone scapula radiographs, and good matching results have been achieved. The accuracy was: |mu |< 0.083 mm for translations and |mu| < 0.023 degrees for rotations. The precision sigma in x-, y-, and z-direction was 0.090, 0.077, and 0.220 mm for translations and 0.155 degrees , 0.243 degrees , and 0.074 degrees for rotations. Our results show that the accuracy and precision of in vitro IBRSA, performed under ideal laboratory conditions, are lower than in vitro standard RSA but higher than in vivo standard RSA. Because IBRSA does not require radiopaque markers, it adds functionality to the RSA method by opening new directions and possibilities for research, such as dynamic analyses using fluoroscopy on subjects without markers and computer navigation applications.

  9. Display of travelling 3D scenes from single integral-imaging capture

    NASA Astrophysics Data System (ADS)

    Martinez-Corral, Manuel; Dorado, Adrian; Hong, Seok-Min; Sola-Pikabea, Jorge; Saavedra, Genaro

    2016-06-01

    Integral imaging (InI) is a 3D auto-stereoscopic technique that captures and displays 3D images. We present a method for easily projecting the information recorded with this technique by transforming the integral image into a plenoptic image, as well as choosing, at will, the field of view (FOV) and the focused plane of the displayed plenoptic image. Furthermore, with this method we can generate a sequence of images that simulates a camera travelling through the scene from a single integral image. The application of this method permits to improve the quality of 3D display images and videos.

  10. Estimating Density Gradients and Drivers from 3D Ionospheric Imaging

    NASA Astrophysics Data System (ADS)

    Datta-Barua, S.; Bust, G. S.; Curtis, N.; Reynolds, A.; Crowley, G.

    2009-12-01

    The transition regions at the edges of the ionospheric storm-enhanced density (SED) are important for a detailed understanding of the mid-latitude physical processes occurring during major magnetic storms. At the boundary, the density gradients are evidence of the drivers that link the larger processes of the SED, with its connection to the plasmasphere and prompt-penetration electric fields, to the smaller irregularities that result in scintillations. For this reason, we present our estimates of both the plasma variation with horizontal and vertical spatial scale of 10 - 100 km and the plasma motion within and along the edges of the SED. To estimate the density gradients, we use Ionospheric Data Assimilation Four-Dimensional (IDA4D), a mature data assimilation algorithm that has been developed over several years and applied to investigations of polar cap patches and space weather storms [Bust and Crowley, 2007; Bust et al., 2007]. We use the density specification produced by IDA4D with a new tool for deducing ionospheric drivers from 3D time-evolving electron density maps, called Estimating Model Parameters from Ionospheric Reverse Engineering (EMPIRE). The EMPIRE technique has been tested on simulated data from TIMEGCM-ASPEN and on IDA4D-based density estimates with ongoing validation from Arecibo ISR measurements [Datta-Barua et al., 2009a; 2009b]. We investigate the SED that formed during the geomagnetic super storm of November 20, 2003. We run IDA4D at low-resolution continent-wide, and then re-run it at high (~10 km horizontal and ~5-20 km vertical) resolution locally along the boundary of the SED, where density gradients are expected to be highest. We input the high-resolution estimates of electron density to EMPIRE to estimate the ExB drifts and field-aligned plasma velocities along the boundaries of the SED. We expect that these drivers contribute to the density structuring observed along the SED during the storm. Bust, G. S. and G. Crowley (2007

  11. Monopulse radar 3-D imaging and application in terminal guidance radar

    NASA Astrophysics Data System (ADS)

    Xu, Hui; Qin, Guodong; Zhang, Lina

    2007-11-01

    Monopulse radar 3-D imaging integrates ISAR, monopulse angle measurement and 3-D imaging processing to obtain the 3-D image which can reflect the real size of a target, which means any two of the three measurement parameters, namely azimuth difference beam elevation difference beam and radial range, can be used to form 3-D image of 3-D object. The basic principles of Monopulse radar 3-D imaging are briefly introduced, the effect of target carriage changes(including yaw, pitch, roll and movement of target itself) on 3-D imaging and 3-D moving compensation based on the chirp rate μ and Doppler frequency f d are analyzed, and the application of monopulse radar 3-D imaging to terminal guidance radars is forecasted. The computer simulation results show that monopulse radar 3-D imaging has apparent advantages in distinguishing a target from overside interference and precise assault on vital part of a target, and has great importance in terminal guidance radars.

  12. 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.

  13. Recognition Accuracy Using 3D Endoscopic Images for Superficial Gastrointestinal Cancer: A Crossover Study

    PubMed Central

    Kaise, Mitsuru; Kikuchi, Daisuke; Iizuka, Toshiro; Fukuma, Yumiko; Kuribayashi, Yasutaka; Tanaka, Masami; Toba, Takahito; Furuhata, Tsukasa; Yamashita, Satoshi; Matsui, Akira; Mitani, Toshifumi; Hoteya, Shu

    2016-01-01

    Aim. To determine whether 3D endoscopic images improved recognition accuracy for superficial gastrointestinal cancer compared with 2D images. Methods. We created an image catalog using 2D and 3D images of 20 specimens resected by endoscopic submucosal dissection. The twelve participants were allocated into two groups. Group 1 evaluated only 2D images at first, group 2 evaluated 3D images, and, after an interval of 2 weeks, group 1 next evaluated 3D and group 2 evaluated 2D images. The evaluation items were as follows: (1) diagnostic accuracy of the tumor extent and (2) confidence levels in assessing (a) tumor extent, (b) morphology, (c) microsurface structure, and (d) comprehensive recognition. Results. The use of 3D images resulted in an improvement in diagnostic accuracy in both group 1 (2D: 76.9%, 3D: 78.6%) and group 2 (2D: 79.9%, 3D: 83.6%), with no statistically significant difference. The confidence levels were higher for all items ((a) to (d)) when 3D images were used. With respect to experience, the degree of the improvement showed the following trend: novices > trainees > experts. Conclusions. By conversion into 3D images, there was a significant improvement in the diagnostic confidence level for superficial tumors, and the improvement was greater in individuals with lower endoscopic expertise. PMID:27597863

  14. Recognition Accuracy Using 3D Endoscopic Images for Superficial Gastrointestinal Cancer: A Crossover Study

    PubMed Central

    Kaise, Mitsuru; Kikuchi, Daisuke; Iizuka, Toshiro; Fukuma, Yumiko; Kuribayashi, Yasutaka; Tanaka, Masami; Toba, Takahito; Furuhata, Tsukasa; Yamashita, Satoshi; Matsui, Akira; Mitani, Toshifumi; Hoteya, Shu

    2016-01-01

    Aim. To determine whether 3D endoscopic images improved recognition accuracy for superficial gastrointestinal cancer compared with 2D images. Methods. We created an image catalog using 2D and 3D images of 20 specimens resected by endoscopic submucosal dissection. The twelve participants were allocated into two groups. Group 1 evaluated only 2D images at first, group 2 evaluated 3D images, and, after an interval of 2 weeks, group 1 next evaluated 3D and group 2 evaluated 2D images. The evaluation items were as follows: (1) diagnostic accuracy of the tumor extent and (2) confidence levels in assessing (a) tumor extent, (b) morphology, (c) microsurface structure, and (d) comprehensive recognition. Results. The use of 3D images resulted in an improvement in diagnostic accuracy in both group 1 (2D: 76.9%, 3D: 78.6%) and group 2 (2D: 79.9%, 3D: 83.6%), with no statistically significant difference. The confidence levels were higher for all items ((a) to (d)) when 3D images were used. With respect to experience, the degree of the improvement showed the following trend: novices > trainees > experts. Conclusions. By conversion into 3D images, there was a significant improvement in the diagnostic confidence level for superficial tumors, and the improvement was greater in individuals with lower endoscopic expertise.

  15. Recognition Accuracy Using 3D Endoscopic Images for Superficial Gastrointestinal Cancer: A Crossover Study.

    PubMed

    Nomura, Kosuke; Kaise, Mitsuru; Kikuchi, Daisuke; Iizuka, Toshiro; Fukuma, Yumiko; Kuribayashi, Yasutaka; Tanaka, Masami; Toba, Takahito; Furuhata, Tsukasa; Yamashita, Satoshi; Matsui, Akira; Mitani, Toshifumi; Hoteya, Shu

    2016-01-01

    Aim. To determine whether 3D endoscopic images improved recognition accuracy for superficial gastrointestinal cancer compared with 2D images. Methods. We created an image catalog using 2D and 3D images of 20 specimens resected by endoscopic submucosal dissection. The twelve participants were allocated into two groups. Group 1 evaluated only 2D images at first, group 2 evaluated 3D images, and, after an interval of 2 weeks, group 1 next evaluated 3D and group 2 evaluated 2D images. The evaluation items were as follows: (1) diagnostic accuracy of the tumor extent and (2) confidence levels in assessing (a) tumor extent, (b) morphology, (c) microsurface structure, and (d) comprehensive recognition. Results. The use of 3D images resulted in an improvement in diagnostic accuracy in both group 1 (2D: 76.9%, 3D: 78.6%) and group 2 (2D: 79.9%, 3D: 83.6%), with no statistically significant difference. The confidence levels were higher for all items ((a) to (d)) when 3D images were used. With respect to experience, the degree of the improvement showed the following trend: novices > trainees > experts. Conclusions. By conversion into 3D images, there was a significant improvement in the diagnostic confidence level for superficial tumors, and the improvement was greater in individuals with lower endoscopic expertise.

  16. Recognition Accuracy Using 3D Endoscopic Images for Superficial Gastrointestinal Cancer: A Crossover Study.

    PubMed

    Nomura, Kosuke; Kaise, Mitsuru; Kikuchi, Daisuke; Iizuka, Toshiro; Fukuma, Yumiko; Kuribayashi, Yasutaka; Tanaka, Masami; Toba, Takahito; Furuhata, Tsukasa; Yamashita, Satoshi; Matsui, Akira; Mitani, Toshifumi; Hoteya, Shu

    2016-01-01

    Aim. To determine whether 3D endoscopic images improved recognition accuracy for superficial gastrointestinal cancer compared with 2D images. Methods. We created an image catalog using 2D and 3D images of 20 specimens resected by endoscopic submucosal dissection. The twelve participants were allocated into two groups. Group 1 evaluated only 2D images at first, group 2 evaluated 3D images, and, after an interval of 2 weeks, group 1 next evaluated 3D and group 2 evaluated 2D images. The evaluation items were as follows: (1) diagnostic accuracy of the tumor extent and (2) confidence levels in assessing (a) tumor extent, (b) morphology, (c) microsurface structure, and (d) comprehensive recognition. Results. The use of 3D images resulted in an improvement in diagnostic accuracy in both group 1 (2D: 76.9%, 3D: 78.6%) and group 2 (2D: 79.9%, 3D: 83.6%), with no statistically significant difference. The confidence levels were higher for all items ((a) to (d)) when 3D images were used. With respect to experience, the degree of the improvement showed the following trend: novices > trainees > experts. Conclusions. By conversion into 3D images, there was a significant improvement in the diagnostic confidence level for superficial tumors, and the improvement was greater in individuals with lower endoscopic expertise. PMID:27597863

  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. Correction of depth-dependent aberrations in 3D single-molecule localization and super-resolution microscopy.

    PubMed

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

    2014-01-15

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

  19. Stimulated Raman scattering microscopy for biomedical imaging

    NASA Astrophysics Data System (ADS)

    Min, Wei; Freudiger, Christian W.; Lu, Sijia; He, Chengwei; Kang, Jing X.; Xie, X. Sunney

    2009-02-01

    Label-free chemical contrast is highly desirable in biomedical imaging. Spontaneous Raman microscopy provides specific vibrational signatures of chemical bonds, but is often hindered by low sensitivity. Here we report a 3D multi-photon vibrational imaging technique based on stimulated Raman scattering (SRS). The sensitivity of SRS is significantly greater than that of spontaneous Raman scattering, and is further enhanced by high-frequency (MHz) phase-sensitive detection. SRS microscopy has a major advantage over previous coherent Raman techniques in that it offers background-free and easily interpretable chemical contrast. We show a variety of biomedical applications, such as differentiating distributions of omega-3 fatty acids and saturated lipids in living cells, imaging of brain and skin tissues based on intrinsic lipid contrast.

  20. 360 degree realistic 3D image display and image processing from real objects

    NASA Astrophysics Data System (ADS)

    Luo, Xin; Chen, Yue; Huang, Yong; Tan, Xiaodi; Horimai, Hideyoshi

    2016-09-01

    A 360-degree realistic 3D image display system based on direct light scanning method, so-called Holo-Table has been introduced in this paper. High-density directional continuous 3D motion images can be displayed easily with only one spatial light modulator. Using the holographic screen as the beam deflector, 360-degree full horizontal viewing angle was achieved. As an accompany part of the system, CMOS camera based image acquisition platform was built to feed the display engine, which can take a full 360-degree continuous imaging of the sample at the center. Customized image processing techniques such as scaling, rotation, format transformation were also developed and embedded into the system control software platform. In the end several samples were imaged to demonstrate the capability of our system.

  1. Accuracy of volume measurement using 3D ultrasound and development of CT-3D US image fusion algorithm for prostate cancer radiotherapy

    SciTech Connect

    Baek, Jihye; Huh, Jangyoung; Hyun An, So; Oh, Yoonjin; Kim, Myungsoo; Kim, DongYoung; Chung, Kwangzoo; Cho, Sungho; Lee, Rena

    2013-02-15

    Purpose: To evaluate the accuracy of measuring volumes using three-dimensional ultrasound (3D US), and to verify the feasibility of the replacement of CT-MR fusion images with CT-3D US in radiotherapy treatment planning. Methods: Phantoms, consisting of water, contrast agent, and agarose, were manufactured. The volume was measured using 3D US, CT, and MR devices. A CT-3D US and MR-3D US image fusion software was developed using the Insight Toolkit library in order to acquire three-dimensional fusion images. The quality of the image fusion was evaluated using metric value and fusion images. Results: Volume measurement, using 3D US, shows a 2.8 {+-} 1.5% error, 4.4 {+-} 3.0% error for CT, and 3.1 {+-} 2.0% error for MR. The results imply that volume measurement using the 3D US devices has a similar accuracy level to that of CT and MR. Three-dimensional image fusion of CT-3D US and MR-3D US was successfully performed using phantom images. Moreover, MR-3D US image fusion was performed using human bladder images. Conclusions: 3D US could be used in the volume measurement of human bladders and prostates. CT-3D US image fusion could be used in monitoring the target position in each fraction of external beam radiation therapy. Moreover, the feasibility of replacing the CT-MR image fusion to the CT-3D US in radiotherapy treatment planning was verified.

  2. Image enhancement and segmentation of fluid-filled structures in 3D ultrasound images

    NASA Astrophysics Data System (ADS)

    Chalana, Vikram; Dudycha, Stephen; McMorrow, Gerald

    2003-05-01

    Segmentation of fluid-filled structures, such as the urinary bladder, from three-dimensional ultrasound images is necessary for measuring their volume. This paper describes a system for image enhancement, segmentation and volume measurement of fluid-filled structures on 3D ultrasound images. The system was applied for the measurement of urinary bladder volume. Results show an average error of less than 10% in the estimation of the total bladder volume.

  3. Estimation of single cell volume from 3D confocal images using automatic data processing

    NASA Astrophysics Data System (ADS)

    Chorvatova, A.; Cagalinec, M.; Mateasik, A.; Chorvat, D., Jr.

    2012-06-01

    Cardiac cells are highly structured with a non-uniform morphology. Although precise estimation of their volume is essential for correct evaluation of hypertrophic changes of the heart, simple and unified techniques that allow determination of the single cardiomyocyte volume with sufficient precision are still limited. Here, we describe a novel approach to assess the cell volume from confocal microscopy 3D images of living cardiac myocytes. We propose a fast procedure based on segementation using active deformable contours. This technique is independent on laser gain and/or pinhole settings and it is also applicable on images of cells stained with low fluorescence markers. Presented approach is a promising new tool to investigate changes in the cell volume during normal, as well as pathological growth, as we demonstrate in the case of cell enlargement during hypertension in rats.

  4. Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging.

    PubMed

    Kumar, S; Dunsby, C; De Beule, P A A; Owen, D M; Anand, U; Lanigan, P M P; Benninger, R K P; Davis, D M; Neil, M A A; Anand, P; Benham, C; Naylor, A; French, P M W

    2007-10-01

    We report a multifocal multiphoton time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) microscope system that uses a 16 channel multi-anode PMT detector. Multiphoton excitation minimizes out-of-focus photobleaching, multifocal excitation reduces non-linear in-plane photobleaching effects and TCSPC electronics provide photon-efficient detection of the fluorescence decay profile. TCSPC detection is less prone to bleaching- and movement-induced artefacts compared to wide-field time-gated or frequency-domain FLIM. This microscope is therefore capable of acquiring 3-D FLIM images at significantly increased speeds compared to single beam multiphoton microscopy and we demonstrate this with live cells expressing a GFP tagged protein. We also apply this system to time-lapse FLIM of NAD(P)H autofluorescence in single live cells and report measurements on the change in the fluorescence decay profile following the application of a known metabolic inhibitor. PMID:19550524

  5. 3D Image Reconstructions and the Nyquist-Shannon Theorem

    NASA Astrophysics Data System (ADS)

    Ficker, T.; Martišek, D.

    2015-09-01

    Fracture surfaces are occasionally modelled by Fourier's two-dimensional series that can be converted into digital 3D reliefs mapping the morphology of solid surfaces. Such digital replicas may suffer from various artefacts when processed inconveniently. Spatial aliasing is one of those artefacts that may devalue Fourier's replicas. According to the Nyquist-Shannon sampling theorem the spatial aliasing occurs when Fourier's frequencies exceed the Nyquist critical frequency. In the present paper it is shown that the Nyquist frequency is not the only critical limit determining aliasing artefacts but there are some other frequencies that intensify aliasing phenomena and form an infinite set of points at which numerical results abruptly and dramatically change their values. This unusual type of spatial aliasing is explored and some consequences for 3D computer reconstructions are presented.

  6. Computation of optimized arrays for 3-D electrical imaging surveys

    NASA Astrophysics Data System (ADS)

    Loke, M. H.; Wilkinson, P. B.; Uhlemann, S. S.; Chambers, J. E.; Oxby, L. S.

    2014-12-01

    3-D electrical resistivity surveys and inversion models are required to accurately resolve structures in areas with very complex geology where 2-D models might suffer from artefacts. Many 3-D surveys use a grid where the number of electrodes along one direction (x) is much greater than in the perpendicular direction (y). Frequently, due to limitations in the number of independent electrodes in the multi-electrode system, the surveys use a roll-along system with a small number of parallel survey lines aligned along the x-direction. The `Compare R' array optimization method previously used for 2-D surveys is adapted for such 3-D surveys. Offset versions of the inline arrays used in 2-D surveys are included in the number of possible arrays (the comprehensive data set) to improve the sensitivity to structures in between the lines. The array geometric factor and its relative error are used to filter out potentially unstable arrays in the construction of the comprehensive data set. Comparisons of the conventional (consisting of dipole-dipole and Wenner-Schlumberger arrays) and optimized arrays are made using a synthetic model and experimental measurements in a tank. The tests show that structures located between the lines are better resolved with the optimized arrays. The optimized arrays also have significantly better depth resolution compared to the conventional arrays.

  7. 3D Imaging of Nanoparticle Distribution in Biological Tissue by Laser-Induced Breakdown Spectroscopy

    PubMed Central

    Gimenez, Y.; Busser, B.; Trichard, F.; Kulesza, A.; Laurent, J. M.; Zaun, V.; Lux, F.; Benoit, J. M.; Panczer, G.; Dugourd, P.; Tillement, O.; Pelascini, F.; Sancey, L.; Motto-Ros, V.

    2016-01-01

    Nanomaterials represent a rapidly expanding area of research with huge potential for future medical applications. Nanotechnology indeed promises to revolutionize diagnostics, drug delivery, gene therapy, and many other areas of research. For any biological investigation involving nanomaterials, it is crucial to study the behavior of such nano-objects within tissues to evaluate both their efficacy and their toxicity. Here, we provide the first account of 3D label-free nanoparticle imaging at the entire-organ scale. The technology used is known as laser-induced breakdown spectroscopy (LIBS) and possesses several advantages such as speed of operation, ease of use and full compatibility with optical microscopy. We then used two different but complementary approaches to achieve 3D elemental imaging with LIBS: a volume reconstruction of a sliced organ and in-depth analysis. This proof-of-concept study demonstrates the quantitative imaging of both endogenous and exogenous elements within entire organs and paves the way for innumerable applications. PMID:27435424

  8. Image informatics for studying signal transduction in cells interacting with 3D matrices

    NASA Astrophysics Data System (ADS)

    Tzeranis, Dimitrios S.; Guo, Jin; Chen, Chengpin; Yannas, Ioannis V.; Wei, Xunbin; So, Peter T. C.

    2014-03-01

    Cells sense and respond to chemical stimuli on their environment via signal transduction pathways, complex networks of proteins whose interactions transmit chemical information. This work describes an implementation of image informatics, imaging-based methodologies for studying signal transduction networks. The methodology developed focuses on studying signal transduction networks in cells that interact with 3D matrices. It utilizes shRNA-based knock down of network components, 3D high-content imaging of cells inside the matrix by spectral multi-photon microscopy, and single-cell quantification using features that describe both cell morphology and cell-matrix adhesion pattern. The methodology is applied in a pilot study of TGFβ signaling via the SMAD pathway in fibroblasts cultured inside porous collagen-GAG scaffolds, biomaterials similar to the ones used clinically to induce skin regeneration. Preliminary results suggest that knocking down all rSMAD components affects fibroblast response to TGFβ1 and TGFβ3 isoforms in different ways, and suggest a potential role for SMAD1 and SMAD5 in regulating TGFβ isoform response. These preliminary results need to be verified with proteomic results that can provide solid evidence about the particular role of individual components of the SMAD pathway.

  9. Pseudo-3D Imaging With The DICOM-8

    NASA Astrophysics Data System (ADS)

    Shalev, S.; Arenson, J.; Kettner, B.

    1985-09-01

    We have developed the DICOM.-8 digital imaging computer for video image acquisition, processing and display. It is a low-cost mobile systems based on a Z80 microcomputer which controls access to two 512 x 512 x 8-bit image planes through a real-time video arithmetic unit. Image presentation capabilities include orthographic images, isometric plots with hidden-line suppression, real-time mask subtraction, binocular red/green stereo, and volumetric imaging with both geometrical and density windows under operator interactive control. Examples are shown for multiplane series of CT images.

  10. High-content 3D multicolor super-resolution localization microscopy.

    PubMed

    Pereira, Pedro M; Almada, Pedro; Henriques, Ricardo

    2015-01-01

    Super-resolution (SR) methodologies permit the visualization of cellular structures at near-molecular scale (1-30 nm), enabling novel mechanistic analysis of key events in cell biology not resolvable by conventional fluorescence imaging (∼300-nm resolution). When this level of detail is combined with computing power and fast and reliable analysis software, high-content screenings using SR becomes a practical option to address multiple biological questions. The importance of combining these powerful analytical techniques cannot be ignored, as they can address phenotypic changes on the molecular scale and in a statistically robust manner. In this work, we suggest an easy-to-implement protocol that can be applied to set up a high-content 3D SR experiment with user-friendly and freely available software. The protocol can be divided into two main parts: chamber and sample preparation, where a protocol to set up a direct STORM (dSTORM) sample is presented; and a second part where a protocol for image acquisition and analysis is described. We intend to take the reader step-by-step through the experimental process highlighting possible experimental bottlenecks and possible improvements based on recent developments in the field.

  11. High-content 3D multicolor super-resolution localization microscopy.

    PubMed

    Pereira, Pedro M; Almada, Pedro; Henriques, Ricardo

    2015-01-01

    Super-resolution (SR) methodologies permit the visualization of cellular structures at near-molecular scale (1-30 nm), enabling novel mechanistic analysis of key events in cell biology not resolvable by conventional fluorescence imaging (∼300-nm resolution). When this level of detail is combined with computing power and fast and reliable analysis software, high-content screenings using SR becomes a practical option to address multiple biological questions. The importance of combining these powerful analytical techniques cannot be ignored, as they can address phenotypic changes on the molecular scale and in a statistically robust manner. In this work, we suggest an easy-to-implement protocol that can be applied to set up a high-content 3D SR experiment with user-friendly and freely available software. The protocol can be divided into two main parts: chamber and sample preparation, where a protocol to set up a direct STORM (dSTORM) sample is presented; and a second part where a protocol for image acquisition and analysis is described. We intend to take the reader step-by-step through the experimental process highlighting possible experimental bottlenecks and possible improvements based on recent developments in the field. PMID:25640426

  12. 3D Prostate Segmentation of Ultrasound Images Combining Longitudinal Image Registration and Machine Learning

    PubMed Central

    Yang, Xiaofeng; Fei, Baowei

    2012-01-01

    We developed a three-dimensional (3D) segmentation method for transrectal ultrasound (TRUS) images, which is based on longitudinal image registration and machine learning. Using longitudinal images of each individual patient, we register previously acquired images to the new images of the same subject. Three orthogonal Gabor filter banks were used to extract texture features from each registered image. Patient-specific Gabor features from the registered images are used to train kernel support vector machines (KSVMs) and then to segment the newly acquired prostate image. The segmentation method was tested in TRUS data from five patients. The average surface distance between our and manual segmentation is 1.18 ± 0.31 mm, indicating that our automatic segmentation method based on longitudinal image registration is feasible for segmenting the prostate in TRUS images. PMID:24027622

  13. 3D super-resolved in vitro multiphoton microscopy by saturation of excitation.

    PubMed

    Nguyen, Anh Dung; Duport, François; Bouwens, Arno; Vanholsbeeck, Frédérique; Egrise, Dominique; Van Simaeys, Gaetan; Emplit, Philippe; Goldman, Serge; Gorza, Simon-Pierre

    2015-08-24

    We demonstrate a significant resolution enhancement beyond the conventional limit in multiphoton microscopy (MPM) using saturated excitation of fluorescence. Our technique achieves super-resolved imaging by temporally modulating the excitation laser-intensity and demodulating the higher harmonics from the saturated fluorescence signal. The improvement of the lateral and axial resolutions is measured on a sample of fluorescent microspheres. While the third harmonic already provides an enhanced resolution, we show that a further improvement can be obtained with an appropriate linear combination of the demodulated harmonics. Finally, we present in vitro imaging of fluorescent microspheres incorporated in HeLa cells to show that this technique performs well in biological samples. PMID:26368235

  14. Increasing the depth of field in Multiview 3D images

    NASA Astrophysics Data System (ADS)

    Lee, Beom-Ryeol; Son, Jung-Young; Yano, Sumio; Jung, Ilkwon

    2016-06-01

    A super-multiview condition simulator which can project up to four different view images to each eye is introduced. This simulator with the image having both disparity and perspective informs that the depth of field (DOF) will be extended to more than the default DOF values as the number of simultaneously but separately projected different view images to each eye increase. The DOF range can be extended to near 2 diopters with the four simultaneous view images. However, the DOF value increments are not prominent as the image with both disparity and perspective with the image with disparity only.

  15. D3D augmented reality imaging system: proof of concept in mammography

    PubMed Central

    Douglas, David B; Petricoin, Emanuel F; Liotta, Lance; Wilson, Eugene

    2016-01-01

    Purpose The purpose of this article is to present images from simulated breast microcalcifications and assess the pattern of the microcalcifications with a technical development called “depth 3-dimensional (D3D) augmented reality”. Materials and methods A computer, head display unit, joystick, D3D augmented reality software, and an in-house script of simulated data of breast microcalcifications in a ductal distribution were used. No patient data was used and no statistical analysis was performed. Results The D3D augmented reality system demonstrated stereoscopic depth perception by presenting a unique image to each eye, focal point convergence, head position tracking, 3D cursor, and joystick fly-through. Conclusion The D3D augmented reality imaging system offers image viewing with depth perception and focal point convergence. The D3D augmented reality system should be tested to determine its utility in clinical practice. PMID:27563261

  16. Flash trajectory imaging of target 3D motion

    NASA Astrophysics Data System (ADS)

    Wang, Xinwei; Zhou, Yan; Fan, Songtao; He, Jun; Liu, Yuliang

    2011-03-01

    We present a flash trajectory imaging technique which can directly obtain target trajectory and realize non-contact measurement of motion parameters by range-gated imaging and time delay integration. Range-gated imaging gives the range of targets and realizes silhouette detection which can directly extract targets from complex background and decrease the complexity of moving target image processing. Time delay integration increases information of one single frame of image so that one can directly gain the moving trajectory. In this paper, we have studied the algorithm about flash trajectory imaging and performed initial experiments which successfully obtained the trajectory of a falling badminton. Our research demonstrates that flash trajectory imaging is an effective approach to imaging target trajectory and can give motion parameters of moving targets.

  17. [3D Super-resolution Reconstruction and Visualization of Pulmonary Nodules from CT Image].

    PubMed

    Wang, Bing; Fan, Xing; Yang, Ying; Tian, Xuedong; Gu, Lixu

    2015-08-01

    The aim of this study was to propose an algorithm for three-dimensional projection onto convex sets (3D POCS) to achieve super resolution reconstruction of 3D lung computer tomography (CT) images, and to introduce multi-resolution mixed display mode to make 3D visualization of pulmonary nodules. Firstly, we built the low resolution 3D images which have spatial displacement in sub pixel level between each other and generate the reference image. Then, we mapped the low resolution images into the high resolution reference image using 3D motion estimation and revised the reference image based on the consistency constraint convex sets to reconstruct the 3D high resolution images iteratively. Finally, we displayed the different resolution images simultaneously. We then estimated the performance of provided method on 5 image sets and compared them with those of 3 interpolation reconstruction methods. The experiments showed that the performance of 3D POCS algorithm was better than that of 3 interpolation reconstruction methods in two aspects, i.e., subjective and objective aspects, and mixed display mode is suitable to the 3D visualization of high resolution of pulmonary nodules.

  18. Advanced 2D-3D registration for endovascular aortic interventions: addressing dissimilarity in images

    NASA Astrophysics Data System (ADS)

    Demirci, Stefanie; Kutter, Oliver; Manstad-Hulaas, Frode; Bauernschmitt, Robert; Navab, Nassir

    2008-03-01

    In the current clinical workflow of minimally invasive aortic procedures navigation tasks are performed under 2D or 3D angiographic imaging. Many solutions for navigation enhancement suggest an integration of the preoperatively acquired computed tomography angiography (CTA) in order to provide the physician with more image information and reduce contrast injection and radiation exposure. This requires exact registration algorithms that align the CTA volume to the intraoperative 2D or 3D images. Additional to the real-time constraint, the registration accuracy should be independent of image dissimilarities due to varying presence of medical instruments and contrast agent. In this paper, we propose efficient solutions for image-based 2D-3D and 3D-3D registration that reduce the dissimilarities by image preprocessing, e.g. implicit detection and segmentation, and adaptive weights introduced into the registration procedure. Experiments and evaluations are conducted on real patient data.

  19. Analyzing Remodeling of Cardiac Tissue: A Comprehensive Approach Based on Confocal Microscopy and 3D Reconstructions.

    PubMed

    Seidel, Thomas; Edelmann, J-C; Sachse, Frank B

    2016-05-01

    Microstructural characterization of cardiac tissue and its remodeling in disease is a crucial step in many basic research projects. We present a comprehensive approach for three-dimensional characterization of cardiac tissue at the submicrometer scale. We developed a compression-free mounting method as well as labeling and imaging protocols that facilitate acquisition of three-dimensional image stacks with scanning confocal microscopy. We evaluated the approach with normal and infarcted ventricular tissue. We used the acquired image stacks for segmentation, quantitative analysis and visualization of important tissue components. In contrast to conventional mounting, compression-free mounting preserved cell shapes, capillary lumens and extracellular laminas. Furthermore, the new approach and imaging protocols resulted in high signal-to-noise ratios at depths up to 60 µm. This allowed extensive analyzes revealing major differences in volume fractions and distribution of cardiomyocytes, blood vessels, fibroblasts, myofibroblasts and extracellular space in control vs. infarct border zone. Our results show that the developed approach yields comprehensive data on microstructure of cardiac tissue and its remodeling in disease. In contrast to other approaches, it allows quantitative assessment of all major tissue components. Furthermore, we suggest that the approach will provide important data for physiological models of cardiac tissue at the submicrometer scale. PMID:26399990

  20. Analyzing Remodeling of Cardiac Tissue: A Comprehensive Approach Based on Confocal Microscopy and 3D Reconstructions.

    PubMed

    Seidel, Thomas; Edelmann, J-C; Sachse, Frank B

    2016-05-01

    Microstructural characterization of cardiac tissue and its remodeling in disease is a crucial step in many basic research projects. We present a comprehensive approach for three-dimensional characterization of cardiac tissue at the submicrometer scale. We developed a compression-free mounting method as well as labeling and imaging protocols that facilitate acquisition of three-dimensional image stacks with scanning confocal microscopy. We evaluated the approach with normal and infarcted ventricular tissue. We used the acquired image stacks for segmentation, quantitative analysis and visualization of important tissue components. In contrast to conventional mounting, compression-free mounting preserved cell shapes, capillary lumens and extracellular laminas. Furthermore, the new approach and imaging protocols resulted in high signal-to-noise ratios at depths up to 60 µm. This allowed extensive analyzes revealing major differences in volume fractions and distribution of cardiomyocytes, blood vessels, fibroblasts, myofibroblasts and extracellular space in control vs. infarct border zone. Our results show that the developed approach yields comprehensive data on microstructure of cardiac tissue and its remodeling in disease. In contrast to other approaches, it allows quantitative assessment of all major tissue components. Furthermore, we suggest that the approach will provide important data for physiological models of cardiac tissue at the submicrometer scale.

  1. Multiphoton Microscopy for Ophthalmic Imaging

    PubMed Central

    Gibson, Emily A.; Masihzadeh, Omid; Lei, Tim C.; Ammar, David A.; Kahook, Malik Y.

    2011-01-01

    We review multiphoton microscopy (MPM) including two-photon autofluorescence (2PAF), second harmonic generation (SHG), third harmonic generation (THG), fluorescence lifetime (FLIM), and coherent anti-Stokes Raman Scattering (CARS) with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the particular strength that each has for functional tissue imaging. MPM is compared with current clinical ophthalmological imaging techniques such as reflectance confocal microscopy, optical coherence tomography, and fluorescence imaging. In addition, we discuss the future prospects for MPM in disease detection and clinical monitoring of disease progression, understanding fundamental disease mechanisms, and real-time monitoring of drug delivery. PMID:21274261

  2. 3D image display of fetal ultrasonic images by thin shell

    NASA Astrophysics Data System (ADS)

    Wang, Shyh-Roei; Sun, Yung-Nien; Chang, Fong-Ming; Jiang, Ching-Fen

    1999-05-01

    Due to the properties of convenience and non-invasion, ultrasound has become an essential tool for diagnosis of fetal abnormality during women pregnancy in obstetrics. However, the 'noisy and blurry' nature of ultrasound data makes the rendering of the data a challenge in comparison with MRI and CT images. In spite of the speckle noise, the unwanted objects usually occlude the target to be observed. In this paper, we proposed a new system that can effectively depress the speckle noise, extract the target object, and clearly render the 3D fetal image in almost real-time from 3D ultrasound image data. The system is based on a deformable model that detects contours of the object according to the local image feature of ultrasound. Besides, in order to accelerate rendering speed, a thin shell is defined to separate the observed organ from unrelated structures depending on those detected contours. In this way, we can support quick 3D display of ultrasound, and the efficient visualization of 3D fetal ultrasound thus becomes possible.

  3. Infrared imaging of the polymer 3D-printing process

    NASA Astrophysics Data System (ADS)

    Dinwiddie, Ralph B.; Kunc, Vlastimil; Lindal, John M.; Post, Brian; Smith, Rachel J.; Love, Lonnie; Duty, Chad E.

    2014-05-01

    Both mid-wave and long-wave IR cameras are used to measure various temperature profiles in thermoplastic parts as they are printed. Two significantly different 3D-printers are used in this study. The first is a small scale commercially available Solidoodle 3 printer, which prints parts with layer thicknesses on the order of 125μm. The second printer used is a "Big Area Additive Manufacturing" (BAAM) 3D-printer developed at Oak Ridge National Laboratory. The BAAM prints parts with a layer thicknesses of 4.06 mm. Of particular interest is the temperature of the previously deposited layer as the new hot layer is about to be extruded onto it. The two layers are expected have a stronger bond if the temperature of the substrate layer is above the glass transition temperature. This paper describes the measurement technique and results for a study of temperature decay and substrate layer temperature for ABS thermoplastic with and without the addition of chopped carbon fibers.

  4. 3D breast image registration--a review.

    PubMed

    Sivaramakrishna, Radhika

    2005-02-01

    Image registration is an important problem in breast imaging. It is used in a wide variety of applications that include better visualization of lesions on pre- and post-contrast breast MRI images, speckle tracking and image compounding in breast ultrasound images, alignment of positron emission, and standard mammography images on hybrid machines et cetera. It is a prerequisite to align images taken at different times to isolate small interval lesions. Image registration also has useful applications in monitoring cancer therapy. The field of breast image registration has gained considerable interest in recent years. While the primary focus of interest continues to be the registration of pre- and post-contrast breast MRI images, other areas like breast ultrasound registration have gained more attention in recent years. The focus of registration algorithms has also shifted from control point based semi-automated techniques, to more sophisticated voxel based automated techniques that use mutual information as a similarity measure. This paper visits the problem of breast image registration and provides an overview of the current state-of-the-art in this area. PMID:15649086

  5. 3D inclusion trail geometry determination within individual porphyroblasts using reflected light optical microscopy of oriented blocks

    NASA Astrophysics Data System (ADS)

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

    2015-04-01

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

  6. Real-time computer-generated integral imaging and 3D image calibration for augmented reality surgical navigation.

    PubMed

    Wang, Junchen; Suenaga, Hideyuki; Liao, Hongen; Hoshi, Kazuto; Yang, Liangjing; Kobayashi, Etsuko; Sakuma, Ichiro

    2015-03-01

    Autostereoscopic 3D image overlay for augmented reality (AR) based surgical navigation has been studied and reported many times. For the purpose of surgical overlay, the 3D image is expected to have the same geometric shape as the original organ, and can be transformed to a specified location for image overlay. However, how to generate a 3D image with high geometric fidelity and quantitative evaluation of 3D image's geometric accuracy have not been addressed. This paper proposes a graphics processing unit (GPU) based computer-generated integral imaging pipeline for real-time autostereoscopic 3D display, and an automatic closed-loop 3D image calibration paradigm for displaying undistorted 3D images. Based on the proposed methods, a novel AR device for 3D image surgical overlay is presented, which mainly consists of a 3D display, an AR window, a stereo camera for 3D measurement, and a workstation for information processing. The evaluation on the 3D image rendering performance with 2560×1600 elemental image resolution shows the rendering speeds of 50-60 frames per second (fps) for surface models, and 5-8 fps for large medical volumes. The evaluation of the undistorted 3D image after the calibration yields sub-millimeter geometric accuracy. A phantom experiment simulating oral and maxillofacial surgery was also performed to evaluate the proposed AR overlay device in terms of the image registration accuracy, 3D image overlay accuracy, and the visual effects of the overlay. The experimental results show satisfactory image registration and image overlay accuracy, and confirm the system usability.

  7. 3-D Target Location from Stereoscopic SAR Images

    SciTech Connect

    DOERRY,ARMIN W.

    1999-10-01

    SAR range-Doppler images are inherently 2-dimensional. Targets with a height offset lay over onto offset range and azimuth locations. Just which image locations are laid upon depends on the imaging geometry, including depression angle, squint angle, and target bearing. This is the well known layover phenomenon. Images formed with different aperture geometries will exhibit different layover characteristics. These differences can be exploited to ascertain target height information, in a stereoscopic manner. Depending on the imaging geometries, height accuracy can be on the order of horizontal position accuracies, thereby rivaling the best IFSAR capabilities in fine resolution SAR images. All that is required for this to work are two distinct passes with suitably different geometries from any plain old SAR.

  8. A survey of clearing techniques for 3D imaging of tissues with special reference to connective tissue.

    PubMed

    Azaripour, Adriano; Lagerweij, Tonny; Scharfbillig, Christina; Jadczak, Anna Elisabeth; Willershausen, Brita; Van Noorden, Cornelis J F

    2016-08-01

    For 3-dimensional (3D) imaging of a tissue, 3 methodological steps are essential and their successful application depends on specific characteristics of the type of tissue. The steps are 1° clearing of the opaque tissue to render it transparent for microscopy, 2° fluorescence labeling of the tissues and 3° 3D imaging. In the past decades, new methodologies were introduced for the clearing steps with their specific advantages and disadvantages. Most clearing techniques have been applied to the central nervous system and other organs that contain relatively low amounts of connective tissue including extracellular matrix. However, tissues that contain large amounts of extracellular matrix such as dermis in skin or gingiva are difficult to clear. The present survey lists methodologies that are available for clearing of tissues for 3D imaging. We report here that the BABB method using a mixture of benzyl alcohol and benzyl benzoate and iDISCO using dibenzylether (DBE) are the most successful methods for clearing connective tissue-rich gingiva and dermis of skin for 3D histochemistry and imaging of fluorescence using light-sheet microscopy.

  9. Thin client performance for remote 3-D image display.

    PubMed

    Lai, Albert; Nieh, Jason; Laine, Andrew; Starren, Justin

    2003-01-01

    Several trends in biomedical computing are converging in a way that will require new approaches to telehealth image display. Image viewing is becoming an "anytime, anywhere" activity. In addition, organizations are beginning to recognize that healthcare providers are highly mobile and optimal care requires providing information wherever the provider and patient are. Thin-client computing is one way to support image viewing this complex environment. However little is known about the behavior of thin client systems in supporting image transfer in modern heterogeneous networks. Our results show that using thin-clients can deliver acceptable performance over conditions commonly seen in wireless networks if newer protocols optimized for these conditions are used.

  10. Pragmatic fully 3D image reconstruction for the MiCES mouse imaging PET scanner

    NASA Astrophysics Data System (ADS)

    Lee, Kisung; Kinahan, Paul E.; Fessler, Jeffrey A.; Miyaoka, Robert S.; Janes, Marie; Lewellen, Tom K.

    2004-10-01

    We present a pragmatic approach to image reconstruction for data from the micro crystal elements system (MiCES) fully 3D mouse imaging positron emission tomography (PET) scanner under construction at the University of Washington. Our approach is modelled on fully 3D image reconstruction used in clinical PET scanners, which is based on Fourier rebinning (FORE) followed by 2D iterative image reconstruction using ordered-subsets expectation-maximization (OSEM). The use of iterative methods allows modelling of physical effects (e.g., statistical noise, detector blurring, attenuation, etc), while FORE accelerates the reconstruction process by reducing the fully 3D data to a stacked set of independent 2D sinograms. Previous investigations have indicated that non-stationary detector point-spread response effects, which are typically ignored for clinical imaging, significantly impact image quality for the MiCES scanner geometry. To model the effect of non-stationary detector blurring (DB) in the FORE+OSEM(DB) algorithm, we have added a factorized system matrix to the ASPIRE reconstruction library. Initial results indicate that the proposed approach produces an improvement in resolution without an undue increase in noise and without a significant increase in the computational burden. The impact on task performance, however, remains to be evaluated.

  11. 3-D ultrafast Doppler imaging applied to the noninvasive mapping of blood vessels in vivo.

    PubMed

    Provost, Jean; Papadacci, Clement; Demene, Charlie; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2015-08-01

    Ultrafast Doppler imaging was introduced as a technique to quantify blood flow in an entire 2-D field of view, expanding the field of application of ultrasound imaging to the highly sensitive anatomical and functional mapping of blood vessels. We have recently developed 3-D ultrafast ultrasound imaging, a technique that can produce thousands of ultrasound volumes per second, based on a 3-D plane and diverging wave emissions, and demonstrated its clinical feasibility in human subjects in vivo. In this study, we show that noninvasive 3-D ultrafast power Doppler, pulsed Doppler, and color Doppler imaging can be used to perform imaging of blood vessels in humans when using coherent compounding of 3-D tilted plane waves. A customized, programmable, 1024-channel ultrasound system was designed to perform 3-D ultrafast imaging. Using a 32 × 32, 3-MHz matrix phased array (Vermon, Tours, France), volumes were beamformed by coherently compounding successive tilted plane wave emissions. Doppler processing was then applied in a voxel-wise fashion. The proof of principle of 3-D ultrafast power Doppler imaging was first performed by imaging Tygon tubes of various diameters, and in vivo feasibility was demonstrated by imaging small vessels in the human thyroid. Simultaneous 3-D color and pulsed Doppler imaging using compounded emissions were also applied in the carotid artery and the jugular vein in one healthy volunteer.

  12. 3-D ultrafast Doppler imaging applied to the noninvasive mapping of blood vessels in vivo.

    PubMed

    Provost, Jean; Papadacci, Clement; Demene, Charlie; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2015-08-01

    Ultrafast Doppler imaging was introduced as a technique to quantify blood flow in an entire 2-D field of view, expanding the field of application of ultrasound imaging to the highly sensitive anatomical and functional mapping of blood vessels. We have recently developed 3-D ultrafast ultrasound imaging, a technique that can produce thousands of ultrasound volumes per second, based on a 3-D plane and diverging wave emissions, and demonstrated its clinical feasibility in human subjects in vivo. In this study, we show that noninvasive 3-D ultrafast power Doppler, pulsed Doppler, and color Doppler imaging can be used to perform imaging of blood vessels in humans when using coherent compounding of 3-D tilted plane waves. A customized, programmable, 1024-channel ultrasound system was designed to perform 3-D ultrafast imaging. Using a 32 × 32, 3-MHz matrix phased array (Vermon, Tours, France), volumes were beamformed by coherently compounding successive tilted plane wave emissions. Doppler processing was then applied in a voxel-wise fashion. The proof of principle of 3-D ultrafast power Doppler imaging was first performed by imaging Tygon tubes of various diameters, and in vivo feasibility was demonstrated by imaging small vessels in the human thyroid. Simultaneous 3-D color and pulsed Doppler imaging using compounded emissions were also applied in the carotid artery and the jugular vein in one healthy volunteer. PMID:26276956

  13. Review of three-dimensional (3D) surface imaging for oncoplastic, reconstructive and aesthetic breast surgery.

    PubMed

    O'Connell, Rachel L; Stevens, Roger J G; Harris, Paul A; Rusby, Jennifer E

    2015-08-01

    Three-dimensional surface imaging (3D-SI) is being marketed as a tool in aesthetic breast surgery. It has recently also been studied in the objective evaluation of cosmetic outcome of oncological procedures. The aim of this review is to summarise the use of 3D-SI in oncoplastic, reconstructive and aesthetic breast surgery. An extensive literature review was undertaken to identify published studies. Two reviewers independently screened all abstracts and selected relevant articles using specific inclusion criteria. Seventy two articles relating to 3D-SI for breast surgery were identified. These covered endpoints such as image acquisition, calculations and data obtainable, comparison of 3D and 2D imaging and clinical research applications of 3D-SI. The literature provides a favourable view of 3D-SI. However, evidence of its superiority over current methods of clinical decision making, surgical planning, communication and evaluation of outcome is required before it can be accepted into mainstream practice.

  14. Computation of tooth axes of existent and missing teeth from 3D CT images.

    PubMed

    Wang, Yang; Wu, Lin; Guo, Huayan; Qiu, Tiantian; Huang, Yuanliang; Lin, Bin; Wang, Lisheng

    2015-12-01

    Orientations of tooth axes are important quantitative information used in dental diagnosis and surgery planning. However, their computation is a complex problem, and the existing methods have respective limitations. This paper proposes new methods to compute 3D tooth axes from 3D CT images for existent teeth with single root or multiple roots and to estimate 3D tooth axes from 3D CT images for missing teeth. The tooth axis of a single-root tooth will be determined by segmenting the pulp cavity of the tooth and computing the principal direction of the pulp cavity, and the estimation of tooth axes of the missing teeth is modeled as an interpolation problem of some quaternions along a 3D curve. The proposed methods can either avoid the difficult teeth segmentation problem or improve the limitations of existing methods. Their effectiveness and practicality are demonstrated by experimental results of different 3D CT images from the clinic.

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

    NASA Astrophysics Data System (ADS)

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

    2013-02-01

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

  16. 3-D visualization and identification of biological microorganisms using partially temporal incoherent light in-line computational holographic imaging.

    PubMed

    Moon, Inkyu; Javidi, Bahram

    2008-12-01

    We present a new method for three-dimensional (3-D) visualization and identification of biological microorganisms using partially temporal incoherent light in-line (PTILI) computational holographic imaging and multivariate statistical methods. For 3-D data acquisition of biological microorganisms, the band-pass filtered white light is used to illuminate a biological sample. The transversely and longitudinally diffracted pattern of the biological sample is magnified by microscope objective (MO) and is optically recorded with an image sensor array interfaced with a computer. Three-dimensional reconstruction of the biological sample from the diffraction pattern is accomplished by using computational Fresnel propagation method. Principal components analysis and nonparametric inference algorithms are applied to the 3-D complex amplitude biological sample for identification purposes. Experiments indicate that the proposed system can be useful for identifying biological microorganisms. To the best of our knowledge, this is the first report on using PTILI computational holographic microscopy for identification of biological microorganisms.

  17. Focussed ion beam serial sectioning and imaging of monolithic materials for 3D reconstruction and morphological parameter evaluation.

    PubMed

    Vázquez, Mercedes; Moore, David; He, Xiaoyun; Ben Azouz, Aymen; Nesterenko, Ekaterina; Nesterenko, Pavel; Paull, Brett; Brabazon, Dermot

    2014-01-01

    A new characterisation method, based on the utilisation of focussed ion beam-scanning electron microscopy (FIB-SEM), has been employed for the evaluation of morphological parameters in porous monolithic materials. Sample FIB serial sectioning, SEM imaging and image processing techniques were used to extract the pore boundaries and reconstruct the 3D porous structure of carbon and silica-based monoliths. Since silica is a non-conducting material, a commercial silica monolith modified with activated carbon was employed instead to minimise the charge build-up during FIB sectioning. This work therefore presents a novel methodology that can be successfully employed for 3D reconstruction of porous monolithic materials which are or can be made conductive through surface or bulk modification. Furthermore, the 3D reconstructions were used for calculation of the monolith macroporosity, which was in good agreement with the porosity values obtained by mercury intrusion porosimetry (MIP).

  18. Quality assessment of stereoscopic 3D image compression by binocular integration behaviors.

    PubMed

    Lin, Yu-Hsun; Wu, Ja-Ling

    2014-04-01

    The objective approaches of 3D image quality assessment play a key role for the development of compression standards and various 3D multimedia applications. The quality assessment of 3D images faces more new challenges, such as asymmetric stereo compression, depth perception, and virtual view synthesis, than its 2D counterparts. In addition, the widely used 2D image quality metrics (e.g., PSNR and SSIM) cannot be directly applied to deal with these newly introduced challenges. This statement can be verified by the low correlation between the computed objective measures and the subjectively measured mean opinion scores (MOSs), when 3D images are the tested targets. In order to meet these newly introduced challenges, in this paper, besides traditional 2D image metrics, the binocular integration behaviors-the binocular combination and the binocular frequency integration, are utilized as the bases for measuring the quality of stereoscopic 3D images. The effectiveness of the proposed metrics is verified by conducting subjective evaluations on publicly available stereoscopic image databases. Experimental results show that significant consistency could be reached between the measured MOS and the proposed metrics, in which the correlation coefficient between them can go up to 0.88. Furthermore, we found that the proposed metrics can also address the quality assessment of the synthesized color-plus-depth 3D images well. Therefore, it is our belief that the binocular integration behaviors are important factors in the development of objective quality assessment for 3D images.

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

    PubMed

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

    2012-07-01

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

  20. Estimation of the degree of polarization in low-light 3D integral imaging

    NASA Astrophysics Data System (ADS)

    Carnicer, Artur; Javidi, Bahram

    2016-06-01

    The calculation of the Stokes Parameters and the Degree of Polarization in 3D integral images requires a careful manipulation of the polarimetric elemental images. This fact is particularly important if the scenes are taken in low-light conditions. In this paper, we show that the Degree of Polarization can be effectively estimated even when elemental images are recorded with few photons. The original idea was communicated in [A. Carnicer and B. Javidi, "Polarimetric 3D integral imaging in photon-starved conditions," Opt. Express 23, 6408-6417 (2015)]. First, we use the Maximum Likelihood Estimation approach for generating the 3D integral image. Nevertheless, this method produces very noisy images and thus, the degree of polarization cannot be calculated. We suggest using a Total Variation Denoising filter as a way to improve the quality of the generated 3D images. As a result, noise is suppressed but high frequency information is preserved. Finally, the degree of polarization is obtained successfully.

  1. Advances in Image Pre-Processing to Improve Automated 3d Reconstruction

    NASA Astrophysics Data System (ADS)

    Ballabeni, A.; Apollonio, F. I.; Gaiani, M.; Remondino, F.

    2015-02-01

    Tools and algorithms for automated image processing and 3D reconstruction purposes have become more and more available, giving the possibility to process any dataset of unoriented and markerless images. Typically, dense 3D point clouds (or texture 3D polygonal models) are produced at reasonable processing time. In this paper, we evaluate how the radiometric pre-processing of image datasets (particularly in RAW format) can help in improving the performances of state-of-the-art automated image processing tools. Beside a review of common pre-processing methods, an efficient pipeline based on color enhancement, image denoising, RGB to Gray conversion and image content enrichment is presented. The performed tests, partly reported for sake of space, demonstrate how an effective image pre-processing, which considers the entire dataset in analysis, can improve the automated orientation procedure and dense 3D point cloud reconstruction, even in case of poor texture scenarios.

  2. Computer-generated hologram for 3D scene from multi-view images

    NASA Astrophysics Data System (ADS)

    Chang, Eun-Young; Kang, Yun-Suk; Moon, KyungAe; Ho, Yo-Sung; Kim, Jinwoong

    2013-05-01

    Recently, the computer generated hologram (CGH) calculated from real existing objects is more actively investigated to support holographic video and TV applications. In this paper, we propose a method of generating a hologram of the natural 3-D scene from multi-view images in order to provide motion parallax viewing with a suitable navigation range. After a unified 3-D point source set describing the captured 3-D scene is obtained from multi-view images, a hologram pattern supporting motion-parallax is calculated from the set using a point-based CGH method. We confirmed that 3-D scenes are faithfully reconstructed using numerical reconstruction.

  3. ICER-3D: A Progressive Wavelet-Based Compressor for Hyperspectral Images

    NASA Technical Reports Server (NTRS)

    Kiely, A.; Klimesh, M.; Xie, H.; Aranki, N.

    2005-01-01

    ICER-3D is a progressive, wavelet-based compressor for hyperspectral images. ICER-3D is derived from the ICER image compressor. ICER-3D can provide lossless and lossy compression, and incorporates an error-containment scheme to limit the effects of data loss during transmission. The three-dime