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Sample records for 3d microscopy images

  1. 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.

  2. 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

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

    PubMed

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

    2005-01-01

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

  4. 3D Image Analysis of Geomaterials using Confocal Microscopy

    NASA Astrophysics Data System (ADS)

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

    2009-05-01

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

  5. 3D super-resolution imaging by localization microscopy.

    PubMed

    Magenau, Astrid; Gaus, Katharina

    2015-01-01

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

  6. 3D fluorescence anisotropy imaging using selective plane illumination microscopy

    PubMed Central

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

    2015-01-01

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

  7. 3D geometry-based quantification of colocalizations in multichannel 3D microscopy images of human soft tissue tumors.

    PubMed

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

    2010-08-01

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

  8. Cytology 3D structure formation based on optical microscopy images

    NASA Astrophysics Data System (ADS)

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

    2017-01-01

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

  9. Registration and 3D visualization of large microscopy images

    NASA Astrophysics Data System (ADS)

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

    2006-03-01

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

  10. 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

  11. 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.

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

    Duan, Yubo; Chen, Nanguang

    2015-11-15

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

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

    PubMed

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

    2010-06-22

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

  15. 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.

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

    PubMed

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

    2013-07-01

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

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

    PubMed

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

    2017-03-08

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

  18. 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.

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

    PubMed

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

    2010-07-01

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

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

    PubMed

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

    2017-02-10

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

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

    PubMed Central

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

    2017-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2017-02-01

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

  3. 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

  4. 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

  5. 3D structured illumination microscopy

    NASA Astrophysics Data System (ADS)

    Dougherty, William M.; Goodwin, Paul C.

    2011-03-01

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

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

    PubMed

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

    2013-01-01

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

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

    PubMed

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

    1996-11-01

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

  8. 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

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

    NASA Astrophysics Data System (ADS)

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

    2012-02-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-01-01

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

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

    SciTech Connect

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

    2016-01-15

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

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

    PubMed

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

    2009-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

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

  14. 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.

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

    PubMed

    Welsher, Kevin; Yang, Haw

    2015-01-01

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

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

    PubMed Central

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

    2014-01-01

    Despite the notable progress that has been made with nano-bio imaging probes, quantitative nanoscale imaging of multistructured specimens such as mammalian cells remains challenging due to their inherent structural complexity. Here, we successfully performed three-dimensional (3D) imaging of mammalian nuclei by combining coherent x-ray diffraction microscopy, explicitly visualizing nuclear substructures at several tens of nanometer resolution, and optical fluorescence microscopy, cross confirming the substructures with immunostaining. This demonstrates the successful application of coherent x-rays to obtain the 3D ultrastructure of mammalian nuclei and establishes a solid route to nanoscale imaging of complex specimens. PMID:25185543

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

    NASA Astrophysics Data System (ADS)

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

    2008-03-01

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

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

    PubMed

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

    2014-09-29

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

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

    Schonbrun, Ethan; Di Caprio, Giuseppe

    2016-10-01

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

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

    PubMed

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

    2015-03-24

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

  2. 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)

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

    PubMed

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

    2005-06-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

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

  5. 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.

  6. 3D myofibril imaging in live cardiomyocytes via hybrid SHG-TPEF microscopy

    NASA Astrophysics Data System (ADS)

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

    2011-03-01

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

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

    PubMed

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

    2010-03-01

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

  8. Atomic resolution 3D electron diffraction microscopy

    SciTech Connect

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

    2002-03-01

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

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

    PubMed Central

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

    2011-01-01

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

  10. 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

  11. 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.

  12. 3D photoacoustic imaging

    NASA Astrophysics Data System (ADS)

    Carson, Jeffrey J. L.; Roumeliotis, Michael; Chaudhary, Govind; Stodilka, Robert Z.; Anastasio, Mark A.

    2010-06-01

    Our group has concentrated on development of a 3D photoacoustic imaging system for biomedical imaging research. The technology employs a sparse parallel detection scheme and specialized reconstruction software to obtain 3D optical images using a single laser pulse. With the technology we have been able to capture 3D movies of translating point targets and rotating line targets. The current limitation of our 3D photoacoustic imaging approach is its inability ability to reconstruct complex objects in the field of view. This is primarily due to the relatively small number of projections used to reconstruct objects. However, in many photoacoustic imaging situations, only a few objects may be present in the field of view and these objects may have very high contrast compared to background. That is, the objects have sparse properties. Therefore, our work had two objectives: (i) to utilize mathematical tools to evaluate 3D photoacoustic imaging performance, and (ii) to test image reconstruction algorithms that prefer sparseness in the reconstructed images. Our approach was to utilize singular value decomposition techniques to study the imaging operator of the system and evaluate the complexity of objects that could potentially be reconstructed. We also compared the performance of two image reconstruction algorithms (algebraic reconstruction and l1-norm techniques) at reconstructing objects of increasing sparseness. We observed that for a 15-element detection scheme, the number of measureable singular vectors representative of the imaging operator was consistent with the demonstrated ability to reconstruct point and line targets in the field of view. We also observed that the l1-norm reconstruction technique, which is known to prefer sparseness in reconstructed images, was superior to the algebraic reconstruction technique. Based on these findings, we concluded (i) that singular value decomposition of the imaging operator provides valuable insight into the capabilities of

  13. 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

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

    PubMed

    Li, Qing; Deng, Zhigang

    2012-03-01

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

  15. 3D image reconstruction using optical sectioning in confocal scanning microscopy

    NASA Astrophysics Data System (ADS)

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

    2001-10-01

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

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

    PubMed Central

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

    2009-01-01

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

  17. 3D imaging of lung tissue by confocal microscopy and micro-CT

    NASA Astrophysics Data System (ADS)

    Kriete, Andres; Breithecker, Andreas; Rau, Wigbert D.

    2001-07-01

    Two complementary techniques for the imaging of tissue subunits are discussed. A computer guided light microscopic imaging technique is described first, which confocally resolves thick serial sections axially. The lateral area of interest is increased by scanning a mosaic of images in each plane. Subsequently, all images are fused digitally to form a highly resolved volume exhibiting the fine structure of complete respiratory units of lung. A different technique described is based on microtomography. This method allows to image volumes up to 3x3x3 cm at a resolution of up to 7 microns. Due to the lack of strong density differences, a contrast enhancement procedure is introduced which makes this technique applicable for the imaging of lung tissue. Imaging, visualization and analysis described here are parts of an ongoing project to model structure and to simulate function of tissue subunits and complete organs.

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

    PubMed

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

    2015-01-01

    Mycobacteria pose a threat to the world health today, with pathogenic and opportunistic bacteria causing tuberculosis and non-tuberculous disease in large parts of the population. Much is still unknown about the interplay between bacteria and host during infection and disease, and more research is needed to meet the challenge of drug resistance and inefficient vaccines. This work establishes a reliable and reproducible method for performing correlative imaging of human macrophages infected with mycobacteria at an ultra-high resolution and in 3D. Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) tomography is applied, together with confocal fluorescence microscopy for localization of appropriately infected cells. The method is based on an Aclar poly(chloro-tri-fluoro)ethylene substrate, micropatterned into an advantageous geometry by a simple thermomoulding process. The platform increases the throughput and quality of FIB/SEM tomography analyses, and was successfully applied to detail the intracellular environment of a whole mycobacterium-infected macrophage in 3D.

  19. 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

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

    PubMed

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

    2017-03-07

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-11-01

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

  2. 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

  3. 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.

  4. 3D microscopy - new powerful tools in geomaterials characterization

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

  5. 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

  6. Simple buffers for 3D STORM microscopy.

    PubMed

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

    2013-06-01

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

  7. Analysis of nuclear organization with TANGO, software for high-throughput quantitative analysis of 3D fluorescence microscopy images.

    PubMed

    Ollion, Jean; Cochennec, Julien; Loll, François; Escudé, Christophe; Boudier, Thomas

    2015-01-01

    The cell nucleus is a highly organized cellular organelle that contains the genome. An important step to understand the relationships between genome positioning and genome functions is to extract quantitative data from three-dimensional (3D) fluorescence imaging. However, such approaches are limited by the requirement for processing and analyzing large sets of images. Here we present a practical approach using TANGO (Tools for Analysis of Nuclear Genome Organization), an image analysis tool dedicated to the study of nuclear architecture. TANGO is a generic tool able to process large sets of images, allowing quantitative study of nuclear organization. In this chapter a practical description of the software is drawn in order to give an overview of its different concepts and functionalities. This description is illustrated with a precise example that can be performed step-by-step on experimental data provided on the website http://biophysique.mnhn.fr/tango/HomePage.

  8. 3D Viscoelastic traction force microscopy.

    PubMed

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

    2014-10-28

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

  9. 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.

  10. 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

  11. 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.

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

    PubMed Central

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

    2015-01-01

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

  13. 3D imaging of cement-based materials at submicron resolution by combining laser scanning confocal microscopy with serial sectioning.

    PubMed

    Yio, M H N; Mac, M J; Wong, H S; Buenfeld, N R

    2015-05-01

    In this paper, we present a new method to reconstruct large volumes of nontransparent porous materials at submicron resolution. The proposed method combines fluorescence laser scanning confocal microscopy with serial sectioning to produce a series of overlapping confocal z-stacks, which are then aligned and stitched based on phase correlation. The method can be extended in the XY plane to further increase the overall image volume. Resolution of the reconstructed image volume does not degrade with increase in sample size. We have used the method to image cementitious materials, hardened cement paste and concrete and the results obtained show that the method is reliable. Possible applications of the method such as three-dimensional characterization of the pores and microcracks in hardened concrete, three-dimensional particle shape characterization of cementitious materials and three-dimensional characterization of other porous materials such as rocks and bioceramics are discussed.

  14. 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.

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

    PubMed

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

    2015-11-01

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

  16. 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

  17. 3D differential phase contrast microscopy

    PubMed Central

    Chen, Michael; Tian, Lei; Waller, Laura

    2016-01-01

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

  18. Stabilizing 3D in vivo intravital microscopy images with an iteratively refined soft-tissue model for immunology experiments.

    PubMed

    Gómez-Conde, Iván; Caetano, Susana S; Tadokoro, Carlos E; Olivieri, David N

    2015-09-01

    We describe a set of new algorithms and a software tool, StabiTissue, for stabilizing in vivo intravital microscopy images that suffer from soft-tissue background movement. Because these images lack predetermined anchors and are dominated by noise, we use a pixel weighted image alignment together with a correction for nonlinear tissue deformations. We call this correction a poor man׳s diffeomorphic map since it ascertains the nonlinear regions of the image without resorting to a full integral equation method. To determine the quality of the image stabilization, we developed an ensemble sampling method that quantifies the coincidence between image pairs from randomly distributed image regions. We obtain global stabilization alignment through an iterative constrained simulated annealing optimization procedure. To show the accuracy of our algorithm with existing software, we measured the misalignment error rate in datasets taken from two different organs and compared the results to a similar and popular open-source solution. Present open-source stabilization software tools perform poorly because they do not treat the specific needs of the IV-2pM datasets with soft-tissue deformation, speckle noise, full 5D inter- and intra-stack motion error correction, and undefined anchors. In contrast, the results of our tests demonstrate that our method is more immune to noise and provides better performance for datasets' possessing nonlinear tissue deformations. As a practical application of our software, we show how our stabilization improves cell tracking, where the presence of background movement would degrade track information. We also provide a qualitative comparison of our software with other open-source libraries/applications. Our software is freely available at the open source repository http://sourceforge.net/projects/stabitissue/.

  19. 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

  20. 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.

  1. Active segmentation of 3D axonal images.

    PubMed

    Muralidhar, Gautam S; Gopinath, Ajay; Bovik, Alan C; Ben-Yakar, Adela

    2012-01-01

    We present an active contour framework for segmenting neuronal axons on 3D confocal microscopy data. Our work is motivated by the need to conduct high throughput experiments involving microfluidic devices and femtosecond lasers to study the genetic mechanisms behind nerve regeneration and repair. While most of the applications for active contours have focused on segmenting closed regions in 2D medical and natural images, there haven't been many applications that have focused on segmenting open-ended curvilinear structures in 2D or higher dimensions. The active contour framework we present here ties together a well known 2D active contour model [5] along with the physics of projection imaging geometry to yield a segmented axon in 3D. Qualitative results illustrate the promise of our approach for segmenting neruonal axons on 3D confocal microscopy data.

  2. 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.

  3. From Dynamic Live Cell Imaging to 3D Ultrastructure: Novel Integrated Methods for High Pressure Freezing and Correlative Light-Electron Microscopy

    PubMed Central

    Spiegelhalter, Coralie; Tosch, Valérie; Hentsch, Didier; Koch, Marc; Kessler, Pascal; Schwab, Yannick; Laporte, Jocelyn

    2010-01-01

    Background In cell biology, the study of proteins and organelles requires the combination of different imaging approaches, from live recordings with light microscopy (LM) to electron microscopy (EM). Methodology To correlate dynamic events in adherent cells with both ultrastructural and 3D information, we developed a method for cultured cells that combines confocal time-lapse images of GFP-tagged proteins with electron microscopy. With laser micro-patterned culture substrate, we created coordinates that were conserved at every step of the sample preparation and visualization processes. Specifically designed for cryo-fixation, this method allowed a fast freezing of dynamic events within seconds and their ultrastructural characterization. We provide examples of the dynamic oligomerization of GFP-tagged myotubularin (MTM1) phosphoinositides phosphatase induced by osmotic stress, and of the ultrastructure of membrane tubules dependent on amphiphysin 2 (BIN1) expression. Conclusion Accessible and versatile, we show that this approach is efficient to routinely correlate functional and dynamic LM with high resolution morphology by EM, with immuno-EM labeling, with 3D reconstruction using serial immuno-EM or tomography, and with scanning-EM. PMID:20140253

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

    PubMed

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

    2016-10-05

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

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

    PubMed

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

    2017-01-01

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

  6. Resolution in 3D in multifocal plane microscopy

    NASA Astrophysics Data System (ADS)

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

    2008-02-01

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

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

    PubMed

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

    2017-05-01

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

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

    PubMed

    Ezugwu, Sabastine; Ye, Hanyang; Fanchini, Giovanni

    2015-01-07

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

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

    NASA Astrophysics Data System (ADS)

    Ezugwu, Sabastine; Ye, Hanyang; Fanchini, Giovanni

    2014-11-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-08-01

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

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

    NASA Astrophysics Data System (ADS)

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

    1992-06-01

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

  12. Phase mask optimization for 3D parallax EDF microscopy

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

  13. True 3d Images and Their Applications

    NASA Astrophysics Data System (ADS)

    Wang, Z.; wang@hzgeospace., zheng.

    2012-07-01

    A true 3D image is a geo-referenced image. Besides having its radiometric information, it also has true 3Dground coordinates XYZ for every pixels of it. For a true 3D image, especially a true 3D oblique image, it has true 3D coordinates not only for building roofs and/or open grounds, but also for all other visible objects on the ground, such as visible building walls/windows and even trees. The true 3D image breaks the 2D barrier of the traditional orthophotos by introducing the third dimension (elevation) into the image. From a true 3D image, for example, people will not only be able to read a building's location (XY), but also its height (Z). true 3D images will fundamentally change, if not revolutionize, the way people display, look, extract, use, and represent the geospatial information from imagery. In many areas, true 3D images can make profound impacts on the ways of how geospatial information is represented, how true 3D ground modeling is performed, and how the real world scenes are presented. This paper first gives a definition and description of a true 3D image and followed by a brief review of what key advancements of geospatial technologies have made the creation of true 3D images possible. Next, the paper introduces what a true 3D image is made of. Then, the paper discusses some possible contributions and impacts the true 3D images can make to geospatial information fields. At the end, the paper presents a list of the benefits of having and using true 3D images and the applications of true 3D images in a couple of 3D city modeling projects.

  14. 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

  15. 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.

  16. 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.

  17. 3D carotid plaque MR Imaging

    PubMed Central

    Parker, Dennis L.

    2015-01-01

    SYNOPSIS There has been significant progress made in 3D carotid plaque magnetic resonance imaging techniques in recent years. 3D plaque imaging clearly represents the future in clinical use. With effective flow suppression techniques, choices of different contrast weighting acquisitions, and time-efficient imaging approaches, 3D plaque imaging offers flexible imaging plane and view angle analysis, large coverage, multi-vascular beds capability, and even can be used in fast screening. PMID:26610656

  18. 3D scanning Hall probe microscopy with 700 nm resolution

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

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

  20. 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

  1. Digital holography and 3-D imaging.

    PubMed

    Banerjee, Partha; Barbastathis, George; Kim, Myung; Kukhtarev, Nickolai

    2011-03-01

    This feature issue on Digital Holography and 3-D Imaging comprises 15 papers on digital holographic techniques and applications, computer-generated holography and encryption techniques, and 3-D display. It is hoped that future work in the area leads to innovative applications of digital holography and 3-D imaging to biology and sensing, and to the development of novel nonlinear dynamic digital holographic techniques.

  2. 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

    2014-10-07

    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.

  3. 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.

  4. 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.

  5. 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.

  6. 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

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

    PubMed

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

    2013-05-03

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

  8. 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.

  9. 3D high resolution pure optical photoacoustic microscopy

    NASA Astrophysics Data System (ADS)

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

    2012-02-01

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

  10. 3D imaging in forensic odontology.

    PubMed

    Evans, Sam; Jones, Carl; Plassmann, Peter

    2010-06-16

    This paper describes the investigation of a new 3D capture method for acquiring and subsequent forensic analysis of bite mark injuries on human skin. When documenting bite marks with standard 2D cameras errors in photographic technique can occur if best practice is not followed. Subsequent forensic analysis of the mark is problematic when a 3D structure is recorded into a 2D space. Although strict guidelines (BAFO) exist, these are time-consuming to follow and, due to their complexity, may produce errors. A 3D image capture and processing system might avoid the problems resulting from the 2D reduction process, simplifying the guidelines and reducing errors. Proposed Solution: a series of experiments are described in this paper to demonstrate that the potential of a 3D system might produce suitable results. The experiments tested precision and accuracy of the traditional 2D and 3D methods. A 3D image capture device minimises the amount of angular distortion, therefore such a system has the potential to create more robust forensic evidence for use in courts. A first set of experiments tested and demonstrated which method of forensic analysis creates the least amount of intra-operator error. A second set tested and demonstrated which method of image capture creates the least amount of inter-operator error and visual distortion. In a third set the effects of angular distortion on 2D and 3D methods of image capture were evaluated.

  11. 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.

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

    PubMed

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

    2015-05-01

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

  13. 3D integral imaging with optical processing

    NASA Astrophysics Data System (ADS)

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

    2008-04-01

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

  14. Adaptive-weighted cubic B-spline using lookup tables for fast and efficient axial resampling of 3D confocal microscopy images.

    PubMed

    Indhumathi, C; Cai, Y Y; Guan, Y Q; Opas, M; Zheng, J

    2012-01-01

    Confocal laser scanning microscopy has become a most powerful tool to visualize and analyze the dynamic behavior of cellular molecules. Photobleaching of fluorochromes is a major problem with confocal image acquisition that will lead to intensity attenuation. Photobleaching effect can be reduced by optimizing the collection efficiency of the confocal image by fast z-scanning. However, such images suffer from distortions, particularly in the z dimension, which causes disparities in the x, y, and z directions of the voxels with the original image stacks. As a result, reliable segmentation and feature extraction of these images may be difficult or even impossible. Image interpolation is especially needed for the correction of undersampling artifact in the axial plane of three-dimensional images generated by a confocal microscope to obtain cubic voxels. In this work, we present an adaptive cubic B-spline-based interpolation with the aid of lookup tables by deriving adaptive weights based on local gradients for the sampling nodes in the interpolation formulae. Thus, the proposed method enhances the axial resolution of confocal images by improving the accuracy of the interpolated value simultaneously with great reduction in computational cost. Numerical experimental results confirm the effectiveness of the proposed interpolation approach and demonstrate its superiority both in terms of accuracy and speed compared to other interpolation algorithms.

  15. 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.

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

    PubMed

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

    2013-01-01

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

  17. 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

  18. 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.

  19. Interactive visualization of multiresolution image stacks in 3D.

    PubMed

    Trotts, Issac; Mikula, Shawn; Jones, Edward G

    2007-04-15

    Conventional microscopy, electron microscopy, and imaging techniques such as MRI and PET commonly generate large stacks of images of the sectioned brain. In other domains, such as neurophysiology, variables such as space or time are also varied along a stack axis. Digital image sizes have been progressively increasing and in virtual microscopy, it is now common to work with individual image sizes that are several hundred megapixels and several gigabytes in size. The interactive visualization of these high-resolution, multiresolution images in 2D has been addressed previously [Sullivan, G., and Baker, R., 1994. Efficient quad-tree coding of images and video. IEEE Trans. Image Process. 3 (3), 327-331]. Here, we describe a method for interactive visualization of multiresolution image stacks in 3D. The method, characterized as quad-tree based multiresolution image stack interactive visualization using a texel projection based criterion, relies on accessing and projecting image tiles from multiresolution image stacks in such a way that, from the observer's perspective, image tiles all appear approximately the same size even though they are accessed from different tiers within the images comprising the stack. This method enables efficient navigation of high-resolution image stacks. We implement this method in a program called StackVis, which is a Windows-based, interactive 3D multiresolution image stack visualization system written in C++ and using OpenGL. It is freely available at http://brainmaps.org.

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

    PubMed Central

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

    2009-01-01

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

  1. 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

  2. 3-D imaging of the CNS.

    PubMed

    Runge, V M; Gelblum, D Y; Wood, M L

    1990-01-01

    3-D gradient echo techniques, and in particular FLASH, represent a significant advance in MR imaging strategy allowing thin section, high resolution imaging through a large region of interest. Anatomical areas of application include the brain, spine, and extremities, although the majority of work to date has been performed in the brain. Superior T1 contrast and thus sensitivity to the presence of GdDTPA is achieved with 3-D FLASH when compared to 2-D spin echo technique. There is marked arterial and venous enhancement following Gd DTPA administration on 3-D FLASH, a less common finding with 2-D spin echo. Enhancement of the falx and tentorium is also more prominent. From a single data acquisition, requiring less than 11 min of scan time, high resolution reformatted sagittal, coronal, and axial images can obtained in addition to sections in any arbitrary plane. Tissue segmentation techniques can be applied and lesions displayed in three dimensions. These results may lead to the replacement of 2-D spin echo with 3-D FLASH for high resolution T1-weighted MR imaging of the CNS, particularly in the study of mass lesions and structural anomalies. The application of similar T2-weighted gradient echo techniques may follow, however the signal-to-noise ratio which can be achieved remains a potential limitation.

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

    PubMed

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

    2014-12-15

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

  4. Walker Ranch 3D seismic images

    SciTech Connect

    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.

  5. 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

  6. Backhoe 3D "gold standard" image

    NASA Astrophysics Data System (ADS)

    Gorham, LeRoy; Naidu, Kiranmai D.; Majumder, Uttam; Minardi, Michael A.

    2005-05-01

    ViSUAl-D (VIsual Sar Using ALl Dimensions), a 2004 DARPA/IXO seedling effort, is developing a capability for reliable high confidence ID from standoff ranges. Recent conflicts have demonstrated that the warfighter would greatly benefit from the ability to ID targets beyond visual and electro-optical ranges[1]. Forming optical-quality SAR images while exploiting full polarization, wide angles, and large bandwidth would be key evidence such a capability is achievable. Using data generated by the Xpatch EM scattering code, ViSUAl-D investigates all degrees of freedom available to the radar designer, including 6 GHz bandwidth, full polarization and angle sampling over 2π steradians (upper hemisphere), in order to produce a "literal" image or representation of the target. This effort includes the generation of a "Gold Standard" image that can be produced at X-band utilizing all available target data. This "Gold Standard" image of the backhoe will serve as a test bed for future more relevant military targets and their image development. The seedling team produced a public release data which was released at the 2004 SPIE conference, as well as a 3D "Gold Standard" backhoe image using a 3D image formation algorithm. This paper describes the full backhoe data set, the image formation algorithm, the visualization process and the resulting image.

  7. Tilted planes in 3D image analysis

    NASA Astrophysics Data System (ADS)

    Pargas, Roy P.; Staples, Nancy J.; Malloy, Brian F.; Cantrell, Ken; Chhatriwala, Murtuza

    1998-03-01

    Reliable 3D wholebody scanners which output digitized 3D images of a complete human body are now commercially available. This paper describes a software package, called 3DM, being developed by researchers at Clemson University and which manipulates and extracts measurements from such images. The focus of this paper is on tilted planes, a 3DM tool which allows a user to define a plane through a scanned image, tilt it in any direction, and effectively define three disjoint regions on the image: the points on the plane and the points on either side of the plane. With tilted planes, the user can accurately take measurements required in applications such as apparel manufacturing. The user can manually segment the body rather precisely. Tilted planes assist the user in analyzing the form of the body and classifying the body in terms of body shape. Finally, titled planes allow the user to eliminate extraneous and unwanted points often generated by a 3D scanner. This paper describes the user interface for tilted planes, the equations defining the plane as the user moves it through the scanned image, an overview of the algorithms, and the interaction of the tilted plane feature with other tools in 3DM.

  8. Feasibility of 3D harmonic contrast imaging.

    PubMed

    Voormolen, M M; Bouakaz, A; Krenning, B J; Lancée, C T; ten Cate, F J; de Jong, N

    2004-04-01

    Improved endocardial border delineation with the application of contrast agents should allow for less complex and faster tracing algorithms for left ventricular volume analysis. We developed a fast rotating phased array transducer for 3D imaging of the heart with harmonic capabilities making it suitable for contrast imaging. In this study the feasibility of 3D harmonic contrast imaging is evaluated in vitro. A commercially available tissue mimicking flow phantom was used in combination with Sonovue. Backscatter power spectra from a tissue and contrast region of interest were calculated from recorded radio frequency data. The spectra and the extracted contrast to tissue ratio from these spectra were used to optimize the excitation frequency, the pulse length and the receive filter settings of the transducer. Frequencies ranging from 1.66 to 2.35 MHz and pulse lengths of 1.5, 2 and 2.5 cycles were explored. An increase of more than 15 dB in the contrast to tissue ratio was found around the second harmonic compared with the fundamental level at an optimal excitation frequency of 1.74 MHz and a pulse length of 2.5 cycles. Using the optimal settings for 3D harmonic contrast recordings volume measurements of a left ventricular shaped agar phantom were performed. Without contrast the extracted volume data resulted in a volume error of 1.5%, with contrast an accuracy of 3.8% was achieved. The results show the feasibility of accurate volume measurements from 3D harmonic contrast images. Further investigations will include the clinical evaluation of the presented technique for improved assessment of the heart.

  9. 3D imaging system for biometric applications

    NASA Astrophysics Data System (ADS)

    Harding, Kevin; Abramovich, Gil; Paruchura, Vijay; Manickam, Swaminathan; Vemury, Arun

    2010-04-01

    There is a growing interest in the use of 3D data for many new applications beyond traditional metrology areas. In particular, using 3D data to obtain shape information of both people and objects for applications ranging from identification to game inputs does not require high degrees of calibration or resolutions in the tens of micron range, but does require a means to quickly and robustly collect data in the millimeter range. Systems using methods such as structured light or stereo have seen wide use in measurements, but due to the use of a triangulation angle, and thus the need for a separated second viewpoint, may not be practical for looking at a subject 10 meters away. Even when working close to a subject, such as capturing hands or fingers, the triangulation angle causes occlusions, shadows, and a physically large system that may get in the way. This paper will describe methods to collect medium resolution 3D data, plus highresolution 2D images, using a line of sight approach. The methods use no moving parts and as such are robust to movement (for portability), reliable, and potentially very fast at capturing 3D data. This paper will describe the optical methods considered, variations on these methods, and present experimental data obtained with the approach.

  10. Pattern based 3D image Steganography

    NASA Astrophysics Data System (ADS)

    Thiyagarajan, P.; Natarajan, V.; Aghila, G.; Prasanna Venkatesan, V.; Anitha, R.

    2013-03-01

    This paper proposes a new high capacity Steganographic scheme using 3D geometric models. The novel algorithm re-triangulates a part of a triangle mesh and embeds the secret information into newly added position of triangle meshes. Up to nine bits of secret data can be embedded into vertices of a triangle without causing any changes in the visual quality and the geometric properties of the cover image. Experimental results show that the proposed algorithm is secure, with high capacity and low distortion rate. Our algorithm also resists against uniform affine transformations such as cropping, rotation and scaling. Also, the performance of the method is compared with other existing 3D Steganography algorithms. [Figure not available: see fulltext.

  11. 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.

  12. 3D seismic image processing for interpretation

    NASA Astrophysics Data System (ADS)

    Wu, Xinming

    Extracting fault, unconformity, and horizon surfaces from a seismic image is useful for interpretation of geologic structures and stratigraphic features. Although interpretation of these surfaces has been automated to some extent by others, significant manual effort is still required for extracting each type of these geologic surfaces. I propose methods to automatically extract all the fault, unconformity, and horizon surfaces from a 3D seismic image. To a large degree, these methods just involve image processing or array processing which is achieved by efficiently solving partial differential equations. For fault interpretation, I propose a linked data structure, which is simpler than triangle or quad meshes, to represent a fault surface. In this simple data structure, each sample of a fault corresponds to exactly one image sample. Using this linked data structure, I extract complete and intersecting fault surfaces without holes from 3D seismic images. I use the same structure in subsequent processing to estimate fault slip vectors. I further propose two methods, using precomputed fault surfaces and slips, to undo faulting in seismic images by simultaneously moving fault blocks and faults themselves. For unconformity interpretation, I first propose a new method to compute a unconformity likelihood image that highlights both the termination areas and the corresponding parallel unconformities and correlative conformities. I then extract unconformity surfaces from the likelihood image and use these surfaces as constraints to more accurately estimate seismic normal vectors that are discontinuous near the unconformities. Finally, I use the estimated normal vectors and use the unconformities as constraints to compute a flattened image, in which seismic reflectors are all flat and vertical gaps correspond to the unconformities. Horizon extraction is straightforward after computing a map of image flattening; we can first extract horizontal slices in the flattened space

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

    NASA Astrophysics Data System (ADS)

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

    2006-02-01

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

  14. 3D GPR Imaging of Wooden Logs

    NASA Astrophysics Data System (ADS)

    Halabe, Udaya B.; Pyakurel, Sandeep

    2007-03-01

    There has been a lack of an effective NDE technique to locate internal defects within wooden logs. The few available elastic wave propagation based techniques are limited to predicting E values. Other techniques such as X-rays have not been very successful in detecting internal defects in logs. If defects such as embedded metals could be identified before the sawing process, the saw mills could significantly increase their production by reducing the probability of damage to the saw blade and the associated downtime and the repair cost. Also, if the internal defects such as knots and decayed areas could be identified in logs, the sawing blade can be oriented to exclude the defective portion and optimize the volume of high valued lumber that can be obtained from the logs. In this research, GPR has been successfully used to locate internal defects (knots, decays and embedded metals) within the logs. This paper discusses GPR imaging and mapping of the internal defects using both 2D and 3D interpretation methodology. Metal pieces were inserted in a log and the reflection patterns from these metals were interpreted from the radargrams acquired using 900 MHz antenna. Also, GPR was able to accurately identify the location of knots and decays. Scans from several orientations of the log were collected to generate 3D cylindrical volume. The actual location of the defects showed good correlation with the interpreted defects in the 3D volume. The time/depth slices from 3D cylindrical volume data were useful in understanding the extent of defects inside the log.

  15. 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.

  16. 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.

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

    PubMed

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

    2014-12-01

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

  18. 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.

  19. 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.

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

    PubMed

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

    2016-06-01

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

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

    PubMed

    Koga, Daisuke; Ushiki, Tatsuo; Watanabe, Tsuyoshi

    2017-01-01

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

  2. [3D display of sequential 2D medical images].

    PubMed

    Lu, Yisong; Chen, Yazhu

    2003-12-01

    A detailed review is given in this paper on various current 3D display methods for sequential 2D medical images and the new development in 3D medical image display. True 3D display, surface rendering, volume rendering, 3D texture mapping and distributed collaborative rendering are discussed in depth. For two kinds of medical applications: Real-time navigation system and high-fidelity diagnosis in computer aided surgery, different 3D display methods are presented.

  3. 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.

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

    PubMed Central

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

    2015-01-01

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

  5. Infrastructure for 3D Imaging Test Bed

    DTIC Science & Technology

    2007-05-11

    analysis. (c.) Real time detection & analysis of human gait: using a video camera we capture walking human silhouette for pattern modeling and gait ... analysis . Fig. 5 shows the scanning result result that is fed into a Geo-magic software tool for 3D meshing. Fig. 5: 3D scanning result In

  6. 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.

  7. 3D Gabor wavelet based vessel filtering of photoacoustic images.

    PubMed

    Haq, Israr Ul; Nagoaka, Ryo; Makino, Takahiro; Tabata, Takuya; Saijo, Yoshifumi

    2016-08-01

    Filtering and segmentation of vasculature is an important issue in medical imaging. The visualization of vasculature is crucial for the early diagnosis and therapy in numerous medical applications. This paper investigates the use of Gabor wavelet to enhance the effect of vasculature while eliminating the noise due to size, sensitivity and aperture of the detector in 3D Optical Resolution Photoacoustic Microscopy (OR-PAM). A detailed multi-scale analysis of wavelet filtering and Hessian based method is analyzed for extracting vessels of different sizes since the blood vessels usually vary with in a range of radii. The proposed algorithm first enhances the vasculature in the image and then tubular structures are classified by eigenvalue decomposition of the local Hessian matrix at each voxel in the image. The algorithm is tested on non-invasive experiments, which shows appreciable results to enhance vasculature in photo-acoustic images.

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

    PubMed

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

    2015-06-01

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

  9. Glasses-free 3D viewing systems for medical imaging

    NASA Astrophysics Data System (ADS)

    Magalhães, Daniel S. F.; Serra, Rolando L.; Vannucci, André L.; Moreno, Alfredo B.; Li, Li M.

    2012-04-01

    In this work we show two different glasses-free 3D viewing systems for medical imaging: a stereoscopic system that employs a vertically dispersive holographic screen (VDHS) and a multi-autostereoscopic system, both used to produce 3D MRI/CT images. We describe how to obtain a VDHS in holographic plates optimized for this application, with field of view of 7 cm to each eye and focal length of 25 cm, showing images done with the system. We also describe a multi-autostereoscopic system, presenting how it can generate 3D medical imaging from viewpoints of a MRI or CT image, showing results of a 3D angioresonance image.

  10. 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

  11. Experiments on terahertz 3D scanning microscopic imaging

    NASA Astrophysics Data System (ADS)

    Zhou, Yi; Li, Qi

    2016-10-01

    Compared with the visible light and infrared, terahertz (THz) radiation can penetrate nonpolar and nonmetallic materials. There are many studies on the THz coaxial transmission confocal microscopy currently. But few researches on the THz dual-axis reflective confocal microscopy were reported. In this paper, we utilized a dual-axis reflective confocal scanning microscope working at 2.52 THz. In contrast with the THz coaxial transmission confocal microscope, the microscope adopted in this paper can attain higher axial resolution at the expense of reduced lateral resolution, revealing more satisfying 3D imaging capability. Objects such as Chinese characters "Zhong-Hua" written in paper with a pencil and a combined sheet metal which has three layers were scanned. The experimental results indicate that the system can extract two Chinese characters "Zhong," "Hua" or three layers of the combined sheet metal. It can be predicted that the microscope can be applied to biology, medicine and other fields in the future due to its favorable 3D imaging capability.

  12. 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

  13. 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.

  14. Research of range-gated 3D imaging technology

    NASA Astrophysics Data System (ADS)

    Yang, Haitao; Zhao, Hongli; Youchen, Fan

    2016-10-01

    Laser image data-based target recognition technology is one of the key technologies of laser active imaging systems. This paper discussed the status quo of 3-D imaging development at home and abroad, analyzed the current technological bottlenecks, and built a prototype of range-gated systems to obtain a set of range-gated slice images, and then constructed the 3-D images of the target by binary method and centroid method, respectively, and by constructing different numbers of slice images explored the relationship between the number of images and the reconstruction accuracy in the 3-D image reconstruction process. The experiment analyzed the impact of two algorithms, binary method and centroid method, on the results of 3-D image reconstruction. In the binary method, a comparative analysis was made on the impact of different threshold values on the results of reconstruction, where 0.1, 0.2, 0.3 and adaptive threshold values were selected for 3-D reconstruction of the slice images. In the centroid method, 15, 10, 6, 3, and 2 images were respectively used to realize 3-D reconstruction. Experimental results showed that with the same number of slice images, the accuracy of centroid method was higher than the binary algorithm, and the binary algorithm had a large dependence on the selection of threshold; with the number of slice images dwindling, the accuracy of images reconstructed by centroid method continued to reduce, and at least three slice images were required in order to obtain one 3-D image.

  15. 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

  16. Image Force Microscopy

    NASA Astrophysics Data System (ADS)

    Rajapaksa, Indrajith

    In this thesis we describe an enhancement to the Atomic force microscope (AFM) to simultaneously gather topographic features and spectroscopic information .Compared to the current state of the art of near-field excitation and far-field detection AFM imaging techniques our system uses a radical new approach near-field excitation and near-field detection. By placing the detector in the near-field we achieve high signal to noise and single molecular resolution. The origin of our near-field detector signal is the image force gradient due to the interaction of the stimulated molecular dipole with its image on the metal probe. We designed and built an optical and electronic system to capture this signal and simultaneously image nano-scale surface topography and optical image force gradient. By varying the wavelength of the excitation beam we measure the induced optical image force gradient spectra of molecules on surface. These spectra show good agreement with the absorption spectra of the bulk molecules measured by conventional absorption spectroscopy. We show that image force gradient is directly proportional to the optical absorption dipole strength. Using Finite Element 3D electromagnetic simulations and using Lorentz model for the excited molecular dipole we showed that the image force gradient has a decay length of 1nm, making the theoretical resolution of this microscopy technique approximately 1 nm. This rapid decay was measured experimentally .This resolution was seen by the high contrasting spectroscopic images of molecules on the surface. In follow on experiments this technique was extended to provide surface Raman spectroscopy and microscopy at molecular resolution. We create an image force gradient interaction through optical parametric down conversion between stimulated Raman excited molecules on a surface and a cantilevered nanometer scale probe brought very close to it. Spectroscopy and microscopy on clusters of molecules have been performed. Single

  17. Reconstruction-based 3D/2D image registration.

    PubMed

    Tomazevic, Dejan; Likar, Bostjan; Pernus, Franjo

    2005-01-01

    In this paper we present a novel 3D/2D registration method, where first, a 3D image is reconstructed from a few 2D X-ray images and next, the preoperative 3D image is brought into the best possible spatial correspondence with the reconstructed image by optimizing a similarity measure. Because the quality of the reconstructed image is generally low, we introduce a novel asymmetric mutual information similarity measure, which is able to cope with low image quality as well as with different imaging modalities. The novel 3D/2D registration method has been evaluated using standardized evaluation methodology and publicly available 3D CT, 3DRX, and MR and 2D X-ray images of two spine phantoms, for which gold standard registrations were known. In terms of robustness, reliability and capture range the proposed method outperformed the gradient-based method and the method based on digitally reconstructed radiographs (DRRs).

  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 Imaging with Holographic Tomography

    NASA Astrophysics Data System (ADS)

    Sheppard, Colin J. R.; Kou, Shan Shan

    2010-04-01

    There are two main types of tomography that enable the 3D internal structures of objects to be reconstructed from scattered data. The commonly known computerized tomography (CT) give good results in the x-ray wavelength range where the filtered back-projection theorem and Radon transform can be used. These techniques rely on the Fourier projection-slice theorem where rays are considered to propagate straight through the object. Another type of tomography called `diffraction tomography' applies in applications in optics and acoustics where diffraction and scattering effects must be taken into account. The latter proves to be a more difficult problem, as light no longer travels straight through the sample. Holographic tomography is a popular way of performing diffraction tomography and there has been active experimental research on reconstructing complex refractive index data using this approach recently. However, there are two distinct ways of doing tomography: either by rotation of the object or by rotation of the illumination while fixing the detector. The difference between these two setups is intuitive but needs to be quantified. From Fourier optics and information transformation point of view, we use 3D transfer function analysis to quantitatively describe how spatial frequencies of the object are mapped to the Fourier domain. We first employ a paraxial treatment by calculating the Fourier transform of the defocused OTF. The shape of the calculated 3D CTF for tomography, by scanning the illumination in one direction only, takes on a form that we might call a 'peanut,' compared to the case of object rotation, where a diablo is formed, the peanut exhibiting significant differences and non-isotropy. In particular, there is a line singularity along one transverse direction. Under high numerical aperture conditions, the paraxial treatment is not accurate, and so we make use of 3D analytical geometry to calculate the behaviour in the non-paraxial case. This time, we

  20. 3-D imaging and illustration of mouse intestinal neurovascular complex.

    PubMed

    Fu, Ya-Yuan; Peng, Shih-Jung; Lin, Hsin-Yao; Pasricha, Pankaj J; Tang, Shiue-Cheng

    2013-01-01

    Because of the dispersed nature of nerves and blood vessels, standard histology cannot provide a global and associated observation of the enteric nervous system (ENS) and vascular network. We prepared transparent mouse intestine and combined vessel painting and three-dimensional (3-D) neurohistology for joint visualization of the ENS and vasculature. Cardiac perfusion of the fluorescent wheat germ agglutinin (vessel painting) was used to label the ileal blood vessels. The pan-neuronal marker PGP9.5, sympathetic neuronal marker tyrosine hydroxylase (TH), serotonin, and glial markers S100B and GFAP were used as the immunostaining targets of neural tissues. The fluorescently labeled specimens were immersed in the optical clearing solution to improve photon penetration for 3-D confocal microscopy. Notably, we simultaneously revealed the ileal microstructure, vasculature, and innervation with micrometer-level resolution. Four examples are given: 1) the morphology of the TH-labeled sympathetic nerves: sparse in epithelium, perivascular at the submucosa, and intraganglionic at myenteric plexus; 2) distinct patterns of the extrinsic perivascular and intrinsic pericryptic innervation at the submucosal-mucosal interface; 3) different associations of serotonin cells with the mucosal neurovascular elements in the villi and crypts; and 4) the periganglionic capillary network at the myenteric plexus and its contact with glial fibers. Our 3-D imaging approach provides a useful tool to simultaneously reveal the nerves and blood vessels in a space continuum for panoramic illustration and analysis of the neurovascular complex to better understand the intestinal physiology and diseases.

  1. Optical 3D imaging and visualization of concealed objects

    NASA Astrophysics Data System (ADS)

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

    2016-09-01

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

  2. 3D reconstruction of cortical microtubules using multi-angle total internal reflection fluorescence microscopy

    NASA Astrophysics Data System (ADS)

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

    2017-01-01

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

  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 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

  5. 3D X-ray ultra-microscopy of bone tissue.

    PubMed

    Langer, M; Peyrin, F

    2016-02-01

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

  6. 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.

  7. 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.

  8. Volumetric image display for complex 3D data visualization

    NASA Astrophysics Data System (ADS)

    Tsao, Che-Chih; Chen, Jyh Shing

    2000-05-01

    A volumetric image display is a new display technology capable of displaying computer generated 3D images in a volumetric space. Many viewers can walk around the display and see the image from omni-directions simultaneously without wearing any glasses. The image is real and possesses all major elements in both physiological and psychological depth cues. Due to the volumetric nature of its image, the VID can provide the most natural human-machine interface in operations involving 3D data manipulation and 3D targets monitoring. The technology creates volumetric 3D images by projecting a series of profiling images distributed in the space form a volumetric image because of the after-image effect of human eyes. Exemplary applications in biomedical image visualization were tested on a prototype display, using different methods to display a data set from Ct-scans. The features of this display technology make it most suitable for applications that require quick understanding of the 3D relations, need frequent spatial interactions with the 3D images, or involve time-varying 3D data. It can also be useful for group discussion and decision making.

  9. 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.

  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. 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.

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

    PubMed Central

    Rane, Tushar D.; Armani, Andrea M.

    2016-01-01

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

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

    PubMed

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

    2014-08-01

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

  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. 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.

  16. Quantitative Multiscale Cell Imaging in Controlled 3D Microenvironments

    PubMed Central

    Welf, Erik S.; Driscoll, Meghan K.; Dean, Kevin M.; Schäfer, Claudia; Chu, Jun; Davidson, Michael W.; Lin, Michael Z.; Danuser, Gaudenz; Fiolka, Reto

    2016-01-01

    The microenvironment determines cell behavior, but the underlying molecular mechanisms are poorly understood because quantitative studies of cell signaling and behavior have been challenging due to insufficient spatial and/or temporal resolution and limitations on microenvironmental control. Here we introduce microenvironmental selective plane illumination microscopy (meSPIM) for imaging and quantification of intracellular signaling and submicrometer cellular structures as well as large-scale cell morphological and environmental features. We demonstrate the utility of this approach by showing that the mechanical properties of the microenvironment regulate the transition of melanoma cells from actin-driven protrusion to blebbing, and we present tools to quantify how cells manipulate individual collagen fibers. We leverage the nearly isotropic resolution of meSPIM to quantify the local concentration of actin and phosphatidylinositol 3-kinase signaling on the surfaces of cells deep within 3D collagen matrices and track the many small membrane protrusions that appear in these more physiologically relevant environments. PMID:26906741

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

    PubMed

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

    2016-11-01

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

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. Seeing More Is Knowing More: V3D Enables Real-Time 3D Visualization and Quantitative Analysis of Large-Scale Biological Image Data Sets

    NASA Astrophysics Data System (ADS)

    Peng, Hanchuan; Long, Fuhui

    Everyone understands seeing more is knowing more. However, for large-scale 3D microscopic image analysis, it has not been an easy task to efficiently visualize, manipulate and understand high-dimensional data in 3D, 4D or 5D spaces. We developed a new 3D+ image visualization and analysis platform, V3D, to meet this need. The V3D system provides 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 V3Dbased 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 high-resolution 3D digital atlas of neurite tracts in the fruitfly brain. V3D can be easily extended using a simple-to-use and comprehensive plugin interface.

  3. 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.

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

    PubMed

    Carnicer, Artur; Javidi, Bahram

    2015-03-09

    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.

  5. 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.

  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. 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.

  8. 3-D Imaging Based, Radiobiological Dosimetry

    PubMed Central

    Sgouros, George; Frey, Eric; Wahl, Richard; He, Bin; Prideaux, Andrew; Hobbs, Robert

    2008-01-01

    Targeted radionuclide therapy holds promise as a new treatment against cancer. Advances in imaging are making it possible to evaluate the spatial distribution of radioactivity in tumors and normal organs over time. Matched anatomical imaging such as combined SPECT/CT and PET/CT have also made it possible to obtain tissue density information in conjunction with the radioactivity distribution. Coupled with sophisticated iterative reconstruction algorithims, these advances have made it possible to perform highly patient-specific dosimetry that also incorporates radiobiological modeling. Such sophisticated dosimetry techniques are still in the research investigation phase. Given the attendant logistical and financial costs, a demonstrated improvement in patient care will be a prerequisite for the adoption of such highly-patient specific internal dosimetry methods. PMID:18662554

  9. A 3D Level Set Method for Microwave Breast Imaging

    PubMed Central

    Colgan, Timothy J.; Hagness, Susan C.; Van Veen, Barry D.

    2015-01-01

    Objective Conventional inverse-scattering algorithms for microwave breast imaging result in moderate resolution images with blurred boundaries between tissues. Recent 2D numerical microwave imaging studies demonstrate that the use of a level set method preserves dielectric boundaries, resulting in a more accurate, higher resolution reconstruction of the dielectric properties distribution. Previously proposed level set algorithms are computationally expensive and thus impractical in 3D. In this paper we present a computationally tractable 3D microwave imaging algorithm based on level sets. Methods We reduce the computational cost of the level set method using a Jacobian matrix, rather than an adjoint method, to calculate Frechet derivatives. We demonstrate the feasibility of 3D imaging using simulated array measurements from 3D numerical breast phantoms. We evaluate performance by comparing full 3D reconstructions to those from a conventional microwave imaging technique. We also quantitatively assess the efficacy of our algorithm in evaluating breast density. Results Our reconstructions of 3D numerical breast phantoms improve upon those of a conventional microwave imaging technique. The density estimates from our level set algorithm are more accurate than those of conventional microwave imaging, and the accuracy is greater than that reported for mammographic density estimation. Conclusion Our level set method leads to a feasible level of computational complexity for full 3D imaging, and reconstructs the heterogeneous dielectric properties distribution of the breast more accurately than conventional microwave imaging methods. Significance 3D microwave breast imaging using a level set method is a promising low-cost, non-ionizing alternative to current breast imaging techniques. PMID:26011863

  10. 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.

  11. 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.

  12. Potential Cost Savings with 3D Printing Combined With 3D Imaging and CPLM for Fleet Maintenance and Revitalization

    DTIC Science & Technology

    2014-05-01

    1 Potential Cost Savings with 3D Printing Combined With 3D Imaging and CPLM for Fleet Maintenance and Revitalization David N. Ford...2014 4. TITLE AND SUBTITLE Potential Cost Savings with 3D Printing Combined With 3D Imaging and CPLM for Fleet Maintenance and Revitalization 5a...Manufacturing ( 3D printing ) 2 Research Context Problem: Learning curve savings forecasted in SHIPMAIN maintenance initiative have not materialized

  13. Morphometrics, 3D Imaging, and Craniofacial Development

    PubMed Central

    Hallgrimsson, Benedikt; Percival, Christopher J.; Green, Rebecca; Young, Nathan M.; Mio, Washington; Marcucio, Ralph

    2017-01-01

    Recent studies have shown how volumetric imaging and morphometrics can add significantly to our understanding of morphogenesis, the developmental basis for variation and the etiology of structural birth defects. On the other hand, the complex questions and diverse imaging data in developmental biology present morphometrics with more complex challenges than applications in virtually any other field. Meeting these challenges is necessary in order to understand the mechanistic basis for variation in complex morphologies. This chapter reviews the methods and theory that enable the application of modern landmark-based morphometrics to developmental biology and craniofacial development, in particular. We discuss the theoretical foundations of morphometrics as applied to development and review the basic approaches to the quantification of morphology. Focusing on geometric morphometrics, we discuss the principal statistical methods for quantifying and comparing morphological variation and covariation structure within and among groups. Finally, we discuss the future directions for morphometrics in developmental biology that will be required for approaches that enable quantitative integration across the genotype-phenotype map. PMID:26589938

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

    PubMed

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

    2017-03-20

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

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

    NASA Astrophysics Data System (ADS)

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

    2017-01-01

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

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

    PubMed Central

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

    2017-01-01

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

  17. 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.

  18. 3D Whole Heart Imaging for Congenital Heart Disease

    PubMed Central

    Greil, Gerald; Tandon, Animesh (Aashoo); Silva Vieira, Miguel; Hussain, Tarique

    2017-01-01

    Three-dimensional (3D) whole heart techniques form a cornerstone in cardiovascular magnetic resonance imaging of congenital heart disease (CHD). It offers significant advantages over other CHD imaging modalities and techniques: no ionizing radiation; ability to be run free-breathing; ECG-gated dual-phase imaging for accurate measurements and tissue properties estimation; and higher signal-to-noise ratio and isotropic voxel resolution for multiplanar reformatting assessment. However, there are limitations, such as potentially long acquisition times with image quality degradation. Recent advances in and current applications of 3D whole heart imaging in CHD are detailed, as well as future directions. PMID:28289674

  19. 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

  20. A colour image reproduction framework for 3D colour printing

    NASA Astrophysics Data System (ADS)

    Xiao, Kaida; Sohiab, Ali; Sun, Pei-li; Yates, Julian M.; Li, Changjun; Wuerger, Sophie

    2016-10-01

    In this paper, the current technologies in full colour 3D printing technology were introduced. A framework of colour image reproduction process for 3D colour printing is proposed. A special focus was put on colour management for 3D printed objects. Two approaches, colorimetric colour reproduction and spectral based colour reproduction are proposed in order to faithfully reproduce colours in 3D objects. Two key studies, colour reproduction for soft tissue prostheses and colour uniformity correction across different orientations are described subsequently. Results are clear shown that applying proposed colour image reproduction framework, performance of colour reproduction can be significantly enhanced. With post colour corrections, a further improvement in colour process are achieved for 3D printed objects.

  1. 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.

  2. 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

  3. Digital holography and 3D imaging: introduction to feature issue.

    PubMed

    Kim, Myung K; Hayasaki, Yoshio; Picart, Pascal; Rosen, Joseph

    2013-01-01

    This feature issue of Applied Optics on Digital Holography and 3D Imaging is the sixth of an approximately annual series. Forty-seven papers are presented, covering a wide range of topics in phase-shifting methods, low coherence methods, particle analysis, biomedical imaging, computer-generated holograms, integral imaging, and many others.

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

    PubMed

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

    2004-05-18

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

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

    NASA Astrophysics Data System (ADS)

    Li, Xiao Wei; Kim, Seok Tae

    2013-12-01

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

  6. Progresses in 3D integral imaging with optical processing

    NASA Astrophysics Data System (ADS)

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

    2008-11-01

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

  7. DCT and DST Based Image Compression for 3D Reconstruction

    NASA Astrophysics Data System (ADS)

    Siddeq, Mohammed M.; Rodrigues, Marcos A.

    2017-03-01

    This paper introduces a new method for 2D image compression whose quality is demonstrated through accurate 3D reconstruction using structured light techniques and 3D reconstruction from multiple viewpoints. The method is based on two discrete transforms: (1) A one-dimensional Discrete Cosine Transform (DCT) is applied to each row of the image. (2) The output from the previous step is transformed again by a one-dimensional Discrete Sine Transform (DST), which is applied to each column of data generating new sets of high-frequency components followed by quantization of the higher frequencies. The output is then divided into two parts where the low-frequency components are compressed by arithmetic coding and the high frequency ones by an efficient minimization encoding algorithm. At decompression stage, a binary search algorithm is used to recover the original high frequency components. The technique is demonstrated by compressing 2D images up to 99% compression ratio. The decompressed images, which include images with structured light patterns for 3D reconstruction and from multiple viewpoints, are of high perceptual quality yielding accurate 3D reconstruction. Perceptual assessment and objective quality of compression are compared with JPEG and JPEG2000 through 2D and 3D RMSE. Results show that the proposed compression method is superior to both JPEG and JPEG2000 concerning 3D reconstruction, and with equivalent perceptual quality to JPEG2000.

  8. 3D Subharmonic Ultrasound Imaging In Vitro and In Vivo

    PubMed Central

    Eisenbrey, John R.; Sridharan, Anush; Machado, Priscilla; Zhao, Hongjia; Halldorsdottir, Valgerdur G.; Dave, Jaydev K.; Liu, Ji-Bin; Park, Suhyun; Dianis, Scott; Wallace, Kirk; Thomenius, Kai E.; Forsberg, F.

    2012-01-01

    Rationale and Objectives While contrast-enhanced ultrasound imaging techniques such as harmonic imaging (HI) have evolved to reduce tissue signals using the nonlinear properties of the contrast agent, levels of background suppression have been mixed. Subharmonic imaging (SHI) offers near-complete tissue suppression by centering the receive bandwidth at half the transmitting frequency. In this work we demonstrate the feasibility of 3D SHI and compare it to 3D HI. Materials and Methods 3D HI and SHI were implemented on a Logiq 9 ultrasound scanner (GE Healthcare, Milwaukee, Wisconsin) with a 4D10L probe. Four-cycle SHI was implemented to transmit at 5.8 MHz and receive at 2.9 MHz, while 2-cycle HI was implemented to transmit at 5 MHz and receive at 10 MHz. The ultrasound contrast agent Definity (Lantheus Medical Imaging, North Billerica, MA) was imaged within a flow phantom and the lower pole of two canine kidneys in both HI and SHI modes. Contrast to tissue ratios (CTR) and rendered images were compared offline. Results SHI resulted in significant improvement in CTR levels relative to HI both in vitro (12.11±0.52 vs. 2.67±0.77, p<0.001) and in vivo (5.74±1.92 vs. 2.40±0.48, p=0.04). Rendered 3D SHI images provided better tissue suppression and a greater overall view of vessels in a flow phantom and canine renal vasculature. Conclusions The successful implementation of SHI in 3D allows imaging of vascular networks over a heterogeneous sample volume and should improve future diagnostic accuracy. Additionally, 3D SHI provides improved CTR values relative to 3D HI. PMID:22464198

  9. Low Dose, Low Energy 3d Image Guidance during Radiotherapy

    NASA Astrophysics Data System (ADS)

    Moore, C. J.; Marchant, T.; Amer, A.; Sharrock, P.; Price, P.; Burton, D.

    2006-04-01

    Patient kilo-voltage X-ray cone beam volumetric imaging for radiotherapy was first demonstrated on an Elekta Synergy mega-voltage X-ray linear accelerator. Subsequently low dose, reduced profile reconstruction imaging was shown to be practical for 3D geometric setup registration to pre-treatment planning images without compromising registration accuracy. Reconstruction from X-ray profiles gathered between treatment beam deliveries was also introduced. The innovation of zonal cone beam imaging promises significantly reduced doses to patients and improved soft tissue contrast in the tumour target zone. These developments coincided with the first dynamic 3D monitoring of continuous body topology changes in patients, at the moment of irradiation, using a laser interferometer. They signal the arrival of low dose, low energy 3D image guidance during radiotherapy itself.

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

    PubMed Central

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

    2016-01-01

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

  11. Accelerated 3D catheter visualization from triplanar MR projection images.

    PubMed

    Schirra, Carsten Oliver; Weiss, Steffen; Krueger, Sascha; Caulfield, Denis; Pedersen, Steen F; Razavi, Reza; Kozerke, Sebastian; Schaeffter, Tobias

    2010-07-01

    One major obstacle for MR-guided catheterizations is long acquisition times associated with visualizing interventional devices. Therefore, most techniques presented hitherto rely on single-plane imaging to visualize the catheter. Recently, accelerated three-dimensional (3D) imaging based on compressed sensing has been proposed to reduce acquisition times. However, frame rates with this technique remain low, and the 3D reconstruction problem yields a considerable computational load. In X-ray angiography, it is well understood that the shape of interventional devices can be derived in 3D space from a limited number of projection images. In this work, this fact is exploited to develop a method for 3D visualization of active catheters from multiplanar two-dimensional (2D) projection MR images. This is favorable to 3D MRI as the overall number of acquired profiles, and consequently the acquisition time, is reduced. To further reduce measurement times, compressed sensing is employed. Furthermore, a novel single-channel catheter design is presented that combines a solenoidal tip coil in series with a single-loop antenna, enabling simultaneous tip tracking and shape visualization. The tracked tip and catheter properties provide constraints for compressed sensing reconstruction and subsequent 2D/3D curve fitting. The feasibility of the method is demonstrated in phantoms and in an in vivo pig experiment.

  12. Prostate Mechanical Imaging: 3-D Image Composition and Feature Calculations

    PubMed Central

    Egorov, Vladimir; Ayrapetyan, Suren; Sarvazyan, Armen P.

    2008-01-01

    We have developed a method and a device entitled prostate mechanical imager (PMI) for the real-time imaging of prostate using a transrectal probe equipped with a pressure sensor array and position tracking sensor. PMI operation is based on measurement of the stress pattern on the rectal wall when the probe is pressed against the prostate. Temporal and spatial changes in the stress pattern provide information on the elastic structure of the gland and allow two-dimensional (2-D) and three-dimensional (3-D) reconstruction of prostate anatomy and assessment of prostate mechanical properties. The data acquired allow the calculation of prostate features such as size, shape, nodularity, consistency/hardness, and mobility. The PMI prototype has been validated in laboratory experiments on prostate phantoms and in a clinical study. The results obtained on model systems and in vivo images from patients prove that PMI has potential to become a diagnostic tool that could largely supplant DRE through its higher sensitivity, quantitative record storage, ease-of-use and inherent low cost. PMID:17024836

  13. Exposing digital image forgeries by 3D reconstruction technology

    NASA Astrophysics Data System (ADS)

    Wang, Yongqiang; Xu, Xiaojing; Li, Zhihui; Liu, Haizhen; Li, Zhigang; Huang, Wei

    2009-11-01

    Digital images are easy to tamper and edit due to availability of powerful image processing and editing software. Especially, forged images by taking from a picture of scene, because of no manipulation was made after taking, usual methods, such as digital watermarks, statistical correlation technology, can hardly detect the traces of image tampering. According to image forgery characteristics, a method, based on 3D reconstruction technology, which detect the forgeries by discriminating the dimensional relationship of each object appeared on image, is presented in this paper. This detection method includes three steps. In the first step, all the parameters of images were calibrated and each crucial object on image was chosen and matched. In the second step, the 3D coordinates of each object were calculated by bundle adjustment. In final step, the dimensional relationship of each object was analyzed. Experiments were designed to test this detection method; the 3D reconstruction and the forged image 3D reconstruction were computed independently. Test results show that the fabricating character in digital forgeries can be identified intuitively by this method.

  14. Building 3D scenes from 2D image sequences

    NASA Astrophysics Data System (ADS)

    Cristea, Paul D.

    2006-05-01

    Sequences of 2D images, taken by a single moving video receptor, can be fused to generate a 3D representation. This dynamic stereopsis exists in birds and reptiles, whereas the static binocular stereopsis is common in mammals, including humans. Most multimedia computer vision systems for stereo image capture, transmission, processing, storage and retrieval are based on the concept of binocularity. As a consequence, their main goal is to acquire, conserve and enhance pairs of 2D images able to generate a 3D visual perception in a human observer. Stereo vision in birds is based on the fusion of images captured by each eye, with previously acquired and memorized images from the same eye. The process goes on simultaneously and conjointly for both eyes and generates an almost complete all-around visual field. As a consequence, the baseline distance is no longer fixed, as in the case of binocular 3D view, but adjustable in accordance with the distance to the object of main interest, allowing a controllable depth effect. Moreover, the synthesized 3D scene can have a better resolution than each individual 2D image in the sequence. Compression of 3D scenes can be achieved, and stereo transmissions with lower bandwidth requirements can be developed.

  15. 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.

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

    PubMed

    Belichenko, P V; Dahlström, A

    1995-09-01

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

  17. 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.

  18. Visualization and analysis of 3D microscopic images.

    PubMed

    Long, Fuhui; Zhou, Jianlong; Peng, Hanchuan

    2012-01-01

    In a wide range of biological studies, it is highly desirable to visualize and analyze three-dimensional (3D) microscopic images. In this primer, we first introduce several major methods for visualizing typical 3D images and related multi-scale, multi-time-point, multi-color data sets. Then, we discuss three key categories of image analysis tasks, namely segmentation, registration, and annotation. We demonstrate how to pipeline these visualization and analysis modules using examples of profiling the single-cell gene-expression of C. elegans and constructing a map of stereotyped neurite tracts in a fruit fly brain.

  19. Visualization and Analysis of 3D Microscopic Images

    PubMed Central

    Long, Fuhui; Zhou, Jianlong; Peng, Hanchuan

    2012-01-01

    In a wide range of biological studies, it is highly desirable to visualize and analyze three-dimensional (3D) microscopic images. In this primer, we first introduce several major methods for visualizing typical 3D images and related multi-scale, multi-time-point, multi-color data sets. Then, we discuss three key categories of image analysis tasks, namely segmentation, registration, and annotation. We demonstrate how to pipeline these visualization and analysis modules using examples of profiling the single-cell gene-expression of C. elegans and constructing a map of stereotyped neurite tracts in a fruit fly brain. PMID:22719236

  20. 3D Image Reconstruction: Determination of Pattern Orientation

    SciTech Connect

    Blankenbecler, Richard

    2003-03-13

    The problem of determining the euler angles of a randomly oriented 3-D object from its 2-D Fraunhofer diffraction patterns is discussed. This problem arises in the reconstruction of a positive semi-definite 3-D object using oversampling techniques. In such a problem, the data consists of a measured set of magnitudes from 2-D tomographic images of the object at several unknown orientations. After the orientation angles are determined, the object itself can then be reconstructed by a variety of methods using oversampling, the magnitude data from the 2-D images, physical constraints on the image and then iteration to determine the phases.

  1. Accuracy of 3D Imaging Software in Cephalometric Analysis

    DTIC Science & Technology

    2013-06-21

    Imaging and Communication in Medicine ( DICOM ) files into personal computer-based software to enable 3D reconstruction of the craniofacial skeleton. These...tissue profile. CBCT data can be imported as DICOM files into personal computer–based software to provide 3D reconstruction of the craniofacial...been acquired for the three pig models. The CBCT data were exported into DICOM multi-file format. They will be imported into a proprietary

  2. 3D Image Display Courses for Information Media Students.

    PubMed

    Yanaka, Kazuhisa; Yamanouchi, Toshiaki

    2016-01-01

    Three-dimensional displays are used extensively in movies and games. These displays are also essential in mixed reality, where virtual and real spaces overlap. Therefore, engineers and creators should be trained to master 3D display technologies. For this reason, the Department of Information Media at the Kanagawa Institute of Technology has launched two 3D image display courses specifically designed for students who aim to become information media engineers and creators.

  3. 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.

  4. Gastric Contraction Imaging System Using a 3-D Endoscope.

    PubMed

    Yoshimoto, Kayo; Yamada, Kenji; Watabe, Kenji; Takeda, Maki; Nishimura, Takahiro; Kido, Michiko; Nagakura, Toshiaki; Takahashi, Hideya; Nishida, Tsutomu; Iijima, Hideki; Tsujii, Masahiko; Takehara, Tetsuo; Ohno, Yuko

    2014-01-01

    This paper presents a gastric contraction imaging system for assessment of gastric motility using a 3-D endoscope. Gastrointestinal diseases are mainly based on morphological abnormalities. However, gastrointestinal symptoms are sometimes apparent without visible abnormalities. One of the major factors for these diseases is abnormal gastrointestinal motility. For assessment of gastric motility, a gastric motility imaging system is needed. To assess the dynamic motility of the stomach, the proposed system measures 3-D gastric contractions derived from a 3-D profile of the stomach wall obtained with a developed 3-D endoscope. After obtaining contraction waves, their frequency, amplitude, and speed of propagation can be calculated using a Gaussian function. The proposed system was evaluated for 3-D measurements of several objects with known geometries. The results showed that the surface profiles could be obtained with an error of [Formula: see text] of the distance between two different points on images. Subsequently, we evaluated the validity of a prototype system using a wave simulated model. In the experiment, the amplitude and position of waves could be measured with 1-mm accuracy. The present results suggest that the proposed system can measure the speed and amplitude of contractions. This system has low invasiveness and can assess the motility of the stomach wall directly in a 3-D manner. Our method can be used for examination of gastric morphological and functional abnormalities.

  5. 2D/3D Image Registration using Regression Learning

    PubMed Central

    Chou, Chen-Rui; Frederick, Brandon; Mageras, Gig; Chang, Sha; Pizer, Stephen

    2013-01-01

    In computer vision and image analysis, image registration between 2D projections and a 3D image that achieves high accuracy and near real-time computation is challenging. In this paper, we propose a novel method that can rapidly detect an object’s 3D rigid motion or deformation from a 2D projection image or a small set thereof. The method is called CLARET (Correction via Limited-Angle Residues in External Beam Therapy) and consists of two stages: registration preceded by shape space and regression learning. In the registration stage, linear operators are used to iteratively estimate the motion/deformation parameters based on the current intensity residue between the target projec-tion(s) and the digitally reconstructed radiograph(s) (DRRs) of the estimated 3D image. The method determines the linear operators via a two-step learning process. First, it builds a low-order parametric model of the image region’s motion/deformation shape space from its prior 3D images. Second, using learning-time samples produced from the 3D images, it formulates the relationships between the model parameters and the co-varying 2D projection intensity residues by multi-scale linear regressions. The calculated multi-scale regression matrices yield the coarse-to-fine linear operators used in estimating the model parameters from the 2D projection intensity residues in the registration. The method’s application to Image-guided Radiation Therapy (IGRT) requires only a few seconds and yields good results in localizing a tumor under rigid motion in the head and neck and under respiratory deformation in the lung, using one treatment-time imaging 2D projection or a small set thereof. PMID:24058278

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2014-12-02

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

  8. Photothermal imaging scanning microscopy

    DOEpatents

    Chinn, Diane; Stolz, Christopher J.; Wu, Zhouling; Huber, Robert; Weinzapfel, Carolyn

    2006-07-11

    Photothermal Imaging Scanning Microscopy produces a rapid, thermal-based, non-destructive characterization apparatus. Also, a photothermal characterization method of surface and subsurface features includes micron and nanoscale spatial resolution of meter-sized optical materials.

  9. 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.

  10. 3D quantitative analysis of brain SPECT images

    NASA Astrophysics Data System (ADS)

    Loncaric, Sven; Ceskovic, Ivan; Petrovic, Ratimir; Loncaric, Srecko

    2001-07-01

    The main purpose of this work is to develop a computer-based technique for quantitative analysis of 3-D brain images obtained by single photon emission computed tomography (SPECT). In particular, the volume and location of ischemic lesion and penumbra is important for early diagnosis and treatment of infracted regions of the brain. SPECT imaging is typically used as diagnostic tool to assess the size and location of the ischemic lesion. The segmentation method presented in this paper utilizes a 3-D deformable model in order to determine size and location of the regions of interest. The evolution of the model is computed using a level-set implementation of the algorithm. In addition to 3-D deformable model the method utilizes edge detection and region growing for realization of a pre-processing. Initial experimental results have shown that the method is useful for SPECT image analysis.

  11. 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.

  12. Episcopic 3D Imaging Methods: Tools for Researching Gene Function

    PubMed Central

    Weninger, Wolfgang J; Geyer, Stefan H

    2008-01-01

    This work aims at describing episcopic 3D imaging methods and at discussing how these methods can contribute to researching the genetic mechanisms driving embryogenesis and tissue remodelling, and the genesis of pathologies. Several episcopic 3D imaging methods exist. The most advanced are capable of generating high-resolution volume data (voxel sizes from 0.5x0.5x1 µm upwards) of small to large embryos of model organisms and tissue samples. Beside anatomy and tissue architecture, gene expression and gene product patterns can be three dimensionally analyzed in their precise anatomical and histological context with the aid of whole mount in situ hybridization or whole mount immunohistochemical staining techniques. Episcopic 3D imaging techniques were and are employed for analyzing the precise morphological phenotype of experimentally malformed, randomly produced, or genetically engineered embryos of biomedical model organisms. It has been shown that episcopic 3D imaging also fits for describing the spatial distribution of genes and gene products during embryogenesis, and that it can be used for analyzing tissue samples of adult model animals and humans. The latter offers the possibility to use episcopic 3D imaging techniques for researching the causality and treatment of pathologies or for staging cancer. Such applications, however, are not yet routine and currently only preliminary results are available. We conclude that, although episcopic 3D imaging is in its very beginnings, it represents an upcoming methodology, which in short terms will become an indispensable tool for researching the genetic regulation of embryo development as well as the genesis of malformations and diseases. PMID:19452045

  13. Proposed traceable structural resolution protocols for 3D imaging systems

    NASA Astrophysics Data System (ADS)

    MacKinnon, David; Beraldin, J.-Angelo; Cournoyer, Luc; Carrier, Benjamin; Blais, François

    2009-08-01

    A protocol for determining structural resolution using a potentially-traceable reference material is proposed. Where possible, terminology was selected to conform to those published in ISO JCGM 200:2008 (VIM) and ASTM E 2544-08 documents. The concepts of resolvability and edge width are introduced to more completely describe the ability of an optical non-contact 3D imaging system to resolve small features. A distinction is made between 3D range cameras, that obtain spatial data from the total field of view at once, and 3D range scanners, that accumulate spatial data for the total field of view over time. The protocol is presented through the evaluation of a 3D laser line range scanner.

  14. 3D tracking the Brownian motion of colloidal particles using digital holographic microscopy and joint reconstruction.

    PubMed

    Verrier, Nicolas; Fournier, Corinne; Fournel, Thierry

    2015-06-01

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

  15. 3D Analysis of Porosity in a Ceramic Coating Using X-ray Microscopy

    NASA Astrophysics Data System (ADS)

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

    2017-02-01

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

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

    PubMed

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

    2010-01-01

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

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

    PubMed

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

    2010-03-01

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

  18. Image quality enhancement and computation acceleration of 3D holographic display using a symmetrical 3D GS algorithm.

    PubMed

    Zhou, Pengcheng; Bi, Yong; Sun, Minyuan; Wang, Hao; Li, Fang; Qi, Yan

    2014-09-20

    The 3D Gerchberg-Saxton (GS) algorithm can be used to compute a computer-generated hologram (CGH) to produce a 3D holographic display. But, using the 3D GS method, there exists a serious distortion in reconstructions of binary input images. We have eliminated the distortion and improved the image quality of the reconstructions by a maximum of 486%, using a symmetrical 3D GS algorithm that is developed based on a traditional 3D GS algorithm. In addition, the hologram computation speed has been accelerated by 9.28 times, which is significant for real-time holographic displays.

  19. 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.

  20. Efficiency analysis for 3D filtering of multichannel images

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

    Modern remote sensing systems basically acquire images that are multichannel (dual- or multi-polarization, multi- and hyperspectral) where noise, usually with different characteristics, is present in all components. If noise is intensive, it is desirable to remove (suppress) it before applying methods of image classification, interpreting, and information extraction. This can be done using one of two approaches - by component-wise or by vectorial (3D) filtering. The second approach has shown itself to have higher efficiency if there is essential correlation between multichannel image components as this often happens for multichannel remote sensing data of different origin. Within the class of 3D filtering techniques, there are many possibilities and variations. In this paper, we consider filtering based on discrete cosine transform (DCT) and pay attention to two aspects of processing. First, we study in detail what changes in DCT coefficient statistics take place for 3D denoising compared to component-wise processing. Second, we analyze how selection of component images united into 3D data array influences efficiency of filtering and can the observed tendencies be exploited in processing of images with rather large number of channels.

  1. 3D EFT imaging with planar electrode array: Numerical simulation

    NASA Astrophysics Data System (ADS)

    Tuykin, T.; Korjenevsky, A.

    2010-04-01

    Electric field tomography (EFT) is the new modality of the quasistatic electromagnetic sounding of conductive media recently investigated theoretically and realized experimentally. The demonstrated results pertain to 2D imaging with circular or linear arrays of electrodes (and the linear array provides quite poor quality of imaging). In many applications 3D imaging is essential or can increase value of the investigation significantly. In this report we present the first results of numerical simulation of the EFT imaging system with planar array of electrodes which allows 3D visualization of the subsurface conductivity distribution. The geometry of the system is similar to the geometry of our EIT breast imaging system providing 3D conductivity imaging in form of cross-sections set with different depth from the surface. The EFT principle of operation and reconstruction approach differs from the EIT system significantly. So the results of numerical simulation are important to estimate if comparable quality of imaging is possible with the new contactless method. The EFT forward problem is solved using finite difference time domain (FDTD) method for the 8×8 square electrodes array. The calculated results of measurements are used then to reconstruct conductivity distributions by the filtered backprojections along electric field lines. The reconstructed images of the simple test objects are presented.

  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. Reconstruction of 3d Digital Image of Weepingforsythia Pollen

    NASA Astrophysics Data System (ADS)

    Liu, Dongwu; Chen, Zhiwei; Xu, Hongzhi; Liu, Wenqi; Wang, Lina

    Confocal microscopy, which is a major advance upon normal light microscopy, has been used in a number of scientific fields. By confocal microscopy techniques, cells and tissues can be visualized deeply, and three-dimensional images created. Compared with conventional microscopes, confocal microscope improves the resolution of images by eliminating out-of-focus light. Moreover, confocal microscope has a higher level of sensitivity due to highly sensitive light detectors and the ability to accumulate images captured over time. In present studies, a series of Weeping Forsythia pollen digital images (35 images in total) were acquired with confocal microscope, and the three-dimensional digital image of the pollen reconstructed with confocal microscope. Our results indicate that it's a very easy job to analysis threedimensional digital image of the pollen with confocal microscope and the probe Acridine orange (AO).

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

    PubMed Central

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

    2017-01-01

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

  5. 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.

  6. 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

  7. 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

    2016-06-13

    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.

  8. 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.

  9. 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.

  10. 3D FaceCam: a fast and accurate 3D facial imaging device for biometrics applications

    NASA Astrophysics Data System (ADS)

    Geng, Jason; Zhuang, Ping; May, Patrick; Yi, Steven; Tunnell, David

    2004-08-01

    Human faces are fundamentally three-dimensional (3D) objects, and each face has its unique 3D geometric profile. The 3D geometric features of a human face can be used, together with its 2D texture, for rapid and accurate face recognition purposes. Due to the lack of low-cost and robust 3D sensors and effective 3D facial recognition (FR) algorithms, almost all existing FR systems use 2D face images. Genex has developed 3D solutions that overcome the inherent problems in 2D while also addressing limitations in other 3D alternatives. One important aspect of our solution is a unique 3D camera (the 3D FaceCam) that combines multiple imaging sensors within a single compact device to provide instantaneous, ear-to-ear coverage of a human face. This 3D camera uses three high-resolution CCD sensors and a color encoded pattern projection system. The RGB color information from each pixel is used to compute the range data and generate an accurate 3D surface map. The imaging system uses no moving parts and combines multiple 3D views to provide detailed and complete 3D coverage of the entire face. Images are captured within a fraction of a second and full-frame 3D data is produced within a few seconds. This described method provides much better data coverage and accuracy in feature areas with sharp features or details (such as the nose and eyes). Using this 3D data, we have been able to demonstrate that a 3D approach can significantly improve the performance of facial recognition. We have conducted tests in which we have varied the lighting conditions and angle of image acquisition in the "field." These tests have shown that the matching results are significantly improved when enrolling a 3D image rather than a single 2D image. With its 3D solutions, Genex is working toward unlocking the promise of powerful 3D FR and transferring FR from a lab technology into a real-world biometric solution.

  11. Integration of real-time 3D image acquisition and multiview 3D display

    NASA Astrophysics Data System (ADS)

    Zhang, Zhaoxing; Geng, Zheng; Li, Tuotuo; Li, Wei; Wang, Jingyi; Liu, Yongchun

    2014-03-01

    Seamless integration of 3D acquisition and 3D display systems offers enhanced experience in 3D visualization of the real world objects or scenes. The vivid representation of captured 3D objects displayed on a glasses-free 3D display screen could bring the realistic viewing experience to viewers as if they are viewing real-world scene. Although the technologies in 3D acquisition and 3D display have advanced rapidly in recent years, effort is lacking in studying the seamless integration of these two different aspects of 3D technologies. In this paper, we describe our recent progress on integrating a light-field 3D acquisition system and an autostereoscopic multiview 3D display for real-time light field capture and display. This paper focuses on both the architecture design and the implementation of the hardware and the software of this integrated 3D system. A prototype of the integrated 3D system is built to demonstrate the real-time 3D acquisition and 3D display capability of our proposed system.

  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. 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.

  14. 3-D object-oriented image analysis of geophysical data

    NASA Astrophysics Data System (ADS)

    Fadel, I.; Kerle, N.; van der Meijde, M.

    2014-07-01

    Geophysical data are the main source of information about the subsurface. Geophysical techniques are, however, highly non-unique in determining specific physical parameters and boundaries of subsurface objects. To obtain actual physical information, an inversion process is often applied, in which measurements at or above the Earth surface are inverted into a 2- or 3-D subsurface spatial distribution of the physical property. Interpreting these models into structural objects, related to physical processes, requires a priori knowledge and expert analysis which is susceptible to subjective choices and is therefore often non-repeatable. In this research, we implemented a recently introduced object-based approach to interpret the 3-D inversion results of a single geophysical technique using the available a priori information and the physical and geometrical characteristics of the interpreted objects. The introduced methodology is semi-automatic and repeatable, and allows the extraction of subsurface structures using 3-D object-oriented image analysis (3-D OOA) in an objective knowledge-based classification scheme. The approach allows for a semi-objective setting of thresholds that can be tested and, if necessary, changed in a very fast and efficient way. These changes require only changing the thresholds used in a so-called ruleset, which is composed of algorithms that extract objects from a 3-D data cube. The approach is tested on a synthetic model, which is based on a priori knowledge on objects present in the study area (Tanzania). Object characteristics and thresholds were well defined in a 3-D histogram of velocity versus depth, and objects were fully retrieved. The real model results showed how 3-D OOA can deal with realistic 3-D subsurface conditions in which the boundaries become fuzzy, the object extensions become unclear and the model characteristics vary with depth due to the different physical conditions. As expected, the 3-D histogram of the real data was

  15. 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.

  16. Noninvasive computational imaging of cardiac electrophysiology for 3-D infarct.

    PubMed

    Wang, Linwei; Wong, Ken C L; Zhang, Heye; Liu, Huafeng; Shi, Pengcheng

    2011-04-01

    Myocardial infarction (MI) creates electrophysiologically altered substrates that are responsible for ventricular arrhythmias, such as tachycardia and fibrillation. The presence, size, location, and composition of infarct scar bear significant prognostic and therapeutic implications for individual subjects. We have developed a statistical physiological model-constrained framework that uses noninvasive body-surface-potential data and tomographic images to estimate subject-specific transmembrane-potential (TMP) dynamics inside the 3-D myocardium. In this paper, we adapt this framework for the purpose of noninvasive imaging, detection, and quantification of 3-D scar mass for postMI patients: the framework requires no prior knowledge of MI and converges to final subject-specific TMP estimates after several passes of estimation with intermediate feedback; based on the primary features of the estimated spatiotemporal TMP dynamics, we provide 3-D imaging of scar tissue and quantitative evaluation of scar location and extent. Phantom experiments were performed on a computational model of realistic heart-torso geometry, considering 87 transmural infarct scars of different sizes and locations inside the myocardium, and 12 compact infarct scars (extent between 10% and 30%) at different transmural depths. Real-data experiments were carried out on BSP and magnetic resonance imaging (MRI) data from four postMI patients, validated by gold standards and existing results. This framework shows unique advantage of noninvasive, quantitative, computational imaging of subject-specific TMP dynamics and infarct mass of the 3-D myocardium, with the potential to reflect details in the spatial structure and tissue composition/heterogeneity of 3-D infarct scar.

  17. 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

  18. 3D Imaging of Density Gradients Using Plenoptic BOS

    NASA Astrophysics Data System (ADS)

    Klemkowsky, Jenna; Clifford, Chris; Fahringer, Timothy; Thurow, Brian

    2016-11-01

    The combination of background oriented schlieren (BOS) and a plenoptic camera, termed Plenoptic BOS, is explored through two proof-of-concept experiments. The motivation of this work is to provide a 3D technique capable of observing density disturbances. BOS uses the relationship between density and refractive index gradients to observe an apparent shift in a patterned background through image comparison. Conventional BOS systems acquire a single line-of-sight measurement, and require complex configurations to obtain 3D measurements, which are not always conducive to experimental facilities. Plenoptic BOS exploits the plenoptic camera's ability to generate multiple perspective views and refocused images from a single raw plenoptic image during post processing. Using such capabilities, with regards to BOS, provides multiple line-of-sight measurements of density disturbances, which can be collectively used to generate refocused BOS images. Such refocused images allow the position of density disturbances to be qualitatively and quantitatively determined. The image that provides the sharpest density gradient signature corresponds to a specific depth. These results offer motivation to advance Plenoptic BOS with an ultimate goal of reconstructing a 3D density field.

  19. 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.

  20. 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.

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

    PubMed Central

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

    2016-01-01

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

  4. 3D Time-lapse Imaging and Quantification of Mitochondrial Dynamics

    PubMed Central

    Sison, Miguel; Chakrabortty, Sabyasachi; Extermann, Jérôme; Nahas, Amir; James Marchand, Paul; Lopez, Antonio; Weil, Tanja; Lasser, Theo

    2017-01-01

    We present a 3D time-lapse imaging method for monitoring mitochondrial dynamics in living HeLa cells based on photothermal optical coherence microscopy and using novel surface functionalization of gold nanoparticles. The biocompatible protein-based biopolymer coating contains multiple functional groups which impart better cellular uptake and mitochondria targeting efficiency. The high stability of the gold nanoparticles allows continuous imaging over an extended time up to 3000 seconds without significant cell damage. By combining temporal autocorrelation analysis with a classical diffusion model, we quantify mitochondrial dynamics and cast these results into 3D maps showing the heterogeneity of diffusion parameters across the whole cell volume. PMID:28230188

  5. 3D Time-lapse Imaging and Quantification of Mitochondrial Dynamics

    NASA Astrophysics Data System (ADS)

    Sison, Miguel; Chakrabortty, Sabyasachi; Extermann, Jérôme; Nahas, Amir; James Marchand, Paul; Lopez, Antonio; Weil, Tanja; Lasser, Theo

    2017-02-01

    We present a 3D time-lapse imaging method for monitoring mitochondrial dynamics in living HeLa cells based on photothermal optical coherence microscopy and using novel surface functionalization of gold nanoparticles. The biocompatible protein-based biopolymer coating contains multiple functional groups which impart better cellular uptake and mitochondria targeting efficiency. The high stability of the gold nanoparticles allows continuous imaging over an extended time up to 3000 seconds without significant cell damage. By combining temporal autocorrelation analysis with a classical diffusion model, we quantify mitochondrial dynamics and cast these results into 3D maps showing the heterogeneity of diffusion parameters across the whole cell volume.

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. Automated reconstruction of 3D scenes from sequences of images

    NASA Astrophysics Data System (ADS)

    Pollefeys, M.; Koch, R.; Vergauwen, M.; Van Gool, L.

    Modelling of 3D objects from image sequences is a challenging problem and has been an important research topic in the areas of photogrammetry and computer vision for many years. In this paper, a system is presented which automatically extracts a textured 3D surface model from a sequence of images of a scene. The system can deal with unknown camera settings. In addition, the parameters of this camera are allowed to change during acquisition (e.g., by zooming or focusing). No prior knowledge about the scene is necessary to build the 3D models. Therefore, this system offers a high degree of flexibility. The system is based on state-of-the-art algorithms recently developed in computer vision. The 3D modelling task is decomposed into a number of successive steps. Gradually, more knowledge of the scene and the camera setup is retrieved. At this point, the obtained accuracy is not yet at the level required for most metrology applications, but the visual quality is very convincing. This system has been applied to a number of applications in archaeology. The Roman site of Sagalassos (southwest Turkey) was used as a test case to illustrate the potential of this new approach.

  14. 3D imaging of the mesospheric emissive layer

    NASA Astrophysics Data System (ADS)

    Nadjib Kouahla, Mohamed; Faivre, Michael; Moreels, Guy; Clairemidi, Jacques; Mougin-Sisini, Davy; Meriwether, John W.; Lehmacher, Gerald A.; Vidal, Erick; Veliz, Oskar

    A new and original stereo-imaging method is introduced to measure the altitude of the OH airglow layer and provide a 3D map of the altitude of the layer centroid. Near-IR photographs of the layer are taken at two sites distant of 645 km. Each photograph is processed in order to invert the perspective effect and provide a satellite-type 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 crosscorrelation coefficient. 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 2330 m) close to Arequipa. 3D maps of the layer surface are retrieved. They are compared with pseudo-relief intensity maps of the same region. The mean altitude of the emission barycenter is located at 87.1 km on July 26 and 89.5 km on July 28. Comparable relief wavy features appear in the 3D and intensity maps.

  15. Combined registration of 3D tibia and femur implant models in 3D magnetic resonance images

    NASA Astrophysics Data System (ADS)

    Englmeier, Karl-Hans; Siebert, Markus; von Eisenhart-Rothe, Ruediger; Graichen, Heiko

    2008-03-01

    The most frequent reasons for revision of total knee arthroplasty are loosening and abnormal axial alignment leading to an unphysiological kinematic of the knee implant. To get an idea about the postoperative kinematic of the implant, it is essential to determine the position and orientation of the tibial and femoral prosthesis. Therefore we developed a registration method for fitting 3D CAD-models of knee joint prostheses into an 3D MR image. This rigid registration is the basis for a quantitative analysis of the kinematics of knee implants. Firstly the surface data of the prostheses models are converted into a voxel representation; a recursive algorithm determines all boundary voxels of the original triangular surface data. Secondly an initial preconfiguration of the implants by the user is still necessary for the following step: The user has to perform a rough preconfiguration of both remaining prostheses models, so that the fine matching process gets a reasonable starting point. After that an automated gradient-based fine matching process determines the best absolute position and orientation: This iterative process changes all 6 parameters (3 rotational- and 3 translational parameters) of a model by a minimal amount until a maximum value of the matching function is reached. To examine the spread of the final solutions of the registration, the interobserver variability was measured in a group of testers. This variability, calculated by the relative standard deviation, improved from about 50% (pure manual registration) to 0.5% (rough manual preconfiguration and subsequent fine registration with the automatic fine matching process).

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. Image Appraisal for 2D and 3D Electromagnetic Inversion

    SciTech Connect

    Alumbaugh, D.L.; Newman, G.A.

    1999-01-28

    Linearized methods are presented for appraising image resolution and parameter accuracy in images generated with two and three dimensional non-linear electromagnetic inversion schemes. When direct matrix inversion is employed, the model resolution and posterior model covariance matrices can be directly calculated. A method to examine how the horizontal and vertical resolution varies spatially within the electromagnetic property image is developed by examining the columns of the model resolution matrix. Plotting the square root of the diagonal of the model covariance matrix yields an estimate of how errors in the inversion process such as data noise and incorrect a priori assumptions about the imaged model map into parameter error. This type of image is shown to be useful in analyzing spatial variations in the image sensitivity to the data. A method is analyzed for statistically estimating the model covariance matrix when the conjugate gradient method is employed rather than a direct inversion technique (for example in 3D inversion). A method for calculating individual columns of the model resolution matrix using the conjugate gradient method is also developed. Examples of the image analysis techniques are provided on 2D and 3D synthetic cross well EM data sets, as well as a field data set collected at the Lost Hills Oil Field in Central California.

  1. Validation of 3D ultrasound: CT registration of prostate images

    NASA Astrophysics Data System (ADS)

    Firle, Evelyn A.; Wesarg, Stefan; Karangelis, Grigoris; Dold, Christian

    2003-05-01

    All over the world 20% of men are expected to develop prostate cancer sometime in his life. In addition to surgery - being the traditional treatment for cancer - the radiation treatment is getting more popular. The most interesting radiation treatment regarding prostate cancer is Brachytherapy radiation procedure. For the safe delivery of that therapy imaging is critically important. In several cases where a CT device is available a combination of the information provided by CT and 3D Ultrasound (U/S) images offers advantages in recognizing the borders of the lesion and delineating the region of treatment. For these applications the CT and U/S scans should be registered and fused in a multi-modal dataset. Purpose of the present development is a registration tool (registration, fusion and validation) for available CT volumes with 3D U/S images of the same anatomical region, i.e. the prostate. The combination of these two imaging modalities interlinks the advantages of the high-resolution CT imaging and low cost real-time U/S imaging and offers a multi-modality imaging environment for further target and anatomy delineation. This tool has been integrated into the visualization software "InViVo" which has been developed over several years in Fraunhofer IGD in Darmstadt.

  2. 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

  3. 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.

  4. 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

  5. 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.

  6. Imaging and 3D reconstruction of cerebrovascular structures in embryonic zebrafish.

    PubMed

    Ethell, Douglas W; Cameron, D Joshua

    2014-04-22

    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.

  7. Joint calibration of 3D resist image and CDSEM

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

    Traditionally, an optical proximity correction model is to evaluate the resist image at a specific depth within the photoresist and then extract the resist contours from the image. Calibration is generally implemented by comparing resist contours with the critical dimensions (CD). The wafer CD is usually collected by a scanning electron microscope (SEM), which evaluates the CD based on some criterion that is a function of gray level, differential signal, threshold or other parameters set by the SEM. However, the criterion does not reveal which depth the CD is obtained at. This depth inconsistency between modeling and SEM makes the model calibration difficult for low k1 images. In this paper, the vertical resist profile is obtained by modifying the model from planar (2D) to quasi-3D approach and comparing the CD from this new model with SEM CD. For this quasi-3D model, the photoresist diffusion along the depth of the resist is considered and the 3D photoresist contours are evaluated. The performance of this new model is studied and is better than the 2D model.

  8. Digital acquisition system for high-speed 3-D imaging

    NASA Astrophysics Data System (ADS)

    Yafuso, Eiji

    1997-11-01

    High-speed digital three-dimensional (3-D) imagery is possible using multiple independent charge-coupled device (CCD) cameras with sequentially triggered acquisition and individual field storage capability. The system described here utilizes sixteen independent cameras, providing versatility in configuration and image acquisition. By aligning the cameras in nearly coincident lines-of-sight, a sixteen frame two-dimensional (2-D) sequence can be captured. The delays can be individually adjusted lo yield a greater number of acquired frames during the more rapid segments of the event. Additionally, individual integration periods may be adjusted to ensure adequate radiometric response while minimizing image blur. An alternative alignment and triggering scheme arranges the cameras into two angularly separated banks of eight cameras each. By simultaneously triggering correlated stereo pairs, an eight-frame sequence of stereo images may be captured. In the first alignment scheme the camera lines-of-sight cannot be made precisely coincident. Thus representation of the data as a monocular sequence introduces the issue of independent camera coordinate registration with the real scene. This issue arises more significantly using the stereo pair method to reconstruct quantitative 3-D spatial information of the event as a function of time. The principal development here will be the derivation and evaluation of a solution transform and its inverse for the digital data which will yield a 3-D spatial mapping as a function of time.

  9. 3D tongue motion from tagged and cine MR images.

    PubMed

    Xing, Fangxu; Woo, Jonghye; Murano, Emi Z; Lee, Junghoon; Stone, Maureen; Prince, Jerry L

    2013-01-01

    Understanding the deformation of the tongue during human speech is important for head and neck surgeons and speech and language scientists. Tagged magnetic resonance (MR) imaging can be used to image 2D motion, and data from multiple image planes can be combined via post-processing to yield estimates of 3D motion. However, lacking boundary information, this approach suffers from inaccurate estimates near the tongue surface. This paper describes a method that combines two sources of information to yield improved estimation of 3D tongue motion. The method uses the harmonic phase (HARP) algorithm to extract motion from tags and diffeomorphic demons to provide surface deformation. It then uses an incompressible deformation estimation algorithm to incorporate both sources of displacement information to form an estimate of the 3D whole tongue motion. Experimental results show that use of combined information improves motion estimation near the tongue surface, a problem that has previously been reported as problematic in HARP analysis, while preserving accurate internal motion estimates. Results on both normal and abnormal tongue motions are shown.

  10. 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.

  11. Automated spatial alignment of 3D torso images.

    PubMed

    Bose, Arijit; Shah, Shishir K; Reece, Gregory P; Crosby, Melissa A; Beahm, Elisabeth K; Fingeret, Michelle C; Markey, Mia K; Merchant, Fatima A

    2011-01-01

    This paper describes an algorithm for automated spatial alignment of three-dimensional (3D) surface images in order to achieve a pre-defined orientation. Surface images of the torso are acquired from breast cancer patients undergoing reconstructive surgery to facilitate objective evaluation of breast morphology pre-operatively (for treatment planning) and/or post-operatively (for outcome assessment). Based on the viewing angle of the multiple cameras used for stereophotography, the orientation of the acquired torso in the images may vary from the normal upright position. Consequently, when translating this data into a standard 3D framework for visualization and analysis, the co-ordinate geometry differs from the upright position making robust and standardized comparison of images impractical. Moreover, manual manipulation and navigation of images to the desired upright position is subject to user bias. Automating the process of alignment and orientation removes operator bias and permits robust and repeatable adjustment of surface images to a pre-defined or desired spatial geometry.

  12. 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.

  13. Integral imaging based 3D display of holographic data.

    PubMed

    Yöntem, Ali Özgür; Onural, Levent

    2012-10-22

    We propose a method and present applications of this method that converts a diffraction pattern into an elemental image set in order to display them on an integral imaging based display setup. We generate elemental images based on diffraction calculations as an alternative to commonly used ray tracing methods. Ray tracing methods do not accommodate the interference and diffraction phenomena. Our proposed method enables us to obtain elemental images from a holographic recording of a 3D object/scene. The diffraction pattern can be either numerically generated data or digitally acquired optical data. The method shows the connection between a hologram (diffraction pattern) and an elemental image set of the same 3D object. We showed three examples, one of which is the digitally captured optical diffraction tomography data of an epithelium cell. We obtained optical reconstructions with our integral imaging display setup where we used a digital lenslet array. We also obtained numerical reconstructions, again by using the diffraction calculations, for comparison. The digital and optical reconstruction results are in good agreement.

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

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

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

  15. A hybrid framework for 3D medical image segmentation.

    PubMed

    Chen, Ting; Metaxas, Dimitris

    2005-12-01

    In this paper we propose a novel hybrid 3D segmentation framework which combines Gibbs models, marching cubes and deformable models. In the framework, first we construct a new Gibbs model whose energy function is defined on a high order clique system. The new model includes both region and boundary information during segmentation. Next we improve the original marching cubes method to construct 3D meshes from Gibbs models' output. The 3D mesh serves as the initial geometry of the deformable model. Then we deform the deformable model using external image forces so that the model converges to the object surface. We run the Gibbs model and the deformable model recursively by updating the Gibbs model's parameters using the region and boundary information in the deformable model segmentation result. In our approach, the hybrid combination of region-based methods and boundary-based methods results in improved segmentations of complex structures. The benefit of the methodology is that it produces high quality segmentations of 3D structures using little prior information and minimal user intervention. The modules in this segmentation methodology are developed within the context of the Insight ToolKit (ITK). We present experimental segmentation results of brain tumors and evaluate our method by comparing experimental results with expert manual segmentations. The evaluation results show that the methodology achieves high quality segmentation results with computational efficiency. We also present segmentation results of other clinical objects to illustrate the strength of the methodology as a generic segmentation framework.

  16. 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.

  17. 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.

  18. Virtual image display as a backlight for 3D.

    PubMed

    Travis, Adrian; MacCrann, Niall; Emerton, Neil; Kollin, Joel; Georgiou, Andreas; Lanier, Jaron; Bathiche, Stephen

    2013-07-29

    We describe a device which has the potential to be used both as a virtual image display and as a backlight. The pupil of the emitted light fills the device approximately to its periphery and the collimated emission can be scanned both horizontally and vertically in the manner needed to illuminate an eye in any position. The aim is to reduce the power needed to illuminate a liquid crystal panel but also to enable a smooth transition from 3D to a virtual image as the user nears the screen.

  19. 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.

  20. 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

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

    PubMed

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

    2015-07-10

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

  4. 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.

  5. Quantification of thyroid volume using 3-D ultrasound imaging.

    PubMed

    Kollorz, E K; Hahn, D A; Linke, R; Goecke, T W; Hornegger, J; Kuwert, T

    2008-04-01

    Ultrasound (US) is among the most popular diagnostic techniques today. It is non-invasive, fast, comparably cheap, and does not require ionizing radiation. US is commonly used to examine the size, and structure of the thyroid gland. In clinical routine, thyroid imaging is usually performed by means of 2-D US. Conventional approaches for measuring the volume of the thyroid gland or its nodules may therefore be inaccurate due to the lack of 3-D information. This work reports a semi-automatic segmentation approach for the classification, and analysis of the thyroid gland based on 3-D US data. The images are scanned in 3-D, pre-processed, and segmented. Several pre-processing methods, and an extension of a commonly used geodesic active contour level set formulation are discussed in detail. The results obtained by this approach are compared to manual interactive segmentations by a medical expert in five representative patients. Our work proposes a novel framework for the volumetric quantification of thyroid gland lobes, which may also be expanded to other parenchymatous organs.

  6. 3D imaging of biological specimen using MS.

    PubMed

    Fletcher, John S

    2015-01-01

    Imaging MS can provide unique information about the distribution of native and non-native compounds in biological specimen. MALDI MS and secondary ion MS are the two most commonly applied imaging MS techniques and can provide complementary information about a sample. MALDI offers access to high mass species such as proteins while secondary ion MS can operate at higher spatial resolution and provide information about lower mass species including elemental signals. Imaging MS is not limited to two dimensions and different approaches have been developed that allow 3D molecular images to be generated of chemicals in whole organs down to single cells. Resolution in the z-dimension is often higher than in x and y, so such analysis offers the potential for probing the distribution of drug molecules and studying drug action by MS with a much higher precision - possibly even organelle level.

  7. 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.

  8. 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

  9. 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.

  10. Evaluation of Kinect 3D Sensor for Healthcare Imaging.

    PubMed

    Pöhlmann, Stefanie T L; Harkness, Elaine F; Taylor, Christopher J; Astley, Susan M

    2016-01-01

    Microsoft Kinect is a three-dimensional (3D) sensor originally designed for gaming that has received growing interest as a cost-effective and safe device for healthcare imaging. Recent applications of Kinect in health monitoring, screening, rehabilitation, assistance systems, and intervention support are reviewed here. The suitability of available technologies for healthcare imaging applications is assessed. The performance of Kinect I, based on structured light technology, is compared with that of the more recent Kinect II, which uses time-of-flight measurement, under conditions relevant to healthcare applications. The accuracy, precision, and resolution of 3D images generated with Kinect I and Kinect II are evaluated using flat cardboard models representing different skin colors (pale, medium, and dark) at distances ranging from 0.5 to 1.2 m and measurement angles of up to 75°. Both sensors demonstrated high accuracy (majority of measurements <2 mm) and precision (mean point to plane error <2 mm) at an average resolution of at least 390 points per cm(2). Kinect I is capable of imaging at shorter measurement distances, but Kinect II enables structures angled at over 60° to be evaluated. Kinect II showed significantly higher precision and Kinect I showed significantly higher resolution (both p < 0.001). The choice of object color can influence measurement range and precision. Although Kinect is not a medical imaging device, both sensor generations show performance adequate for a range of healthcare imaging applications. Kinect I is more appropriate for short-range imaging and Kinect II is more appropriate for imaging highly curved surfaces such as the face or breast.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

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

  16. 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

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

    PubMed

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

    2013-08-12

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

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

    PubMed

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

    2017-01-01

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

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

    DTIC Science & Technology

    2014-05-01

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

  20. Image segmentation to inspect 3-D object sizes

    NASA Astrophysics Data System (ADS)

    Hsu, Jui-Pin; Fuh, Chiou-Shann

    1996-01-01

    Object size inspection is an important task and has various applications in computer vision. For example, the automatic control of stone-breaking machines, which perform better if the sizes of the stones to be broken can be predicted. An algorithm is proposed for image segmentation in size inspection for almost round stones with high or low texture. Although our experiments are focused on stones, the algorithm can be applied to other 3-D objects. We use one fixed camera and four light sources at four different positions one at a time, to take four images. Then we compute the image differences and binarize them to extract edges. We explain, step by step, the photographing, the edge extraction, the noise removal, and the edge gap filling. Experimental results are presented.

  1. The Diagnostic Radiological Utilization Of 3-D Display Images

    NASA Astrophysics Data System (ADS)

    Cook, Larry T.; Dwyer, Samuel J.; Preston, David F.; Batnitzky, Solomon; Lee, Kyo R.

    1984-10-01

    In the practice of radiology, computer graphics systems have become an integral part of the use of computed tomography (CT), nuclear medicine (NM), magnetic resonance imaging (MRI), digital subtraction angiography (DSA) and ultrasound. Gray scale computerized display systems are used to display, manipulate, and record scans in all of these modalities. As the use of these imaging systems has spread, various applications involving digital image manipulation have also been widely accepted in the radiological community. We discuss one of the more esoteric of such applications, namely, the reconstruction of 3-D structures from plane section data, such as CT scans. Our technique is based on the acquisition of contour data from successive sections, the definition of the implicit surface defined by such contours, and the application of the appropriate computer graphics hardware and software to present reasonably pleasing pictures.

  2. 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.

  3. Low cost 3D scanning process using digital image processing

    NASA Astrophysics Data System (ADS)

    Aguilar, David; Romero, Carlos; Martínez, Fernando

    2017-02-01

    This paper shows the design and building of a low cost 3D scanner, able to digitize solid objects through contactless data acquisition, using active object reflection. 3D scanners are used in different applications such as: science, engineering, entertainment, etc; these are classified in: contact scanners and contactless ones, where the last ones are often the most used but they are expensive. This low-cost prototype is done through a vertical scanning of the object using a fixed camera and a mobile horizontal laser light, which is deformed depending on the 3-dimensional surface of the solid. Using digital image processing an analysis of the deformation detected by the camera was done; it allows determining the 3D coordinates using triangulation. The obtained information is processed by a Matlab script, which gives to the user a point cloud corresponding to each horizontal scanning done. The obtained results show an acceptable quality and significant details of digitalized objects, making this prototype (built on LEGO Mindstorms NXT kit) a versatile and cheap tool, which can be used for many applications, mainly by engineering students.

  4. Effective classification of 3D image data using partitioning methods

    NASA Astrophysics Data System (ADS)

    Megalooikonomou, Vasileios; Pokrajac, Dragoljub; Lazarevic, Aleksandar; Obradovic, Zoran

    2002-03-01

    We propose partitioning-based methods to facilitate the classification of 3-D binary image data sets of regions of interest (ROIs) with highly non-uniform distributions. The first method is based on recursive dynamic partitioning of a 3-D volume into a number of 3-D hyper-rectangles. For each hyper-rectangle, we consider, as a potential attribute, the number of voxels (volume elements) that belong to ROIs. A hyper-rectangle is partitioned only if the corresponding attribute does not have high discriminative power, determined by statistical tests, but it is still sufficiently large for further splitting. The final discriminative hyper-rectangles form new attributes that are further employed in neural network classification models. The second method is based on maximum likelihood employing non-spatial (k-means) and spatial DBSCAN clustering algorithms to estimate the parameters of the underlying distributions. The proposed methods were experimentally evaluated on mixtures of Gaussian distributions, on realistic lesion-deficit data generated by a simulator conforming to a clinical study, and on synthetic fractal data. Both proposed methods have provided good classification on Gaussian mixtures and on realistic data. However, the experimental results on fractal data indicated that the clustering-based methods were only slightly better than random guess, while the recursive partitioning provided significantly better classification accuracy.

  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. 3D imaging reconstruction and impacted third molars: case reports

    PubMed Central

    Tuzi, Andrea; Di Bari, Roberto; Cicconetti, Andrea

    2012-01-01

    Summary There is a debate in the literature about the need for Computed Tomagraphy (CT) before removing third molars, even if positive radiographic signs are present. In few cases, the third molar is so close to the inferior alveolar nerve that its extraction might expose patients to the risk of post-operative neuro-sensitive alterations of the skin and the mucosa of the homolateral lower lip and chin. Thus, the injury of the inferior alveolar nerve may represent a serious, though infrequent, neurologic complication in the surgery of the third molars rendering necessary a careful pre-operative evaluation of their anatomical relationship with the inferior alveolar nerve by means of radiographic imaging techniques. This contribution presents two case reports showing positive radiographic signs, which are the hallmarks of a possible close relationship between the inferior alveolar nerve and the third molars. We aim at better defining the relationship between third molars and the mandibular canal using Dental CT Scan, DICOM image acquisition and 3D reconstruction with a dedicated software. By our study we deduce that 3D images are not indispensable, but they can provide a very agreeable assistance in the most complicated cases. PMID:23386934

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

    PubMed

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

    2017-03-18

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

  8. 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.).

  9. 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.

  10. 3-D Imaging of Partly Concealed Targets by Laser Radar

    DTIC Science & Technology

    2005-10-01

    laser in the green wavelength region was used for illumination. 3-D Imaging of Partly Concealed Targets by Laser Radar 11 - 8 RTO-MP-SET-094...acknowledge Marie Carlsson and Ann Charlotte Gustavsson for their assistance in some of the experiments. 7.0 REFERENCES [1] U. Söderman, S. Ahlberg...SPIE Vol. 3707, pp. 432-448, USA, 1999. [14] D. Letalick, H. Larsson, M. Carlsson, and A.-C. Gustavsson , “Laser sensors for urban warfare,” FOI

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

    PubMed

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

    2013-05-20

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

  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. Finite element 3D reconstruction of the pulmonary acinus imaged by synchrotron X-ray tomography

    PubMed Central

    Tsuda, A.; Filipovic, N.; Haberthür, D.; Dickie, R.; Matsui, Y.; Stampanoni, M.; Schittny, J. C.

    2008-01-01

    The alveolated structure of the pulmonary acinus plays a vital role in gas exchange function. Three-dimensional (3D) analysis of the parenchymal region is fundamental to understanding this structure-function relationship, but only a limited number of attempts have been conducted in the past because of technical limitations. In this study, we developed a new image processing methodology based on finite element (FE) analysis for accurate 3D structural reconstruction of the gas exchange regions of the lung. Stereologically well characterized rat lung samples (Pediatr Res 53: 72–80, 2003) were imaged using high-resolution synchrotron radiation-based X-ray tomographic microscopy. A stack of 1,024 images (each slice: 1024 × 1024 pixels) with resolution of 1.4 μm3 per voxel were generated. For the development of FE algorithm, regions of interest (ROI), containing ∼7.5 million voxels, were further extracted as a working subunit. 3D FEs were created overlaying the voxel map using a grid-based hexahedral algorithm. A proper threshold value for appropriate segmentation was iteratively determined to match the calculated volume density of tissue to the stereologically determined value (Pediatr Res 53: 72–80, 2003). The resulting 3D FEs are ready to be used for 3D structural analysis as well as for subsequent FE computational analyses like fluid dynamics and skeletonization. PMID:18583378

  14. 3D painting documentation: evaluation of conservation conditions with 3D imaging and ranging techniques

    NASA Astrophysics Data System (ADS)

    Abate, D.; Menna, F.; Remondino, F.; Gattari, M. G.

    2014-06-01

    The monitoring of paintings, both on canvas and wooden support, is a crucial issue for the preservation and conservation of this kind of artworks. Many environmental factors (e.g. humidity, temperature, illumination, etc.), as well as bad conservation practices (e.g. wrong restorations, inappropriate locations, etc.), can compromise the material conditions over time and deteriorate an artwork. The article presents an on-going project realized by a multidisciplinary team composed by the ENEA UTICT 3D GraphLab, the 3D Optical Metrology Unit of the Bruno Kessler Foundation and the Soprintendenza per i Beni Storico Artistici ed Etnoantropologici of Bologna (Italy). The goal of the project is the multi-temporal 3D documentation and monitoring of paintings - at the moment in bad conservation's situation - and the provision of some metrics to quantify the deformations and damages.

  15. 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.

  16. Imaging Shallow Salt With 3D Refraction Migration

    NASA Astrophysics Data System (ADS)

    Vanschuyver, C. J.; Hilterman, F. J.

    2005-05-01

    In offshore West Africa, numerous salt walls are within 200 m of sea level. Because of the shallowness of these salt walls, reflections from the salt top can be difficult to map, making it impossible to build an accurate velocity model for subsequent pre-stack depth migration. An accurate definition of salt boundaries is critical to any depth model where salt is present. Unfortunately, when a salt body is very shallow, the reflection from the upper interface can be obscured due to large offsets between the source and near receivers and also due to the interference from multiples and other near-surface noise events. A new method is described using 3D migration of the refraction waveforms which is simplified because of several constraints in the model definition. The azimuth and dip of the refractor is found by imaging with Kirchhoff theory. A Kirchhoff migration is performed where the traveltime values are adjusted to use the CMP refraction traveltime equation. I assume the sediment and salt velocities to be known such that once the image time is specified, then the dip and azimuth of the refraction path can be found. The resulting 3D refraction migrations are in excellent depth agreement with available well control. In addition, the refraction migration time picks of deeper salt events are in agreement with time picks of the same events on the reflection migration.

  17. 3-D visualization and animation technologies in anatomical imaging.

    PubMed

    McGhee, John

    2010-02-01

    This paper explores a 3-D computer artist's approach to the creation of three-dimensional computer-generated imagery (CGI) derived from clinical scan data. Interpretation of scientific imagery, such as magnetic resonance imaging (MRI), is restricted to the eye of the trained medical practitioner in a clinical or scientific context. In the research work described here, MRI data are visualized and interpreted by a 3-D computer artist using the tools of the digital animator to navigate image complexity and widen interaction. In this process, the artefact moves across disciplines; it is no longer tethered to its diagnostic origins. It becomes an object that has visual attributes such as light, texture and composition, and a visual aesthetic of its own. The introduction of these visual attributes provides a platform for improved accessibility by a lay audience. The paper argues that this more artisan approach to clinical data visualization has a potential real-world application as a communicative tool for clinicians and patients during consultation.

  18. 3-D visualization and animation technologies in anatomical imaging

    PubMed Central

    McGhee, John

    2010-01-01

    This paper explores a 3-D computer artist’s approach to the creation of three-dimensional computer-generated imagery (CGI) derived from clinical scan data. Interpretation of scientific imagery, such as magnetic resonance imaging (MRI), is restricted to the eye of the trained medical practitioner in a clinical or scientific context. In the research work described here, MRI data are visualized and interpreted by a 3-D computer artist using the tools of the digital animator to navigate image complexity and widen interaction. In this process, the artefact moves across disciplines; it is no longer tethered to its diagnostic origins. It becomes an object that has visual attributes such as light, texture and composition, and a visual aesthetic of its own. The introduction of these visual attributes provides a platform for improved accessibility by a lay audience. The paper argues that this more artisan approach to clinical data visualization has a potential real-world application as a communicative tool for clinicians and patients during consultation. PMID:20002229

  19. 3-D Imaging and Simulation for Nephron Sparing Surgical Training.

    PubMed

    Ahmadi, Hamed; Liu, Jen-Jane

    2016-08-01

    Minimally invasive partial nephrectomy (MIPN) is now considered the procedure of choice for small renal masses largely based on functional advantages over traditional open surgery. Lack of haptic feedback, the need for spatial understanding of tumor borders, and advanced operative techniques to minimize ischemia time or achieve zero-ischemia PN are among factors that make MIPN a technically demanding operation with a steep learning curve for inexperienced surgeons. Surgical simulation has emerged as a useful training adjunct in residency programs to facilitate the acquisition of these complex operative skills in the setting of restricted work hours and limited operating room time and autonomy. However, the majority of available surgical simulators focus on basic surgical skills, and procedure-specific simulation is needed for optimal surgical training. Advances in 3-dimensional (3-D) imaging have also enhanced the surgeon's ability to localize tumors intraoperatively. This article focuses on recent procedure-specific simulation models for laparoscopic and robotic-assisted PN and advanced 3-D imaging techniques as part of pre- and some cases, intraoperative surgical planning.

  20. Abdominal aortic aneurysm imaging with 3-D ultrasound: 3-D-based maximum diameter measurement and volume quantification.

    PubMed

    Long, A; Rouet, L; Debreuve, A; Ardon, R; Barbe, C; Becquemin, J P; Allaire, E

    2013-08-01

    The clinical reliability of 3-D ultrasound imaging (3-DUS) in quantification of abdominal aortic aneurysm (AAA) was evaluated. B-mode and 3-DUS images of AAAs were acquired for 42 patients. AAAs were segmented. A 3-D-based maximum diameter (Max3-D) and partial volume (Vol30) were defined and quantified. Comparisons between 2-D (Max2-D) and 3-D diameters and between orthogonal acquisitions were performed. Intra- and inter-observer reproducibility was evaluated. Intra- and inter-observer coefficients of repeatability (CRs) were less than 5.18 mm for Max3-D. Intra-observer and inter-observer CRs were respectively less than 6.16 and 8.71 mL for Vol30. The mean of normalized errors of Vol30 was around 7%. Correlation between Max2-D and Max3-D was 0.988 (p < 0.0001). Max3-D and Vol30 were not influenced by a probe rotation of 90°. Use of 3-DUS to quantify AAA is a new approach in clinical practice. The present study proposed and evaluated dedicated parameters. Their reproducibility makes the technique clinically reliable.

  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. 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

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

    PubMed

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

    2014-10-01

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

  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. 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.

  6. Complex Resistivity 3D Imaging for Ground Reinforcement Site

    NASA Astrophysics Data System (ADS)

    Son, J.; Kim, J.; Park, S.

    2012-12-01

    Induced polarization (IP) method is used for mineral exploration and generally classified into two categories, time and frequency domain method. IP method in frequency domain measures amplitude and absolute phase to the transmitted currents, and is often called spectral induced polarization (SIP) when measurement is made for the wide-band frequencies. Our research group has been studying the modeling and inversion algorithms of complex resistivity method since several years ago and recently started to apply this method for various field applications. We already completed the development of 2/3D modeling and inversion program and developing another algorithm to use wide-band data altogether. Until now complex resistivity (CR) method was mainly used for the surface or tomographic survey of mineral exploration. Through the experience, we can find that the resistivity section from CR method is very similar with that of conventional resistivity method. Interpretation of the phase section is generally well matched with the geological information of survey area. But because most of survey area has very touch and complex terrain, 2D survey and interpretation are used generally. In this study, the case study of 3D CR survey conducted for the site where ground reinforcement was done to prevent the subsidence will be introduced. Data was acquired with the Zeta system, the complex resistivity measurement system produced by Zonge Co. using 8 frequencies from 0.125 to 16 Hz. 2D survey was conducted for total 6 lines with 5 m dipole spacing and 20 electrodes. Line length is 95 meter for every line. Among these 8 frequency data, data below 1 Hz was used considering its quality. With the 6 line data, 3D inversion was conducted. Firstly 2D interpretation was made with acquired data and its results were compared with those of resistivity survey. Resulting resistivity image sections of CR and resistivity method were very similar. Anomalies in phase image section showed good agreement

  7. High Time Resolution Photon Counting 3D Imaging Sensors

    NASA Astrophysics Data System (ADS)

    Siegmund, O.; Ertley, C.; Vallerga, J.

    2016-09-01

    Novel sealed tube microchannel plate (MCP) detectors using next generation cross strip (XS) anode readouts and high performance electronics have been developed to provide photon counting imaging sensors for Astronomy and high time resolution 3D remote sensing. 18 mm aperture sealed tubes with MCPs and high efficiency Super-GenII or GaAs photocathodes have been implemented to access the visible/NIR regimes for ground based research, astronomical and space sensing applications. The cross strip anode readouts in combination with PXS-II high speed event processing electronics can process high single photon counting event rates at >5 MHz ( 80 ns dead-time per event), and time stamp events to better than 25 ps. Furthermore, we are developing a high speed ASIC version of the electronics for low power/low mass spaceflight applications. For a GaAs tube the peak quantum efficiency has degraded from 30% (at 560 - 850 nm) to 25% over 4 years, but for Super-GenII tubes the peak quantum efficiency of 17% (peak at 550 nm) has remained unchanged for over 7 years. The Super-GenII tubes have a uniform spatial resolution of <30 μm FWHM ( 1 x106 gain) and single event timing resolution of 100 ps (FWHM). The relatively low MCP gain photon counting operation also permits longer overall sensor lifetimes and high local counting rates. Using the high timing resolution, we have demonstrated 3D object imaging with laser pulse (630 nm 45 ps jitter Pilas laser) reflections in single photon counting mode with spatial and depth sensitivity of the order of a few millimeters. A 50 mm Planacon sealed tube was also constructed, using atomic layer deposited microchannel plates which potentially offer better overall sealed tube lifetime, quantum efficiency and gain stability. This tube achieves standard bialkali quantum efficiency levels, is stable, and has been coupled to the PXS-II electronics and used to detect and image fast laser pulse signals.

  8. 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

  9. 3D texture analysis for classification of second harmonic generation images of human ovarian cancer

    NASA Astrophysics Data System (ADS)

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

    2016-10-01

    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.

  10. 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.

  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. Fast 3-d tomographic microwave imaging for breast cancer detection.

    PubMed

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

    2012-08-01

    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.

  13. 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.

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

    PubMed

    Esquivel-Rodríguez, Juan; Kihara, Daisuke

    2013-10-01

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

  15. 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.

  16. 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.

  17. Image sequence coding using 3D scene models

    NASA Astrophysics Data System (ADS)

    Girod, Bernd

    1994-09-01

    The implicit and explicit use of 3D models for image sequence coding is discussed. For implicit use, a 3D model can be incorporated into motion compensating prediction. A scheme that estimates the displacement vector field with a rigid body motion constraint by recovering epipolar lines from an unconstrained displacement estimate and then repeating block matching along the epipolar line is proposed. Experimental results show that an improved displacement vector field can be obtained with a rigid body motion constraint. As an example for explicit use, various results with a facial animation model for videotelephony are discussed. A 13 X 16 B-spline mask can be adapted automatically to individual faces and is used to generate facial expressions based on FACS. A depth-from-defocus range camera suitable for real-time facial motion tracking is described. Finally, the real-time facial animation system `Traugott' is presented that has been used to generate several hours of broadcast video. Experiments suggest that a videophone system based on facial animation might require a transmission bitrate of 1 kbit/s or below.

  18. 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

  19. Unsupervised fuzzy segmentation of 3D magnetic resonance brain images

    NASA Astrophysics Data System (ADS)

    Velthuizen, Robert P.; Hall, Lawrence O.; Clarke, Laurence P.; Bensaid, Amine M.; Arrington, J. A.; Silbiger, Martin L.

    1993-07-01

    Unsupervised fuzzy methods are proposed for segmentation of 3D Magnetic Resonance images of the brain. Fuzzy c-means (FCM) has shown promising results for segmentation of single slices. FCM has been investigated for volume segmentations, both by combining results of single slices and by segmenting the full volume. Different strategies and initializations have been tried. In particular, two approaches have been used: (1) a method by which, iteratively, the furthest sample is split off to form a new cluster center, and (2) the traditional FCM in which the membership grade matrix is initialized in some way. Results have been compared with volume segmentations by k-means and with two supervised methods, k-nearest neighbors and region growing. Results of individual segmentations are presented as well as comparisons on the application of the different methods to a number of tumor patient data sets.

  20. 3D x-ray reconstruction using lightfield imaging

    NASA Astrophysics Data System (ADS)

    Saha, Sajib; Tahtali, Murat; Lambert, Andrew; Pickering, Mark R.

    2014-09-01

    Existing Computed Tomography (CT) systems require full 360° rotation projections. Using the principles of lightfield imaging, only 4 projections under ideal conditions can be sufficient when the object is illuminated with multiple-point Xray sources. The concept was presented in a previous work with synthetically sampled data from a synthetic phantom. Application to real data requires precise calibration of the physical set up. This current work presents the calibration procedures along with experimental findings for the reconstruction of a physical 3D phantom consisting of simple geometric shapes. The crucial part of this process is to determine the effective distances of the X-ray paths, which are not possible or very difficult by direct measurements. Instead, they are calculated by tracking the positions of fiducial markers under prescribed source and object movements. Iterative algorithms are used for the reconstruction. Customized backprojection is used to ensure better initial guess for the iterative algorithms to start with.

  1. 3D imaging of semiconductor components by discrete laminography

    NASA Astrophysics Data System (ADS)

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

    2014-06-01

    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.

  2. 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.

  3. 3D and multispectral imaging for subcutaneous veins detection.

    PubMed

    Paquit, Vincent C; Tobin, Kenneth W; Price, Jeffery R; Mèriaudeau, Fabrice

    2009-07-06

    The first and perhaps most important phase of a surgical procedure is the insertion of an intravenous (IV) catheter. Currently, this is performed manually by trained personnel. In some visions of future operating rooms, however, this process is to be replaced by an automated system. Experiments to determine the best NIR wavelengths to optimize vein contrast for physiological differences such as skin tone and/or the presence of hair on the arm or wrist surface are presented. For illumination our system is composed of a mercury arc lamp coupled to a 10nm band-pass spectrometer. A structured lighting system is also coupled to our multispectral system in order to provide 3D information of the patient arm orientation. Images of each patient arm are captured under every possible combinations of illuminants and the optimal combination of wavelengths for a given subject to maximize vein contrast using linear discriminant analysis is determined.

  4. 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.

  5. 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

  6. 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.

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

    NASA Astrophysics Data System (ADS)

    Kreis, T.

    2012-03-01

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

  8. 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.

  9. 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

  10. Nonrigid Registration of 2-D and 3-D Dynamic Cell Nuclei Images for Improved Classification of Subcellular Particle Motion

    PubMed Central

    Kim, Il-Han; Chen, Yi-Chun M.; Spector, David L.; Eils, Roland; Rohr, Karl

    2012-01-01

    The observed motion of subcellular particles in fluorescence microscopy image sequences of live cells is generally a superposition of the motion and deformation of the cell and the motion of the particles. Decoupling the two types of movements to enable accurate classification of the particle motion requires the application of registration algorithms. We have developed an intensity-based approach for nonrigid registration of multi-channel microscopy image sequences of cell nuclei. First, based on 3-D synthetic images we demonstrate that cell nucleus deformations change the observed motion types of particles and that our approach allows to recover the original motion. Second, we have successfully applied our approach to register 2-D and 3-D real microscopy image sequences. A quantitative experimental comparison with previous approaches for nonrigid registration of cell microscopy has also been performed. PMID:20840894

  11. 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.

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

    SciTech Connect

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

    2011-01-01

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

  13. 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.

  14. A Simple Quality Assessment Index for Stereoscopic Images Based on 3D Gradient Magnitude

    PubMed Central

    Wang, Shanshan; Shao, Feng; Li, Fucui; Yu, Mei; Jiang, Gangyi

    2014-01-01

    We present a simple quality assessment index for stereoscopic images based on 3D gradient magnitude. To be more specific, we construct 3D volume from the stereoscopic images across different disparity spaces and calculate pointwise 3D gradient magnitude similarity (3D-GMS) along three horizontal, vertical, and viewpoint directions. Then, the quality score is obtained by averaging the 3D-GMS scores of all points in the 3D volume. Experimental results on four publicly available 3D image quality assessment databases demonstrate that, in comparison with the most related existing methods, the devised algorithm achieves high consistency alignment with subjective assessment. PMID:25133265

  15. 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

  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. 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.

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

    NASA Astrophysics Data System (ADS)

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

    2006-03-01

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

  19. 3D Seismic Imaging over a Potential Collapse Structure

    NASA Astrophysics Data System (ADS)

    Gritto, Roland; O'Connell, Daniel; Elobaid Elnaiem, Ali; Mohamed, Fathelrahman; Sadooni, Fadhil

    2016-04-01

    The Middle-East has seen a recent boom in construction including the planning and development of complete new sub-sections of metropolitan areas. Before planning and construction can commence, however, the development areas need to be investigated to determine their suitability for the planned project. Subsurface parameters such as the type of material (soil/rock), thickness of top soil or rock layers, depth and elastic parameters of basement, for example, comprise important information needed before a decision concerning the suitability of the site for construction can be made. A similar problem arises in environmental impact studies, when subsurface parameters are needed to assess the geological heterogeneity of the subsurface. Environmental impact studies are typically required for each construction project, particularly for the scale of the aforementioned building boom in the Middle East. The current study was conducted in Qatar at the location of a future highway interchange to evaluate a suite of 3D seismic techniques in their effectiveness to interrogate the subsurface for the presence of karst-like collapse structures. The survey comprised an area of approximately 10,000 m2 and consisted of 550 source- and 192 receiver locations. The seismic source was an accelerated weight drop while the geophones consisted of 3-component 10 Hz velocity sensors. At present, we analyzed over 100,000 P-wave phase arrivals and performed high-resolution 3-D tomographic imaging of the shallow subsurface. Furthermore, dispersion analysis of recorded surface waves will be performed to obtain S-wave velocity profiles of the subsurface. Both results, in conjunction with density estimates, will be utilized to determine the elastic moduli of the subsurface rock layers.

  20. Venus Topography in 3D: Imaging of Coronae and Chasmata

    NASA Astrophysics Data System (ADS)

    Jurdy, D. M.; Stefanick, M.; Stoddard, P. R.

    2006-12-01

    Venus' surface hosts hundreds of circular to elongate features, ranging from 60-2600 km, and averaging somewhat over 200 km, in diameter. These enigmatic structures have been classified as "coronae" and attributed to either tectono-volcanic or impact-related mechanisms. A linear to arcuate system of chasmata - rugged zones with some of Venus' deepest troughs, extend 1000's of kilometers. They have extreme relief, with elevations changing as much as 7 km in just 30 km distance. The 54,464 km-long Venus chasmata system defined in great detail by Magellan can be fit by great circle arcs at the 89.6% level, and when corrected for the smaller size of the planet, the total length of the chasmata system measures within 2.7% of the length of Earth's spreading ridges. The relatively young Beta-Atla-Themis region (BAT), within 30° of the equator from 180-300° longitude has the planet's strongest geoid highs and profuse volcanism. This BAT region, the intersection of three rift zones, also has a high coronal concentration, with individual coronae closely associated with the chasmata system. The chasmata with the greatest relief on Venus show linear rifting that prevailed in the latest stage of tectonic deformation. For a three-dimensional view of Venus' surface, we spread out the Magellan topography on a flat surface using a Mercator projection to preserve shape. Next we illuminate the surface with beams at angle 45° from left (or right) so as to simulate mid afternoon (or mid-morning). Finally, we observe the surface with two eyes looking through orange and azure colored filters respectively. This gives a 3D view of tectonic features in the BAT area. The 3D images clearly show coronae sharing boundaries with the chasmata. This suggests that the processes of rifting and corona-formation occur together. It seems unlikely that impact craters would create this pattern.

  1. Autostereoscopic 3D visualization and image processing system for neurosurgery.

    PubMed

    Meyer, Tobias; Kuß, Julia; Uhlemann, Falk; Wagner, Stefan; Kirsch, Matthias; Sobottka, Stephan B; Steinmeier, Ralf; Schackert, Gabriele; Morgenstern, Ute

    2013-06-01

    A demonstrator system for planning neurosurgical procedures was developed based on commercial hardware and software. The system combines an easy-to-use environment for surgical planning with high-end visualization and the opportunity to analyze data sets for research purposes. The demonstrator system is based on the software AMIRA. Specific algorithms for segmentation, elastic registration, and visualization have been implemented and adapted to the clinical workflow. Modules from AMIRA and the image processing library Insight Segmentation and Registration Toolkit (ITK) can be combined to solve various image processing tasks. Customized modules tailored to specific clinical problems can easily be implemented using the AMIRA application programming interface and a self-developed framework for ITK filters. Visualization is done via autostereoscopic displays, which provide a 3D impression without viewing aids. A Spaceball device allows a comfortable, intuitive way of navigation in the data sets. Via an interface to a neurosurgical navigation system, the demonstrator system can be used intraoperatively. The precision, applicability, and benefit of the demonstrator system for planning of neurosurgical interventions and for neurosurgical research were successfully evaluated by neurosurgeons using phantom and patient data sets.

  2. [3D virtual imaging of the upper airways].

    PubMed

    Ferretti, G; Coulomb, M

    2000-04-01

    The different three dimensional reconstructions of the upper airways that can be obtained with spiral computed tomograpy (CT) are presented here. The parameters indispensable to achieve as real as possible spiral CT images are recalled together with the advantages and disadvantages of the different techniues. Multislice reconstruction (MSR) produces slices in different planes of space with the high contrast of CT slices. They provide information similar to that obtained for the rare indications for thoracic MRI. Thick slice reconstructions with maximum intensity projection (MIP) or minimum intensity projection (minIP) give projection views where the contrast can be modified by selecting the more dense (MIP) or less dense (minIP) voxels. They find their application in the exploration of the upper airways. Surface and volume external 3D reconstructions can be obtained. They give an overall view of the upper airways, similar to a bronchogram. Virtual endoscopy reproduces real endoscopic images but cannot provide information on the aspect of the mucosa or biopsy specimens. It offers possible applications for preparing, guiding and controlling interventional fibroscopy procedures.

  3. Multiframe image point matching and 3-d surface reconstruction.

    PubMed

    Tsai, R Y

    1983-02-01

    This paper presents two new methods, the Joint Moment Method (JMM) and the Window Variance Method (WVM), for image matching and 3-D object surface reconstruction using multiple perspective views. The viewing positions and orientations for these perspective views are known a priori, as is usually the case for such applications as robotics and industrial vision as well as close range photogrammetry. Like the conventional two-frame correlation method, the JMM and WVM require finding the extrema of 1-D curves, which are proved to theoretically approach a delta function exponentially as the number of frames increases for the JMM and are much sharper than the two-frame correlation function for both the JMM and the WVM, even when the image point to be matched cannot be easily distinguished from some of the other points. The theoretical findings have been supported by simulations. It is also proved that JMM and WVM are not sensitive to certain radiometric effects. If the same window size is used, the computational complexity for the proposed methods is about n - 1 times that for the two-frame method where n is the number of frames. Simulation results show that the JMM and WVM require smaller windows than the two-frame correlation method with better accuracy, and therefore may even be more computationally feasible than the latter since the computational complexity increases quadratically as a function of the window size.

  4. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    NASA Astrophysics Data System (ADS)

    Ranjan Gartia, Manas; Hsiao, Austin; Sivaguru, Mayandi; Chen, Yi; Logan Liu, G.

    2011-09-01

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  5. Advanced 3D polarimetric flash ladar imaging through foliage

    NASA Astrophysics Data System (ADS)

    Murray, James T.; Moran, Steven E.; Roddier, Nicolas; Vercillo, Richard; Bridges, Robert; Austin, William

    2003-08-01

    High-resolution three-dimensional flash ladar system technologies are under development that enables remote identification of vehicles and armament hidden by heavy tree canopies. We have developed a sensor architecture and design that employs a 3D flash ladar receiver to address this mission. The receiver captures 128×128×>30 three-dimensional images for each laser pulse fired. The voxel size of the image is 3"×3"×4" at the target location. A novel signal-processing algorithm has been developed that achieves sub-voxel (sub-inch) range precision estimates of target locations within each pixel. Polarization discrimination is implemented to augment the target-to-foliage contrast. When employed, this method improves the range resolution of the system beyond the classical limit (based on pulsewidth and detection bandwidth). Experiments were performed with a 6 ns long transmitter pulsewidth that demonstrate 1-inch range resolution of a tank-like target that is occluded by foliage and a range precision of 0.3" for unoccluded targets.

  6. 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

  7. 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.

  8. 3D-3D registration of partial capitate bones using spin-images

    NASA Astrophysics Data System (ADS)

    Breighner, Ryan; Holmes, David R.; Leng, Shuai; An, Kai-Nan; McCollough, Cynthia; Zhao, Kristin

    2013-03-01

    It is often necessary to register partial objects in medical imaging. Due to limited field of view (FOV), the entirety of an object cannot always be imaged. This study presents a novel application of an existing registration algorithm to this problem. The spin-image algorithm [1] creates pose-invariant representations of global shape with respect to individual mesh vertices. These `spin-images,' are then compared for two different poses of the same object to establish correspondences and subsequently determine relative orientation of the poses. In this study, the spin-image algorithm is applied to 4DCT-derived capitate bone surfaces to assess the relative accuracy of registration with various amounts of geometry excluded. The limited longitudinal coverage under the 4DCT technique (38.4mm, [2]), results in partial views of the capitate when imaging wrist motions. This study assesses the ability of the spin-image algorithm to register partial bone surfaces by artificially restricting the capitate geometry available for registration. Under IRB approval, standard static CT and 4DCT scans were obtained on a patient. The capitate was segmented from the static CT and one phase of 4DCT in which the whole bone was available. Spin-image registration was performed between the static and 4DCT. Distal portions of the 4DCT capitate (10-70%) were then progressively removed and registration was repeated. Registration accuracy was evaluated by angular errors and the percentage of sub-resolution fitting. It was determined that 60% of the distal capitate could be omitted without appreciable effect on registration accuracy using the spin-image algorithm (angular error < 1.5 degree, sub-resolution fitting < 98.4%).

  9. 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.

  10. 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.

  11. 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...

  12. 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...

  13. 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.

  14. 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.

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

    PubMed Central

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

    2016-01-01

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

  16. 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. Comparison of bootstrap resampling methods for 3-D PET imaging.

    PubMed

    Lartizien, C; Aubin, J-B; Buvat, I

    2010-07-01

    Two groups of bootstrap methods have been proposed to estimate the statistical properties of positron emission tomography (PET) images by generating multiple statistically equivalent data sets from few data samples. The first group generates resampled data based on a parametric approach assuming that data from which resampling is performed follows a Poisson distribution while the second group consists of nonparametric approaches. These methods either require a unique original sample or a series of statistically equivalent data that can be list-mode files or sinograms. Previous reports regarding these bootstrap approaches suggest different results. This work compares the accuracy of three of these bootstrap methods for 3-D PET imaging based on simulated data. Two methods are based on a unique file, namely a list-mode based nonparametric (LMNP) method and a sinogram based parametric (SP) method. The third method is a sinogram-based nonparametric (SNP) method. Another original method (extended LMNP) was also investigated, which is an extension of the LMNP methods based on deriving a resampled list-mode file by drawings events from multiple original list-mode files. Our comparison is based on the analysis of the statistical moments estimated on the repeated and resampled data. This includes the probability density function and the moments of order 1 and 2. Results show that the two methods based on multiple original data (SNP and extended LMNP) are the only methods that correctly estimate the statistical parameters. Performances of the LMNP and SP methods are variable. Simulated data used in this study were characterized by a high noise level. Differences among the tested strategies might be reduced with clinical data sets with lower noise.

  18. 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

  19. 3-D image pre-processing algorithms for improved automated tracing of neuronal arbors.

    PubMed

    Narayanaswamy, Arunachalam; Wang, Yu; Roysam, Badrinath

    2011-09-01

    The accuracy and reliability of automated neurite tracing systems is ultimately limited by image quality as reflected in the signal-to-noise ratio, contrast, and image variability. This paper describes a novel combination of image processing methods that operate on images of neurites captured by confocal and widefield microscopy, and produce synthetic images that are better suited to automated tracing. The algorithms are based on the curvelet transform (for denoising curvilinear structures and local orientation estimation), perceptual grouping by scalar voting (for elimination of non-tubular structures and improvement of neurite continuity while preserving branch points), adaptive focus detection, and depth estimation (for handling widefield images without deconvolution). The proposed methods are fast, and capable of handling large images. Their ability to handle images of unlimited size derives from automated tiling of large images along the lateral dimension, and processing of 3-D images one optical slice at a time. Their speed derives in part from the fact that the core computations are formulated in terms of the Fast Fourier Transform (FFT), and in part from parallel computation on multi-core computers. The methods are simple to apply to new images since they require very few adjustable parameters, all of which are intuitive. Examples of pre-processing DIADEM Challenge images are used to illustrate improved automated tracing resulting from our pre-processing methods.

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

    PubMed

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

    2015-01-01

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

  1. Determining the chronological order of crossing lines using 3D imaging techniques

    NASA Astrophysics Data System (ADS)

    Heikkinen, V.; Barbeau, C.; Kassamakov, I.; Lehto, S.; Reinikainen, T.; Aaltonen, J.; Hæggström, E.

    2010-10-01

    The chronological order of creation of crossing lines scratched into a copper surface was determined using 3D profiles measured with SWLI and CM. As the methods used are based only on the deformations of the surface and since the imaging techniques can be used for different materials, the proposed methods are potentially effective also on other materials. Determining the chronological order of orthogonally crossing lines is studied in forensic science. The order of creation of such lines allows in some cases determination of the history of an object without comparing it to other objects.. Methods based on two dimensional (2D) imaging have been used for this task, but such methods are ineffective if the lines are made with a similar tool. We apply Scanning White Light Interferometry (SWLI) and Confocal Microscopy (CM) to study crossing lines on a copper surface scratched with a scratching device. Both SWLI and CM quantitatively measure the 3D surface profiles with sufficient accuracy for forensic applications. 3D image processing allows removing unimportant features, such as surface form and roughness, as well as measurement noise from the measured profiles. Separating inherent features in the crossing area, from other surface characteristics allows one to determine the sequence of creation of the lines even on a rough and wavy surface.

  2. 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.

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

    PubMed

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

    2015-08-01

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

  4. 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

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

    PubMed Central

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

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

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

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

    PubMed

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

    2016-11-25

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

  8. 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.

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

    PubMed

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

    2015-09-21

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

  10. 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.

  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 Adaptive Sparsity Based Image Compression with Applications to Optical Coherence Tomography

    PubMed Central

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

    2015-01-01

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

  13. 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

  14. 3-D Imaging Systems for Agricultural Applications-A Review.

    PubMed

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

    2016-04-29

    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.

  15. 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.

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

    PubMed

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

    2014-10-01

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

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

    PubMed Central

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

    2016-01-01

    The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assemblies within such limited volumes requires precise targeting of the FIB milling process. In this study, we present the development of a cryo-stage allowing for spinning-disk confocal light microscopy at cryogenic temperatures and describe the incorporation of the new hardware into existing workflows for cellular sample preparation by cryo-FIB. Introduction of fiducial markers and subsequent computation of three-dimensional coordinate transformations provide correlation between light microscopy and scanning electron microscopy/FIB. The correlative approach is employed to guide the FIB milling process of vitrified cellular samples and to capture specific structures, namely fluorescently labeled lipid droplets, in lamellas that are 300 nm thick. The correlation procedure is then applied to localize the fluorescently labeled structures in the transmission electron microscopy image of the lamella. This approach can be employed to navigate the acquisition of cryo-ET data within FIB-lamellas at specific locations, unambiguously identified by fluorescence microscopy. PMID:26769364

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

    PubMed Central

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

    2014-01-01

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

  20. Live cell imaging by multifocal multiphoton microscopy.

    PubMed

    Straub, M; Lodemann, P; Holroyd, P; Jahn, R; Hell, S W

    2000-10-01

    Multifocal multiphoton microscopy (MMM) permits parallel multiphoton excitation by scanning an array of high numerical aperture foci across a plane in the sample. MMM is particularly suitable for live cell investigations since it combines advantages of standard multiphoton microscopy such as optical sectioning and suppression of out-of-focus phototoxicity with high recording speeds. Here we describe several applications of MMM to live cell imaging using the neuroendocrine cell line PC12 and bovine chromaffin cells. Stainings were performed with the acidophilic dye acridine orange and the lipophilic dyes FM1-43 and Fast DiA as well as by transfection of the cells with GFP. In both bovine chromaffin and PC12 cells structural elements of nuclear chromatin and the 3-D distribution of acidic organelles inside the cells were visualized. In PC12 cells differentiated by nerve growth factor examples of neurites were monitored. Stainings of membranes were used to reconstruct the morphology of cells and neurites in three dimensions by volume-rendering and by isosurface plots. 3-D reconstructions were composed from stacks of about 50 images each with a diameter of 30-100 microm that were acquired within a few seconds. We conclude that MMM proves to be a technically simple and very effective method for fast 3-D live cell imaging at high resolution.

  1. 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.

  2. High resolution 3D confocal microscope imaging of volcanic ash particles.

    PubMed

    Wertheim, David; Gillmore, Gavin; Gill, Ian; Petford, Nick

    2017-07-15

    We present initial results from a novel high resolution confocal microscopy study of the 3D surface structure of volcanic ash particles from two recent explosive basaltic eruptions, Eyjafjallajökull (2010) and Grimsvötn (2011), in Iceland. The majority of particles imaged are less than 100μm in size and include PM10s, known to be harmful to humans if inhaled. Previous studies have mainly used 2D microscopy to examine volcanic particles. The aim of this study was to test the potential of 3D laser scanning confocal microscopy as a reliable analysis tool for these materials and if so to what degree high resolution surface and volume data could be obtained that would further aid in their classification. First results obtained using an Olympus LEXT scanning confocal microscope with a ×50 and ×100 objective lens are highly encouraging. They reveal a range of discrete particle types characterised by sharp or concave edges consistent with explosive formation and sudden rupture of magma. Initial surface area/volume ratios are given that may prove useful in subsequent modelling of damage to aircraft engines and human tissue where inhalation has occurred.

  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. High-Performance 3D Image Processing Architectures for Image-Guided Interventions

    DTIC Science & Technology

    2008-01-01

    Circuits and Systems, vol. 1 (2), 2007, pp. 116-127. iv • O. Dandekar, C. Castro- Pareja , and R. Shekhar, “FPGA-based real-time 3D image...How low can we go?,” presented at IEEE International Symposium on Biomedical Imaging, 2006, pp. 502-505. • C. R. Castro- Pareja , O. Dandekar, and R...Venugopal, C. R. Castro- Pareja , and O. Dandekar, “An FPGA-based 3D image processor with median and convolution filters for real-time applications,” in

  5. Robust Reconstruction and Generalized Dual Hahn Moments Invariants Extraction for 3D Images

    NASA Astrophysics Data System (ADS)

    Mesbah, Abderrahim; Zouhri, Amal; El Mallahi, Mostafa; Zenkouar, Khalid; Qjidaa, Hassan

    2017-03-01

    In this paper, we introduce a new set of 3D weighed dual Hahn moments which are orthogonal on a non-uniform lattice and their polynomials are numerically stable to scale, consequent, producing a set of weighted orthonormal polynomials. The dual Hahn is the general case of Tchebichef and Krawtchouk, and the orthogonality of dual Hahn moments eliminates the numerical approximations. The computational aspects and symmetry property of 3D weighed dual Hahn moments are discussed in details. To solve their inability to invariability of large 3D images, which cause to overflow issues, a generalized version of these moments noted 3D generalized weighed dual Hahn moment invariants are presented where whose as linear combination of regular geometric moments. For 3D pattern recognition, a generalized expression of 3D weighted dual Hahn moment invariants, under translation, scaling and rotation transformations, have been proposed where a new set of 3D-GWDHMIs have been provided. In experimental studies, the local and global capability of free and noisy 3D image reconstruction of the 3D-WDHMs has been compared with other orthogonal moments such as 3D Tchebichef and 3D Krawtchouk moments using Princeton Shape Benchmark database. On pattern recognition using the 3D-GWDHMIs like 3D object descriptors, the experimental results confirm that the proposed algorithm is more robust than other orthogonal moments for pattern classification of 3D images with and without noise.

  6. 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.

  7. Potential Cost Savings for Use of 3D Printing Combined With 3D Imaging and CPLM for Fleet Maintenance and Revitalization

    DTIC Science & Technology

    2013-12-04

    pmlkploba=obmloq=pbofbp= = = Potential Cost Savings for Use of 3D Printing Combined With 3D Imaging and CPLM for Fleet Maintenance and Revitalization...REPORT TYPE 3. DATES COVERED 00-00-2013 to 00-00-2013 4. TITLE AND SUBTITLE Potential Cost Savings for Use of 3D Printing Combined With 3D ...oÉëÉ~êÅÜ=mêçÖê~ã= ëéçåëçêÉÇ=oÉéçêí=pÉêáÉë= Potential Cost Savings for Use of 3D Printing Combined With 3D Imaging and CPLM for Fleet Maintenance and

  8. 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.

  9. 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.

  10. Evolution of 3D surface imaging systems in facial plastic surgery.

    PubMed

    Tzou, Chieh-Han John; Frey, Manfred

    2011-11-01

    Recent advancements in computer technologies have propelled the development of 3D imaging systems. 3D surface-imaging is taking surgeons to a new level of communication with patients; moreover, it provides quick and standardized image documentation. This article recounts the chronologic evolution of 3D surface imaging, and summarizes the current status of today's facial surface capturing technology. This article also discusses current 3D surface imaging hardware and software, and their different techniques, technologies, and scientific validation, which provides surgeons with the background information necessary for evaluating the systems and knowledge about the systems they might incorporate into their own practice.

  11. 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.

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

    PubMed Central

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

    2014-01-01

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

  13. 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

  14. 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.

  15. 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.

  16. Chip-scale microscopy imaging.

    PubMed

    Zheng, Guoan

    2012-08-01

    Chip-scale microscopy imaging platforms are pivotal for improving the efficiency of modern biomedical and bioscience experiments. Their integration with other lab-on-a-chip techniques would allow rapid, reliable and high-throughput sample analysis for applications in diverse disciplines. In typical chip-scale microscopy imaging platforms, the light path can be generalized to the following steps: photons leave the light source, interact with the sample and finally are detected by the sensor. Based on the light path of these platforms, the current review aims to provide some insights on design strategies for chip-scale microscopy. Specifically, we analyze current chip-scale microscopy approaches from three aspects: illumination design, sample manipulation and substrate/imager modification. We also discuss some opportunities for future developments of chip-scale microscopy, such as time multiplexed structured illumination and hydrodynamic focusing for high throughput sample manipulation.

  17. Volumetric medical image compression using 3D listless embedded block partitioning.

    PubMed

    Senapati, Ranjan K; Prasad, P M K; Swain, Gandharba; Shankar, T N

    2016-01-01

    This paper presents a listless variant of a modified three-dimensional (3D)-block coding algorithm suitable for medical image compression. A higher degree of correlation is achieved by using a 3D hybrid transform. The 3D hybrid transform is performed by a wavelet transform in the spatial dimension and a Karhunen-Loueve transform in the spectral dimension. The 3D transformed coefficients are arranged in a one-dimensional (1D) fashion, as in the hierarchical nature of the wavelet-coefficient distribution strategy. A novel listless block coding algorithm is applied to the mapped 1D coefficients which encode in an ordered-bit-plane fashion. The algorithm originates from the most significant bit plane and terminates at the least significant bit plane to generate an embedded bit stream, as in 3D-SPIHT. The proposed algorithm is called 3D hierarchical listless block (3D-HLCK), which exhibits better compression performance than that exhibited by 3D-SPIHT. Further, it is highly competitive with some of the state-of-the-art 3D wavelet coders for a wide range of bit rates for magnetic resonance, digital imaging and communication in medicine and angiogram images. 3D-HLCK provides rate and resolution scalability similar to those provided by 3D-SPIHT and 3D-SPECK. In addition, a significant memory reduction is achieved owing to the listless nature of 3D-HLCK.

  18. Image guidance of breast cancer surgery using 3-D ultrasound images and augmented reality visualization.

    PubMed

    Sato, Y; Nakamoto, M; Tamaki, Y; Sasama, T; Sakita, I; Nakajima, Y; Monden, M; Tamura, S

    1998-10-01

    This paper describes augmented reality visualization for the guidance of breast-conservative cancer surgery using ultrasonic images acquired in the operating room just before surgical resection. By combining an optical three-dimensional (3-D) position sensor, the position and orientation of each ultrasonic cross section are precisely measured to reconstruct geometrically accurate 3-D tumor models from the acquired ultrasonic images. Similarly, the 3-D position and orientation of a video camera are obtained to integrate video and ultrasonic images in a geometrically accurate manner. Superimposing the 3-D tumor models onto live video images of the patient's breast enables the surgeon to perceive the exact 3-D position of the tumor, including irregular cancer invasions which cannot be perceived by touch, as if it were visible through the breast skin. Using the resultant visualization, the surgeon can determine the region for surgical resection in a more objective and accurate manner, thereby minimizing the risk of a relapse and maximizing breast conservation. The system was shown to be effective in experiments using phantom and clinical data.

  19. 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

    2016-12-01

    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.

  20. 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.

  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. 3D imaging of telomeres and nuclear architecture: An emerging tool of 3D nano-morphology-based diagnosis.

    PubMed

    Knecht, Hans; Mai, Sabine

    2011-04-01

    Patient samples are evaluated by experienced pathologists whose diagnosis guides treating physicians. Pathological diagnoses are complex and often assisted by the application of specific tissue markers. However, cases still exist where pathologists cannot distinguish between closely related entities or determine the aggressiveness of the disease they identify under the microscope. This is due to the absence of reliable markers that define diagnostic subgroups in several cancers. Three-dimensional (3D) imaging of nuclear telomere signatures is emerging as a new tool that may change this situation offering new opportunities to the patients. This article will review current and future avenues in the assessment of diagnostic patient samples.

  3. 3D nanostructure reconstruction based on the SEM imaging principle, and applications.

    PubMed

    Zhu, Fu-Yun; Wang, Qi-Qi; Zhang, Xiao-Sheng; Hu, Wei; Zhao, Xin; Zhang, Hai-Xia

    2014-05-09

    This paper addresses a novel 3D reconstruction method for nanostructures based on the scanning electron microscopy (SEM) imaging principle. In this method, the shape from shading (SFS) technique is employed, to analyze the gray-scale information of a single top-view SEM image which contains all the visible surface information, and finally to reconstruct the 3D surface morphology. It offers not only unobstructed observation from various angles but also the exact physical dimensions of nanostructures. A convenient and commercially available tool (NanoViewer) is developed based on this method for nanostructure analysis and characterization of properties. The reconstruction result coincides well with the SEM nanostructure image and is verified in different ways. With the extracted structure information, subsequent research of the nanostructure can be carried out, such as roughness analysis, optimizing properties by structure improvement and performance simulation with a reconstruction model. Efficient, practical and non-destructive, the method will become a powerful tool for nanostructure surface observation and characterization.

  4. 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.

  5. 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

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

    PubMed Central

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

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2010-02-01

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

  8. 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.

  9. Holographic imaging of 3D objects on dichromated polymer systems

    NASA Astrophysics Data System (ADS)

    Lemelin, Guylain; Jourdain, Anne; Manivannan, Gurusamy; Lessard, Roger A.

    1996-01-01

    Conventional volume transmission holograms of a 3D scene were recorded on dichromated poly(acrylic acid) (DCPAA) films under 488 nm light. The holographic characterization and quality of reconstruction have been studied by varying the influencing parameters such as concentration of dichromate and electron donor, and the molecular weight of the polymer matrix. Ammonium and potassium dichromate have been employed to sensitize the poly(acrylic) matrix. the recorded hologram can be efficiently reconstructed either with red light or with low energy in the blue region without any post thermal or chemical processing.

  10. 3-D Velocity Measurement of Natural Convection Using Image Processing

    NASA Astrophysics Data System (ADS)

    Shinoki, Masatoshi; Ozawa, Mamoru; Okada, Toshifumi; Kimura, Ichiro

    This paper describes quantitative three-dimensional measurement method for flow field of a rotating Rayleigh-Benard convection in a cylindrical cell heated below and cooled above. A correlation method for two-dimensional measurement was well advanced to a spatio-temporal correlation method. Erroneous vectors, often appeared in the correlation method, was successfully removed using Hopfield neural network. As a result, calculated 3-D velocity vector distribution well corresponded to the observed temperature distribution. Consequently, the simultaneous three-dimensional measurement system for temperature and flow field was developed.

  11. 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

  12. 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.

  13. 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.

  14. Multi-layer 3D imaging using a few viewpoint images and depth map

    NASA Astrophysics Data System (ADS)

    Suginohara, Hidetsugu; Sakamoto, Hirotaka; Yamanaka, Satoshi; Suyama, Shiro; Yamamoto, Hirotsugu

    2015-03-01

    In this paper, we propose a new method that makes multi-layer images from a few viewpoint images to display a 3D image by the autostereoscopic display that has multiple display screens in the depth direction. We iterate simple "Shift and Subtraction" processes to make each layer image alternately. The image made in accordance with depth map like a volume slicing by gradations is used as the initial solution of iteration process. Through the experiments using the prototype stacked two LCDs, we confirmed that it was enough to make multi-layer images from three viewpoint images to display a 3D image. Limiting the number of viewpoint images, the viewing area that allows stereoscopic view becomes narrow. To broaden the viewing area, we track the head motion of the viewer and update screen images in real time so that the viewer can maintain correct stereoscopic view within +/- 20 degrees area. In addition, we render pseudo multiple viewpoint images using depth map, then we can generate motion parallax at the same time.

  15. Analysis of fracture networks in a reservoir dolomite by 3D micro-imaging

    NASA Astrophysics Data System (ADS)

    Voorn, Maarten; Hoyer, Stefan; Exner, Ulrike; Reuschlé, Thierry

    2013-04-01

    the fracture apertures can be defined. One can use an adaptation to the filtering technique to determine the orientation of the extracted fractures in a sample as well. All these analyses contribute to comprehensive information on the fracture network in a sample. This processed data can then serve as input for permeability modelling. All results from the µCT imaging and data analyses are combined and cross-calibrated with information from different techniques, obtained on the same samples. We apply for example the aforementioned laboratory methods (including permeability determination under increasing confining pressure) and 2D thin section analysis. Furthermore, we use 3D Focussed Ion Beam - Scanning Electron Microscopy (FIB-SEM) tomography to obtain 3D information at a much smaller scale. Eventually, we try to obtain an as complete as possible set of information for every sample analysed. The combination of the various techniques shows that especially the 3D imaging is important, as even on the small sample sizes, large differences between the present fracture networks can be observed. This starts to make it possible to explain the reasons for found differences between various wells. This research thereby hopefully leads to a better understanding of the (microscale) characteristics of a reservoir system.

  16. 3D reconstruction and characterization of laser induced craters by in situ optical microscopy

    NASA Astrophysics Data System (ADS)

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

    2016-06-01

    A low-cost optical microscope was developed and coupled to an irradiation system in order to study the induced effects on material during a multipulse regime by an in situ visual inspection of the surface, in particular of the spot generated at different pulses. In the case of laser ablation, a reconstruction of the crater in 3D was made from the images of the sample surface taken during the irradiation process, and the subsequent profiles of ablated material were extracted. The implementation of this homemade optical device gives an added value to the irradiation system, providing information about morphology evolution of irradiated area when successive pulses are applied. In particular, the determination of ablatio