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Sample records for super resolution microscopy

  1. The 2015 super-resolution microscopy roadmap

    International Nuclear Information System (INIS)

    Hell, Stefan W; Sahl, Steffen J; Bates, Mark; Jakobs, Stefan; Zhuang, Xiaowei; Heintzmann, Rainer; Booth, Martin J; Bewersdorf, Joerg; Shtengel, Gleb; Hess, Harald; Tinnefeld, Philip; Honigmann, Alf; Testa, Ilaria; Cognet, Laurent; Lounis, Brahim; Ewers, Helge; Davis, Simon J; Eggeling, Christian; Klenerman, David; Willig, Katrin I

    2015-01-01

    Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio)physical and biomedical research, particularly with respect to the study of living cells and organisms. Unfortunately, the applicability of the optical microscope is limited, since the diffraction of light imposes limitations on the spatial resolution of the image. Consequently the details of, for example, cellular protein distributions, can be visualized only to a certain extent. Fortunately, recent years have witnessed the development of ‘super-resolution’ far-field optical microscopy (nanoscopy) techniques such as stimulated emission depletion (STED), ground state depletion (GSD), reversible saturated optical (fluorescence) transitions (RESOLFT), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) or saturated structured illumination microscopy (SSIM), all in one way or another addressing the problem of the limited spatial resolution of far-field optical microscopy. While SIM achieves a two-fold improvement in spatial resolution compared to conventional optical microscopy, STED, RESOLFT, PALM/STORM, or SSIM have all gone beyond, pushing the limits of optical image resolution to the nanometer scale. Consequently, all super-resolution techniques open new avenues of biomedical research. Because the field is so young, the potential capabilities of different super-resolution microscopy approaches have yet to be fully explored, and uncertainties remain when considering the best choice of methodology. Thus, even for experts, the road to the future is sometimes shrouded in mist. The super-resolution optical microscopy roadmap of Journal of Physics D: Applied Physics addresses this need for clarity. It provides guidance to the outstanding questions through a collection of short review articles from experts in the field, giving a thorough

  2. Shedding light on endocytosis with optimized super-resolution microscopy

    NARCIS (Netherlands)

    Leyton Puig, D.M.

    2017-01-01

    Super-resolution microscopy is a relatively new microscopy technique that is still under optimization. In this thesis we focus on the improvement of the quality of super-resolution images, to apply them to the study of the processes of cell signaling and endocytosis. First, we show that the use of a

  3. Example-Based Super-Resolution Fluorescence Microscopy.

    Science.gov (United States)

    Jia, Shu; Han, Boran; Kutz, J Nathan

    2018-04-23

    Capturing biological dynamics with high spatiotemporal resolution demands the advancement in imaging technologies. Super-resolution fluorescence microscopy offers spatial resolution surpassing the diffraction limit to resolve near-molecular-level details. While various strategies have been reported to improve the temporal resolution of super-resolution imaging, all super-resolution techniques are still fundamentally limited by the trade-off associated with the longer image acquisition time that is needed to achieve higher spatial information. Here, we demonstrated an example-based, computational method that aims to obtain super-resolution images using conventional imaging without increasing the imaging time. With a low-resolution image input, the method provides an estimate of its super-resolution image based on an example database that contains super- and low-resolution image pairs of biological structures of interest. The computational imaging of cellular microtubules agrees approximately with the experimental super-resolution STORM results. This new approach may offer potential improvements in temporal resolution for experimental super-resolution fluorescence microscopy and provide a new path for large-data aided biomedical imaging.

  4. Super-resolution fluorescence microscopy by stepwise optical saturation

    Science.gov (United States)

    Zhang, Yide; Nallathamby, Prakash D.; Vigil, Genevieve D.; Khan, Aamir A.; Mason, Devon E.; Boerckel, Joel D.; Roeder, Ryan K.; Howard, Scott S.

    2018-01-01

    Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the super-resolution microscopy is not feasible in many applications. In this paper, we propose and demonstrate a saturation-based super-resolution fluorescence microscopy technique that can be easily implemented and requires neither additional hardware nor complex post-processing. The method is based on the principle of stepwise optical saturation (SOS), where M steps of raw fluorescence images are linearly combined to generate an image with a M-fold increase in resolution compared with conventional diffraction-limited images. For example, linearly combining (scaling and subtracting) two images obtained at regular powers extends the resolution by a factor of 1.4 beyond the diffraction limit. The resolution improvement in SOS microscopy is theoretically infinite but practically is limited by the signal-to-noise ratio. We perform simulations and experimentally demonstrate super-resolution microscopy with both one-photon (confocal) and multiphoton excitation fluorescence. We show that with the multiphoton modality, the SOS microscopy can provide super-resolution imaging deep in scattering samples. PMID:29675306

  5. Super-resolution Microscopy in Plant Cell Imaging.

    Science.gov (United States)

    Komis, George; Šamajová, Olga; Ovečka, Miroslav; Šamaj, Jozef

    2015-12-01

    Although the development of super-resolution microscopy methods dates back to 1994, relevant applications in plant cell imaging only started to emerge in 2010. Since then, the principal super-resolution methods, including structured-illumination microscopy (SIM), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), and stimulated emission depletion microscopy (STED), have been implemented in plant cell research. However, progress has been limited due to the challenging properties of plant material. Here we summarize the basic principles of existing super-resolution methods and provide examples of applications in plant science. The limitations imposed by the nature of plant material are reviewed and the potential for future applications in plant cell imaging is highlighted. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Application of super-resolution optical microscopy in biology

    International Nuclear Information System (INIS)

    Mao Xiuhai; Du Jiancong; Huang Qing; Fan Chunhai; Deng Suhui

    2013-01-01

    Background: A noninvasive, real-time far-field optical microscopy is needed to study the dynamic function inside cells and proteins. However, the resolution limit of traditional optical microscope is about 200 nm due to the diffraction limit of light. So, it's hard to directly observe the subcellular structures. Over the past several years of microscopy development, the diffraction limit of fluorescence microscopy has been overcome and its resolution limit is about tens of nanometers. Methods: To overcome the diffraction limit of light, many super-resolution fluoresce microscopes, including stimulated emission of depletion microscopy (STED), photoactivation localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), have been developed. Conclusions: These methods have been applied in cell biology, microbiology and neurobiology, and the technology of super-resolution provides a new insight into the life science. (authors)

  7. Aberrations and adaptive optics in super-resolution microscopy

    Science.gov (United States)

    Booth, Martin; Andrade, Débora; Burke, Daniel; Patton, Brian; Zurauskas, Mantas

    2015-01-01

    As one of the most powerful tools in the biological investigation of cellular structures and dynamic processes, fluorescence microscopy has undergone extraordinary developments in the past decades. The advent of super-resolution techniques has enabled fluorescence microscopy – or rather nanoscopy – to achieve nanoscale resolution in living specimens and unravelled the interior of cells with unprecedented detail. The methods employed in this expanding field of microscopy, however, are especially prone to the detrimental effects of optical aberrations. In this review, we discuss how super-resolution microscopy techniques based upon single-molecule switching, stimulated emission depletion and structured illumination each suffer from aberrations in different ways that are dependent upon intrinsic technical aspects. We discuss the use of adaptive optics as an effective means to overcome this problem. PMID:26124194

  8. Correlative Super-Resolution Microscopy: New Dimensions and New Opportunities.

    Science.gov (United States)

    Hauser, Meghan; Wojcik, Michal; Kim, Doory; Mahmoudi, Morteza; Li, Wan; Xu, Ke

    2017-06-14

    Correlative microscopy, the integration of two or more microscopy techniques performed on the same sample, produces results that emphasize the strengths of each technique while offsetting their individual weaknesses. Light microscopy has historically been a central method in correlative microscopy due to its widespread availability, compatibility with hydrated and live biological samples, and excellent molecular specificity through fluorescence labeling. However, conventional light microscopy can only achieve a resolution of ∼300 nm, undercutting its advantages in correlations with higher-resolution methods. The rise of super-resolution microscopy (SRM) over the past decade has drastically improved the resolution of light microscopy to ∼10 nm, thus creating exciting new opportunities and challenges for correlative microscopy. Here we review how these challenges are addressed to effectively correlate SRM with other microscopy techniques, including light microscopy, electron microscopy, cryomicroscopy, atomic force microscopy, and various forms of spectroscopy. Though we emphasize biological studies, we also discuss the application of correlative SRM to materials characterization and single-molecule reactions. Finally, we point out current limitations and discuss possible future improvements and advances. We thus demonstrate how a correlative approach adds new dimensions of information and provides new opportunities in the fast-growing field of SRM.

  9. Graphene-enabled electron microscopy and correlated super-resolution microscopy of wet cells.

    Science.gov (United States)

    Wojcik, Michal; Hauser, Margaret; Li, Wan; Moon, Seonah; Xu, Ke

    2015-06-11

    The application of electron microscopy to hydrated biological samples has been limited by high-vacuum operating conditions. Traditional methods utilize harsh and laborious sample dehydration procedures, often leading to structural artefacts and creating difficulties for correlating results with high-resolution fluorescence microscopy. Here, we utilize graphene, a single-atom-thick carbon meshwork, as the thinnest possible impermeable and conductive membrane to protect animal cells from vacuum, thus enabling high-resolution electron microscopy of wet and untreated whole cells with exceptional ease. Our approach further allows for facile correlative super-resolution and electron microscopy of wet cells directly on the culturing substrate. In particular, individual cytoskeletal actin filaments are resolved in hydrated samples through electron microscopy and well correlated with super-resolution results.

  10. Application of spectroscopy and super-resolution microscopy: Excited state

    Energy Technology Data Exchange (ETDEWEB)

    Bhattacharjee, Ujjal [Iowa State Univ., Ames, IA (United States)

    2016-02-19

    Photophysics of inorganic materials and organic molecules in complex systems have been extensively studied with absorption and emission spectroscopy.1-4 Steady-state and time-resolved fluorescence studies are commonly carried out to characterize excited-state properties of fluorophores. Although steady-state fluorescence measurements are widely used for analytical applications, time-resolved fluorescence measurements provide more detailed information about excited-state properties and the environment in the vicinity of the fluorophore. Many photophysical processes, such as photoinduced electron transfer (PET), rotational reorientation, solvent relaxation, and energy transfer, occur on a nanosecond (10-9 s) timescale, thus affecting the lifetime of the fluorophores. Moreover, time-resolved microscopy methods, such as lifetimeimaging, combine the benefits of the microscopic measurement and information-rich, timeresolved data. Thus, time-resolved fluorescence spectroscopy combined with microscopy can be used to quantify these processes and to obtain a deeper understanding of the chemical surroundings of the fluorophore in a small area under investigation. This thesis discusses various photophysical and super-resolution microscopic studies of organic and inorganic materials, which have been outlined below.

  11. Solid-immersion fluorescence microscopy with increased emission and super resolution

    Energy Technology Data Exchange (ETDEWEB)

    Liau, Z. L.; Porter, J. M. [Lincoln Laboratory, Massachusetts Institute of Technology, Lexington, Massachusetts 02420 (United States); Liau, A. A.; Chen, J. J. [Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States); Salmon, W. C. [Whitehead Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States); Sheu, S. S. [Department of Medicine, Jefferson Medical College, Philadelphia, Pennsylvania 19107 (United States)

    2015-01-07

    We investigate solid-immersion fluorescence microscopy suitable for super-resolution nanotechnology and biological imaging, and have observed limit of resolution as small as 15 nm with microspheres, mitochondria, and chromatin fibers. We have further observed that fluorescence efficiency increases with excitation power density, implicating appreciable stimulated emission and increased resolution. We discuss potential advantages of the solid-immersion microscopy, including combined use with previously established super-resolution techniques for reaching deeper beyond the conventional diffraction limit.

  12. Microsphere-aided optical microscopy and its applications for super-resolution imaging

    Science.gov (United States)

    Upputuri, Paul Kumar; Pramanik, Manojit

    2017-12-01

    The spatial resolution of a standard optical microscope (SOM) is limited by diffraction. In visible spectrum, SOM can provide ∼ 200 nm resolution. To break the diffraction limit several approaches were developed including scanning near field microscopy, metamaterial super-lenses, nanoscale solid immersion lenses, super-oscillatory lenses, confocal fluorescence microscopy, techniques that exploit non-linear response of fluorophores like stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, etc. Recently, photonic nanojet generated by a dielectric microsphere was used to break the diffraction limit. The microsphere-approach is simple, cost-effective and can be implemented under a standard microscope, hence it has gained enormous attention for super-resolution imaging. In this article, we briefly review the microsphere approach and its applications for super-resolution imaging in various optical imaging modalities.

  13. Super-resolution microscopy in studying neuroendocrine cell function

    Directory of Open Access Journals (Sweden)

    Anneka eBost

    2013-11-01

    Full Text Available The last two decades have seen a tremendous development in high resolution microscopy techniques giving rise to acronyms such as TIRFM, SIM, PALM, STORM, and STED. The goal of all these techniques is to overcome the physical resolution barrier of light microscopy in order to resolve precise protein localization and possibly their interaction in cells. Neuroendocrine cell function is to secrete hormones and peptides on demand. This fine-tuned multi-step process is mediated by a large array of proteins. Here, we review the new microscopy techniques used to obtain high resolution and how they have been applied to increase our knowledge of the molecular mechanisms involved in neuroendocrine cell secretion. Further the limitations of these methods are discussed and insights in possible new applications are provided.

  14. A Microfluidic Platform for Correlative Live-Cell and Super-Resolution Microscopy

    Science.gov (United States)

    Tam, Johnny; Cordier, Guillaume Alan; Bálint, Štefan; Sandoval Álvarez, Ángel; Borbely, Joseph Steven; Lakadamyali, Melike

    2014-01-01

    Recently, super-resolution microscopy methods such as stochastic optical reconstruction microscopy (STORM) have enabled visualization of subcellular structures below the optical resolution limit. Due to the poor temporal resolution, however, these methods have mostly been used to image fixed cells or dynamic processes that evolve on slow time-scales. In particular, fast dynamic processes and their relationship to the underlying ultrastructure or nanoscale protein organization cannot be discerned. To overcome this limitation, we have recently developed a correlative and sequential imaging method that combines live-cell and super-resolution microscopy. This approach adds dynamic background to ultrastructural images providing a new dimension to the interpretation of super-resolution data. However, currently, it suffers from the need to carry out tedious steps of sample preparation manually. To alleviate this problem, we implemented a simple and versatile microfluidic platform that streamlines the sample preparation steps in between live-cell and super-resolution imaging. The platform is based on a microfluidic chip with parallel, miniaturized imaging chambers and an automated fluid-injection device, which delivers a precise amount of a specified reagent to the selected imaging chamber at a specific time within the experiment. We demonstrate that this system can be used for live-cell imaging, automated fixation, and immunostaining of adherent mammalian cells in situ followed by STORM imaging. We further demonstrate an application by correlating mitochondrial dynamics, morphology, and nanoscale mitochondrial protein distribution in live and super-resolution images. PMID:25545548

  15. Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane.

    Science.gov (United States)

    Stone, Matthew B; Shelby, Sarah A; Veatch, Sarah L

    2017-06-14

    Lipids and the membranes they form are fundamental building blocks of cellular life, and their geometry and chemical properties distinguish membranes from other cellular environments. Collective processes occurring within membranes strongly impact cellular behavior and biochemistry, and understanding these processes presents unique challenges due to the often complex and myriad interactions between membrane components. Super-resolution microscopy offers a significant gain in resolution over traditional optical microscopy, enabling the localization of individual molecules even in densely labeled samples and in cellular and tissue environments. These microscopy techniques have been used to examine the organization and dynamics of plasma membrane components, providing insight into the fundamental interactions that determine membrane functions. Here, we broadly introduce the structure and organization of the mammalian plasma membrane and review recent applications of super-resolution microscopy to the study of membranes. We then highlight some inherent challenges faced when using super-resolution microscopy to study membranes, and we discuss recent technical advancements that promise further improvements to super-resolution microscopy and its application to the plasma membrane.

  16. Super-resolution optical microscopy for studying membrane structure and dynamics.

    Science.gov (United States)

    Sezgin, Erdinc

    2017-07-12

    Investigation of cell membrane structure and dynamics requires high spatial and temporal resolution. The spatial resolution of conventional light microscopy is limited due to the diffraction of light. However, recent developments in microscopy enabled us to access the nano-scale regime spatially, thus to elucidate the nanoscopic structures in the cellular membranes. In this review, we will explain the resolution limit, address the working principles of the most commonly used super-resolution microscopy techniques and summarise their recent applications in the biomembrane field.

  17. An introduction to optical super-resolution microscopy for the adventurous biologist

    Science.gov (United States)

    Vangindertael, J.; Camacho, R.; Sempels, W.; Mizuno, H.; Dedecker, P.; Janssen, K. P. F.

    2018-04-01

    Ever since the inception of light microscopy, the laws of physics have seemingly thwarted every attempt to visualize the processes of life at its most fundamental, sub-cellular, level. The diffraction limit has restricted our view to length scales well above 250 nm and in doing so, severely compromised our ability to gain true insights into many biological systems. Fortunately, continuous advancements in optics, electronics and mathematics have since provided the means to once again make physics work to our advantage. Even though some of the fundamental concepts enabling super-resolution light microscopy have been known for quite some time, practically feasible implementations have long remained elusive. It should therefore not come as a surprise that the 2014 Nobel Prize in Chemistry was awarded to the scientists who, each in their own way, contributed to transforming super-resolution microscopy from a technological tour de force to a staple of the biologist’s toolkit. By overcoming the diffraction barrier, light microscopy could once again be established as an indispensable tool in an age where the importance of understanding life at the molecular level cannot be overstated. This review strives to provide the aspiring life science researcher with an introduction to optical microscopy, starting from the fundamental concepts governing compound and fluorescent confocal microscopy to the current state-of-the-art of super-resolution microscopy techniques and their applications.

  18. Three-dimensional super-resolution imaging for fluorescence emission difference microscopy

    Energy Technology Data Exchange (ETDEWEB)

    You, Shangting; Kuang, Cuifang, E-mail: cfkuang@zju.edu.cn; Li, Shuai; Liu, Xu; Ding, Zhihua [State key laboratory of modern optical instrumentations, Zhejiang University, Hangzhou 310027 (China)

    2015-08-15

    We propose a method theoretically to break the diffraction limit and to improve the resolution in all three dimensions for fluorescence emission difference microscopy. We produce two kinds of hollow focal spot by phase modulation. By incoherent superposition, these two kinds of focal spot yield a 3D hollow focal spot. The optimal proportion of these two kinds of spot is given in the paper. By employing 3D hollow focal spot, super-resolution image can be yielded by means of fluorescence emission difference microscopy, with resolution enhanced both laterally and axially. According to computation result, size of point spread function of three-dimensional super-resolution imaging is reduced by about 40% in all three spatial directions with respect to confocal imaging.

  19. Super-resolution microscopy reveals cell wall dynamics and peptidoglycan architecture in ovococcal bacteria.

    Science.gov (United States)

    Wheeler, Richard; Mesnage, Stéphane; Boneca, Ivo G; Hobbs, Jamie K; Foster, Simon J

    2011-12-01

    Cell morphology and viability in Eubacteria is dictated by the architecture of peptidoglycan, the major and essential structural component of the cell wall. Although the biochemical composition of peptidoglycan is well understood, how the peptidoglycan architecture can accommodate the dynamics of growth and division while maintaining cell shape remains largely unknown. Here, we elucidate the peptidoglycan architecture and dynamics of bacteria with ovoid cell shape (ovococci), which includes a number of important pathogens, by combining biochemical analyses with atomic force and super-resolution microscopies. Atomic force microscopy analysis showed preferential orientation of the peptidoglycan network parallel to the short axis of the cell, with distinct architectural features associated with septal and peripheral wall synthesis. Super-resolution three-dimensional structured illumination fluorescence microscopy was applied for the first time in bacteria to unravel the dynamics of peptidoglycan assembly in ovococci. The ovococci have a unique peptidoglycan architecture and growth mode not observed in other model organisms. © 2011 Blackwell Publishing Ltd.

  20. Towards single molecule biosensors using super-resolution fluorescence microscopy.

    Science.gov (United States)

    Lu, Xun; Nicovich, Philip R; Gaus, Katharina; Gooding, J Justin

    2017-07-15

    Conventional immunosensors require many binding events to give a single transducer output which represents the concentration of the analyte in the sample. Because of the requirements to selectively detect species in complex samples, immunosensing interfaces must allow immobilisation of antibodies while repelling nonspecific adsorption of other species. These requirements lead to quite sophisticated interfacial design, often with molecular level control, but we have no tools to characterise how well these interfaces work at the molecular level. The work reported herein is an initial feasibility study to show that antibody-antigen binding events can be monitored at the single molecule level using single molecule localisation microscopy (SMLM). The steps to achieve this first requires showing that indium tin oxide surfaces can be used for SMLM, then that these surfaces can be modified with self-assembled monolayers using organophosphonic acid derivatives, that the amount of antigens and antibodies on the surface can be controlled and monitored at the single molecule level and finally antibody binding to antigen modified surfaces can be monitored. The results show the amount of antibody that binds to an antigen modified surface is dependent on both the concentration of antigen on the surface and the concentration of antibody in solution. This study demonstrates the potential of SMLM for characterising biosensing interfaces and as the transducer in a massively parallel, wide field, single molecule detection scheme for quantitative analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. The tetraspanin web revisited by super-resolution microscopy.

    Science.gov (United States)

    Zuidscherwoude, Malou; Göttfert, Fabian; Dunlock, Vera Marie E; Figdor, Carl G; van den Bogaart, Geert; van Spriel, Annemiek B

    2015-07-17

    The spatial organization of membrane proteins in the plasma membrane is critical for signal transduction, cell communication and membrane trafficking. Tetraspanins organize functional higher-order protein complexes called 'tetraspanin-enriched microdomains (TEMs)' via interactions with partner molecules and other tetraspanins. Still, the nanoscale organization of TEMs in native plasma membranes has not been resolved. Here, we elucidated the size, density and distribution of TEMs in the plasma membrane of human B cells and dendritic cells using dual color stimulated emission depletion (STED) microscopy. We demonstrate that tetraspanins form individual nanoclusters smaller than 120 nm and quantified that a single tetraspanin CD53 cluster contains less than ten CD53 molecules. CD53 and CD37 domains were adjacent to and displayed only minor overlap with clusters containing tetraspanins CD81 or CD82. Moreover, CD53 and CD81 were found in closer proximity to their partners MHC class II and CD19 than to other tetraspanins. Although these results indicate that tetraspanin domains are adjacently positioned in the plasma membrane, they challenge the current view of the tetraspanin web of multiple tetraspanin species organized into a single domain. This study increases the molecular understanding of TEMs at the nanoscale level which is essential for comprehending tetraspanin function in cell biology.

  2. Imaging cellular structures in super-resolution with SIM, STED and Localisation Microscopy: A practical comparison.

    Science.gov (United States)

    Wegel, Eva; Göhler, Antonia; Lagerholm, B Christoffer; Wainman, Alan; Uphoff, Stephan; Kaufmann, Rainer; Dobbie, Ian M

    2016-06-06

    Many biological questions require fluorescence microscopy with a resolution beyond the diffraction limit of light. Super-resolution methods such as Structured Illumination Microscopy (SIM), STimulated Emission Depletion (STED) microscopy and Single Molecule Localisation Microscopy (SMLM) enable an increase in image resolution beyond the classical diffraction-limit. Here, we compare the individual strengths and weaknesses of each technique by imaging a variety of different subcellular structures in fixed cells. We chose examples ranging from well separated vesicles to densely packed three dimensional filaments. We used quantitative and correlative analyses to assess the performance of SIM, STED and SMLM with the aim of establishing a rough guideline regarding the suitability for typical applications and to highlight pitfalls associated with the different techniques.

  3. Compact three-dimensional super-resolution system based on fluorescence emission difference microscopy

    Science.gov (United States)

    Zhu, Dazhao; Chen, Youhua; Fang, Yue; Hussain, Anwar; Kuang, Cuifang; Zhou, Xiaoxu; Xu, Yingke; Liu, Xu

    2017-12-01

    A compact microscope system for three-dimensional (3-D) super-resolution imaging is presented. The super-resolution capability of the system is based on a size-reduced effective 3-D point spread function generated through the fluorescence emission difference (FED) method. The appropriate polarization direction distribution and manipulation allows the panel active area of the spatial light modulator to be fully utilized. This allows simultaneous modulation of the incident light by two kinds of phase masks to be performed with a single spatial light modulator in order to generate a 3-D negative spot. The system is more compact than standard 3-D FED systems while maintaining all the advantages of 3-D FED microscopy. The experimental results demonstrated the improvement in 3-D resolution by nearly 1.7 times and 1.6 times compared to the classic confocal resolution in the lateral and axial directions, respectively.

  4. Actin restructuring during Salmonella typhimurium infection investigated by confocal and super-resolution microscopy

    Science.gov (United States)

    Han, Jason J.; Kunde, Yuliya A.; Hong-Geller, Elizabeth; Werner, James H.

    2014-01-01

    We have used super-resolution optical microscopy and confocal microscopy to visualize the cytoskeletal restructuring of HeLa cells that accompanies and enables Salmonella typhimurium internalization. Herein, we report the use of confocal microscopy to verify and explore infection conditions that would be compatible with super-resolution optical microscopy, using Alexa-488 labeled phalloidin to stain the actin cytoskeletal network. While it is well known that actin restructuring and cytoskeletal rearrangements often accompany and assist in bacterial infection, most studies have employed conventional diffraction-limited fluorescence microscopy to explore these changes. Here we show that the superior spatial resolution provided by single-molecule localization methods (such as direct stochastic optical reconstruction microscopy) enables more precise visualization of the nanoscale changes in the actin cytoskeleton that accompany bacterial infection. In particular, we found that a thin (100-nm) ring of actin often surrounds an invading bacteria 10 to 20 min postinfection, with this ring being transitory in nature. We estimate that a few hundred monofilaments of actin surround the S. typhimurium in this heretofore unreported bacterial internalization intermediate.

  5. Super-resolution and super-localization microscopy: A novel tool for imaging chemical and biological processes

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Bin [Iowa State Univ., Ames, IA (United States)

    2015-01-01

    Optical microscopy imaging of single molecules and single particles is an essential method for studying fundamental biological and chemical processes at the molecular and nanometer scale. The best spatial resolution (~ λ/2) achievable in traditional optical microscopy is governed by the diffraction of light. However, single molecule-based super-localization and super-resolution microscopy imaging techniques have emerged in the past decade. Individual molecules can be localized with nanometer scale accuracy and precision for studying of biological and chemical processes.This work uncovered the heterogeneous properties of the pore structures. In this dissertation, the coupling of molecular transport and catalytic reaction at the single molecule and single particle level in multilayer mesoporous nanocatalysts was elucidated. Most previous studies dealt with these two important phenomena separately. A fluorogenic oxidation reaction of non-fluorescent amplex red to highly fluorescent resorufin was tested. The diffusion behavior of single resorufin molecules in aligned nanopores was studied using total internal reflection fluorescence microscopy (TIRFM).

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

    Science.gov (United States)

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

    2013-01-01

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

  7. 3D single-molecule super-resolution microscopy with a tilted light sheet.

    Science.gov (United States)

    Gustavsson, Anna-Karin; Petrov, Petar N; Lee, Maurice Y; Shechtman, Yoav; Moerner, W E

    2018-01-09

    Tilted light sheet microscopy with 3D point spread functions (TILT3D) combines a novel, tilted light sheet illumination strategy with long axial range point spread functions (PSFs) for low-background, 3D super-localization of single molecules as well as 3D super-resolution imaging in thick cells. Because the axial positions of the single emitters are encoded in the shape of each single-molecule image rather than in the position or thickness of the light sheet, the light sheet need not be extremely thin. TILT3D is built upon a standard inverted microscope and has minimal custom parts. The result is simple and flexible 3D super-resolution imaging with tens of nm localization precision throughout thick mammalian cells. We validate TILT3D for 3D super-resolution imaging in mammalian cells by imaging mitochondria and the full nuclear lamina using the double-helix PSF for single-molecule detection and the recently developed tetrapod PSFs for fiducial bead tracking and live axial drift correction.

  8. Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin; Lohse, Samuel E.; Lee, Chang-Soo; Torelli, Marco; Hamers, Robert J.; Murphy, Catherine; Orr, Galya; Haynes, Christy L.

    2014-01-01

    A method to fluorescently stain the surfaces of both Gram-negative and Gram-positive bacterial cells compatible with super-resolution fluorescence microscopy is presented. This method utilizes a commercially-available fluorescent probe to label primary amines at the surface of the cell. We demonstrate efficient staining of two bacterial strains, the Gram-negative Shewanella oneidensis MR-1 and the Gram-positive Bacillus subtilis 168. Using structured illumination microscopy and stochastic optical reconstruction microscopy, which require high quantum yield or specialized dyes, we show that this staining method may be used to resolve the bacterial cell surface with sub-diffraction-limited resolution. We further use this method to identify localization patterns of nanomaterials, specifically cadmium selenide quantum dots, following interaction with bacterial cells.

  9. Managing the Introduction of Super-Resolution Microscopy into a Core Facility.

    Science.gov (United States)

    Kamykowski, Jeffrey A; Storrie, Brian

    2017-01-01

    Super resolution techniques place the resolution of fluorescence microscopy closer to the size of the underlying cell structure or molecular machine being studied. Structured illumination techniques will give users a set of tools that are close to their past experience and relatively simple and quick to learn. The present dyes can be used. Resolution approaching 100 nm XY can be achieved. In contrast, stochastic methods such as PALM/STORM typically require the choice of new dyes and a much greater learning curve to master the technology and calculations. However, a further fivefold resolution improvement is possible. Stimulated depletion techniques such as STED offer a third set of approaches that will again require the use of new dyes. All these approaches require substantial investment in new equipment and in user training. There is no free lunch in the search for better resolution.

  10. Super-resolution imaging of subcortical white matter using stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI)

    Science.gov (United States)

    Hainsworth, A. H.; Lee, S.; Patel, A.; Poon, W. W.; Knight, A. E.

    2018-01-01

    Aims The spatial resolution of light microscopy is limited by the wavelength of visible light (the ‘diffraction limit’, approximately 250 nm). Resolution of sub-cellular structures, smaller than this limit, is possible with super resolution methods such as stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI). We aimed to resolve subcellular structures (axons, myelin sheaths and astrocytic processes) within intact white matter, using STORM and SOFI. Methods Standard cryostat-cut sections of subcortical white matter from donated human brain tissue and from adult rat and mouse brain were labelled, using standard immunohistochemical markers (neurofilament-H, myelin-associated glycoprotein, glial fibrillary acidic protein, GFAP). Image sequences were processed for STORM (effective pixel size 8–32 nm) and for SOFI (effective pixel size 80 nm). Results In human, rat and mouse, subcortical white matter high-quality images for axonal neurofilaments, myelin sheaths and filamentous astrocytic processes were obtained. In quantitative measurements, STORM consistently underestimated width of axons and astrocyte processes (compared with electron microscopy measurements). SOFI provided more accurate width measurements, though with somewhat lower spatial resolution than STORM. Conclusions Super resolution imaging of intact cryo-cut human brain tissue is feasible. For quantitation, STORM can under-estimate diameters of thin fluorescent objects. SOFI is more robust. The greatest limitation for super-resolution imaging in brain sections is imposed by sample preparation. We anticipate that improved strategies to reduce autofluorescence and to enhance fluorophore performance will enable rapid expansion of this approach. PMID:28696566

  11. Super-resolution imaging of subcortical white matter using stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI).

    Science.gov (United States)

    Hainsworth, A H; Lee, S; Foot, P; Patel, A; Poon, W W; Knight, A E

    2017-07-11

    The spatial resolution of light microscopy is limited by the wavelength of visible light (the 'diffraction limit', approximately 250 nm). Resolution of sub-cellular structures, smaller than this limit, is possible with super resolution methods such as stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI). We aimed to resolve subcellular structures (axons, myelin sheaths and astrocytic processes) within intact white matter, using STORM and SOFI. Standard cryostat-cut sections of subcortical white matter from donated human brain tissue and from adult rat and mouse brain were labelled, using standard immunohistochemical markers (neurofilament-H, myelin-associated glycoprotein, glial fibrillary acidic protein, GFAP). Image sequences were processed for STORM (effective pixel size 8-32 nm) and for SOFI (effective pixel size 80 nm). In human, rat and mouse, subcortical white matter high-quality images for axonal neurofilaments, myelin sheaths and filamentous astrocytic processes were obtained. In quantitative measurements, STORM consistently underestimated width of axons and astrocyte processes (compared with electron microscopy measurements). SOFI provided more accurate width measurements, though with somewhat lower spatial resolution than STORM. Super resolution imaging of intact cryo-cut human brain tissue is feasible. For quantitation, STORM can under-estimate diameters of thin fluorescent objects. SOFI is more robust. The greatest limitation for super-resolution imaging in brain sections is imposed by sample preparation. We anticipate that improved strategies to reduce autofluorescence and to enhance fluorophore performance will enable rapid expansion of this approach. © 2017 British Neuropathological Society.

  12. Revealing t-tubules in striated muscle with new optical super-resolution microscopy techniques

    Directory of Open Access Journals (Sweden)

    Isuru D. Jayasinghe

    2014-12-01

    Full Text Available The t-tubular system plays a central role in the synchronisation of calcium signalling and excitation-contraction coupling in most striated muscle cells. Light microscopy has been used for imaging t-tubules for well over 100 years and together with electron microscopy (EM, has revealed the three-dimensional complexities of the t-system topology within cardiomyocytes and skeletal muscle fibres from a range of species. The emerging super-resolution single molecule localisation microscopy (SMLM techniques are offering a near 10-fold improvement over the resolution of conventional fluorescence light microscopy methods, with the ability to spectrally resolve nanometre scale distributions of multiple molecular targets. In conjunction with the next generation of electron microscopy, SMLM has allowed the visualisation and quantification of intricate t-tubule morphologies within large areas of muscle cells at an unprecedented level of detail. In this paper, we review recent advancements in the t-tubule structural biology with the utility of various microscopy techniques. We outline the technical considerations in adapting SMLM to study t-tubules and its potential to further our understanding of the molecular processes that underlie the sub-micron scale structural alterations observed in a range of muscle pathologies.

  13. CINCH (confocal incoherent correlation holography) super resolution fluorescence microscopy based upon FINCH (Fresnel incoherent correlation holography).

    Science.gov (United States)

    Siegel, Nisan; Storrie, Brian; Bruce, Marc; Brooker, Gary

    2015-02-07

    FINCH holographic fluorescence microscopy creates high resolution super-resolved images with enhanced depth of focus. The simple addition of a real-time Nipkow disk confocal image scanner in a conjugate plane of this incoherent holographic system is shown to reduce the depth of focus, and the combination of both techniques provides a simple way to enhance the axial resolution of FINCH in a combined method called "CINCH". An important feature of the combined system allows for the simultaneous real-time image capture of widefield and holographic images or confocal and confocal holographic images for ready comparison of each method on the exact same field of view. Additional GPU based complex deconvolution processing of the images further enhances resolution.

  14. HIV taken by STORM: Super-resolution fluorescence microscopy of a viral infection

    Directory of Open Access Journals (Sweden)

    Pereira Cândida F

    2012-05-01

    Full Text Available Abstract Background The visualization of viral proteins has been hindered by the resolution limit of conventional fluorescent microscopes, as the dimension of any single fluorescent signal is often greater than most virion particles. Super-resolution microscopy has the potential to unveil the distribution of proteins at the resolution approaching electron microscopy without relying on morphological features of existing characteristics of the biological specimen that are needed in EM. Results Using direct stochastic optical reconstruction microscopy (dSTORM to achieve a lateral resolution of 15–20 nm, we quantified the 2-D molecular distribution of the major structural proteins of the infectious human immunodeficiency virus type 1 (HIV-1 before and after infection of lymphoid cells. We determined that the HIV-1 matrix and capsid proteins undergo restructuring soon after HIV-1 infection. Conclusions This study provides the proof-of-concept for the use of dSTORM to visualize the changes in the molecular distribution of viral proteins during an infection.

  15. Super-resolution microscopy as a potential approach to diagnosis of platelet granule disorders.

    Science.gov (United States)

    Westmoreland, D; Shaw, M; Grimes, W; Metcalf, D J; Burden, J J; Gomez, K; Knight, A E; Cutler, D F

    2016-04-01

    Many platelet functions are dependent on bioactive molecules released from their granules. Deficiencies of these granules in number, shape or content are associated with bleeding. The small size of these granules is such that imaging them for diagnosis has traditionally required electron microscopy. However, recently developed super-resolution microscopes provide sufficient spatial resolution to effectively image platelet granules. When combined with automated image analysis, these methods provide a quantitative, unbiased, rapidly acquired dataset that can readily and reliably reveal differences in platelet granules between individuals. To demonstrate the ability of structured illumination microscopy (SIM) to efficiently differentiate between healthy volunteers and three patients with Hermansky-Pudlak syndrome. Blood samples were taken from three patients with Hermansky-Pudlak syndrome and seven controls. Patients 1-3 have gene defects in HPS1, HPS6 and HPS5, respectively; all controls were healthy volunteers. Platelet-rich plasma was isolated from blood and the platelets fixed, stained for CD63 and processed for analysis by immunofluorescence microscopy, using a custom-built SIM microscope. SIM can successfully resolve CD63-positive structures in fixed platelets. A determination of the number of CD63-positive structures per platelet allowed us to conclude that each patient was significantly different from all of the controls with 99% confidence. A super-resolution imaging approach is effective and rapid in objectively differentiating between patients with a platelet bleeding disorder and healthy volunteers. CD63 is a useful marker for predicting Hermansky-Pudlak syndrome and could be used in the diagnosis of patients suspected of other platelet granule disorders. © 2016 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals, Inc. on behalf of International Society on Thrombosis and Haemostasis.

  16. Measurement of replication structures at the nanometer scale using super-resolution light microscopy.

    Science.gov (United States)

    Baddeley, D; Chagin, V O; Schermelleh, L; Martin, S; Pombo, A; Carlton, P M; Gahl, A; Domaing, P; Birk, U; Leonhardt, H; Cremer, C; Cardoso, M C

    2010-01-01

    DNA replication, similar to other cellular processes, occurs within dynamic macromolecular structures. Any comprehensive understanding ultimately requires quantitative data to establish and test models of genome duplication. We used two different super-resolution light microscopy techniques to directly measure and compare the size and numbers of replication foci in mammalian cells. This analysis showed that replication foci vary in size from 210 nm down to 40 nm. Remarkably, spatially modulated illumination (SMI) and 3D-structured illumination microscopy (3D-SIM) both showed an average size of 125 nm that was conserved throughout S-phase and independent of the labeling method, suggesting a basic unit of genome duplication. Interestingly, the improved optical 3D resolution identified 3- to 5-fold more distinct replication foci than previously reported. These results show that optical nanoscopy techniques enable accurate measurements of cellular structures at a level previously achieved only by electron microscopy and highlight the possibility of high-throughput, multispectral 3D analyses.

  17. Painting Supramolecular Polymers in Organic Solvents by Super-resolution Microscopy

    Science.gov (United States)

    2018-01-01

    Despite the rapid development of complex functional supramolecular systems, visualization of these architectures under native conditions at high resolution has remained a challenging endeavor. Super-resolution microscopy was recently proposed as an effective tool to unveil one-dimensional nanoscale structures in aqueous media upon chemical functionalization with suitable fluorescent probes. Building upon our previous work, which enabled photoactivation localization microscopy in organic solvents, herein, we present the imaging of one-dimensional supramolecular polymers in their native environment by interface point accumulation for imaging in nanoscale topography (iPAINT). The noncovalent staining, typical of iPAINT, allows the investigation of supramolecular polymers’ structure in situ without any chemical modification. The quasi-permanent adsorption of the dye to the polymer is exploited to identify block-like arrangements within supramolecular fibers, which were obtained upon mixing homopolymers that were prestained with different colors. The staining of the blocks, maintained by the lack of exchange of the dyes, permits the imaging of complex structures for multiple days. This study showcases the potential of PAINT-like strategies such as iPAINT to visualize multicomponent dynamic systems in their native environment with an easy, synthesis-free approach and high spatial resolution. PMID:29697958

  18. Brain Slice Staining and Preparation for Three-Dimensional Super-Resolution Microscopy

    Science.gov (United States)

    German, Christopher L.; Gudheti, Manasa V.; Fleckenstein, Annette E.; Jorgensen, Erik M.

    2018-01-01

    Localization microscopy techniques – such as photoactivation localization microscopy (PALM), fluorescent PALM (FPALM), ground state depletion (GSD), and stochastic optical reconstruction microscopy (STORM) – provide the highest precision for single molecule localization currently available. However, localization microscopy has been largely limited to cell cultures due to the difficulties that arise in imaging thicker tissue sections. Sample fixation and antibody staining, background fluorescence, fluorophore photoinstability, light scattering in thick sections, and sample movement create significant challenges for imaging intact tissue. We have developed a sample preparation and image acquisition protocol to address these challenges in rat brain slices. The sample preparation combined multiple fixation steps, saponin permeabilization, and tissue clarification. Together, these preserve intracellular structures, promote antibody penetration, reduce background fluorescence and light scattering, and allow acquisition of images deep in a 30 μm thick slice. Image acquisition challenges were resolved by overlaying samples with a permeable agarose pad and custom-built stainless steel imaging adapter, and sealing the imaging chamber. This approach kept slices flat, immobile, bathed in imaging buffer, and prevented buffer oxidation during imaging. Using this protocol, we consistently obtained single molecule localizations of synaptic vesicle and active zone proteins in three-dimensions within individual synaptic terminals of the striatum in rat brain slices. These techniques may be easily adapted to the preparation and imaging of other tissues, substantially broadening the application of super-resolution imaging. PMID:28924666

  19. Synthesis of a Far-Red Photoactivatable Silicon-Containing Rhodamine for Super-Resolution Microscopy.

    Science.gov (United States)

    Grimm, Jonathan B; Klein, Teresa; Kopek, Benjamin G; Shtengel, Gleb; Hess, Harald F; Sauer, Markus; Lavis, Luke D

    2016-01-26

    The rhodamine system is a flexible framework for building small-molecule fluorescent probes. Changing N-substitution patterns and replacing the xanthene oxygen with a dimethylsilicon moiety can shift the absorption and fluorescence emission maxima of rhodamine dyes to longer wavelengths. Acylation of the rhodamine nitrogen atoms forces the molecule to adopt a nonfluorescent lactone form, providing a convenient method to make fluorogenic compounds. Herein, we take advantage of all of these structural manipulations and describe a novel photoactivatable fluorophore based on a Si-containing analogue of Q-rhodamine. This probe is the first example of a "caged" Si-rhodamine, exhibits higher photon counts compared to established localization microscopy dyes, and is sufficiently red-shifted to allow multicolor imaging. The dye is a useful label for super-resolution imaging and constitutes a new scaffold for far-red fluorogenic molecules. © 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

  20. Combined multi-plane phase retrieval and super-resolution optical fluctuation imaging for 4D cell microscopy

    Science.gov (United States)

    Descloux, A.; Grußmayer, K. S.; Bostan, E.; Lukes, T.; Bouwens, A.; Sharipov, A.; Geissbuehler, S.; Mahul-Mellier, A.-L.; Lashuel, H. A.; Leutenegger, M.; Lasser, T.

    2018-03-01

    Super-resolution fluorescence microscopy provides unprecedented insight into cellular and subcellular structures. However, going `beyond the diffraction barrier' comes at a price, since most far-field super-resolution imaging techniques trade temporal for spatial super-resolution. We propose the combination of a novel label-free white light quantitative phase imaging with fluorescence to provide high-speed imaging and spatial super-resolution. The non-iterative phase retrieval relies on the acquisition of single images at each z-location and thus enables straightforward 3D phase imaging using a classical microscope. We realized multi-plane imaging using a customized prism for the simultaneous acquisition of eight planes. This allowed us to not only image live cells in 3D at up to 200 Hz, but also to integrate fluorescence super-resolution optical fluctuation imaging within the same optical instrument. The 4D microscope platform unifies the sensitivity and high temporal resolution of phase imaging with the specificity and high spatial resolution of fluorescence microscopy.

  1. Quantum Dot Immunocytochemical Localization of Somatostatin in Somatostatinoma by Widefield Epifluorescence, Super-resolution Light, and Immunoelectron Microscopy

    Science.gov (United States)

    Lai, Ken; Wu, Xiaojuan; Yong, Jim L. C.; Lee, C. Soon

    2012-01-01

    Quantum dot nanocrystal probes (QDs) have been used for detection of somatostatin hormone in secretory granules of somatostatinoma tumor cells by immunofluorescence light microscopy, super-resolution light microscopy, and immunoelectron microscopy. Immunostaining for all modalities was done using sections taken from an epoxy resin-embedded tissue specimen and a similar labeling protocol. This approach allowed assessment of labeling at light microscopy level before examination at super-resolution and electron microscopy level and was a significant aid in interpretation. Etching of ultrathin sections with saturated sodium metaperiodate was a critical step presumably able to retrieve some tissue antigenicity masked by processing in epoxy resin. Immunofluorescence microscopy of QD-immunolabeled sections showed somatostatin hormone localization in cytoplasmic granules. Some variable staining of tumor gland-like structures appeared related to granule maturity and dispersal of granule contents within the tumor cell cytoplasm. Super-resolution light microscopy demonstrated localization of somatostatin within individual secretory granules to be heterogeneous, and this staining pattern was confirmed by immunoelectron microscopy. PMID:22899862

  2. Quantum dot immunocytochemical localization of somatostatin in somatostatinoma by Widefield Epifluorescence, super-resolution light, and immunoelectron microscopy.

    Science.gov (United States)

    Killingsworth, Murray C; Lai, Ken; Wu, Xiaojuan; Yong, Jim L C; Lee, C Soon

    2012-11-01

    Quantum dot nanocrystal probes (QDs) have been used for detection of somatostatin hormone in secretory granules of somatostatinoma tumor cells by immunofluorescence light microscopy, super-resolution light microscopy, and immunoelectron microscopy. Immunostaining for all modalities was done using sections taken from an epoxy resin-embedded tissue specimen and a similar labeling protocol. This approach allowed assessment of labeling at light microscopy level before examination at super-resolution and electron microscopy level and was a significant aid in interpretation. Etching of ultrathin sections with saturated sodium metaperiodate was a critical step presumably able to retrieve some tissue antigenicity masked by processing in epoxy resin. Immunofluorescence microscopy of QD-immunolabeled sections showed somatostatin hormone localization in cytoplasmic granules. Some variable staining of tumor gland-like structures appeared related to granule maturity and dispersal of granule contents within the tumor cell cytoplasm. Super-resolution light microscopy demonstrated localization of somatostatin within individual secretory granules to be heterogeneous, and this staining pattern was confirmed by immunoelectron microscopy.

  3. Enhanced simulator software for image validation and interpretation for multimodal localization super-resolution fluorescence microscopy

    Science.gov (United States)

    Erdélyi, Miklós; Sinkó, József; Gajdos, Tamás.; Novák, Tibor

    2017-02-01

    Optical super-resolution techniques such as single molecule localization have become one of the most dynamically developed areas in optical microscopy. These techniques routinely provide images of fixed cells or tissues with sub-diffraction spatial resolution, and can even be applied for live cell imaging under appropriate circumstances. Localization techniques are based on the precise fitting of the point spread functions (PSF) to the measured images of stochastically excited, identical fluorescent molecules. These techniques require controlling the rate between the on, off and the bleached states, keeping the number of active fluorescent molecules at an optimum value, so their diffraction limited images can be detected separately both spatially and temporally. Because of the numerous (and sometimes unknown) parameters, the imaging system can only be handled stochastically. For example, the rotation of the dye molecules obscures the polarization dependent PSF shape, and only an averaged distribution - typically estimated by a Gaussian function - is observed. TestSTORM software was developed to generate image stacks for traditional localization microscopes, where localization meant the precise determination of the spatial position of the molecules. However, additional optical properties (polarization, spectra, etc.) of the emitted photons can be used for further monitoring the chemical and physical properties (viscosity, pH, etc.) of the local environment. The image stack generating program was upgraded by several new features, such as: multicolour, polarization dependent PSF, built-in 3D visualization, structured background. These features make the program an ideal tool for optimizing the imaging and sample preparation conditions.

  4. Current limitations in super-resolution fluorescence microscopy for biological specimens: How deep can we go from the cover glass?

    Science.gov (United States)

    Okada, Yasushi

    2017-04-01

    Diffraction limit of resolution has been one of the biggest limitations in the optical microscopy. Super-resolution fluorescence microscopy has enabled us to break this limit. However, for the observations of real biological specimens, especially for the imaging of tissues or whole body, the target structures of interest are often embedded deep inside the specimen. Here, we would present our results to extend the target of the super-resolution microscopy deeper into the cells. Confocal microscope optics work effectively to minimize the effect by the aberrations by the cellular components, but at the expense of the signal intensities. Spherical aberrations by the refractive index mismatch between the cellular environment and the immersion liquid can be much larger, but can be reduced by adjusting the correction collar at the objective lens.

  5. Super-Resolution Microscopy Reveals the Native Ultrastructure of the Erythrocyte Cytoskeleton

    Directory of Open Access Journals (Sweden)

    Leiting Pan

    2018-01-01

    Full Text Available The erythrocyte cytoskeleton is a textbook prototype for the submembrane cytoskeleton of metazoan cells. While early experiments suggest a triangular network of actin-based junctional complexes connected by ∼200-nm-long spectrin tetramers, later studies indicate much smaller junction-to-junction distances in the range of 25-60 nm. Through super-resolution microscopy, we resolve the native ultrastructure of the cytoskeleton of membrane-preserved erythrocytes for the N and C termini of β-spectrin, F-actin, protein 4.1, tropomodulin, and adducin. This allows us to determine an ∼80-nm junction-to-junction distance, a length consistent with relaxed spectrin tetramers and theories based on spectrin abundance. Through two-color data, we further show that the cytoskeleton meshwork often contains nanoscale voids where the cell membrane remains intact and that actin filaments and capping proteins localize to a subset of, but not all, junctional complexes. Together, our results call for a reassessment of the structure and function of the submembrane cytoskeleton.

  6. Super-resolution optical microscopy resolves network morphology of smart colloidal microgels.

    Science.gov (United States)

    Bergmann, Stephan; Wrede, Oliver; Huser, Thomas; Hellweg, Thomas

    2018-02-14

    We present a new method to resolve the network morphology of colloidal particles in an aqueous environment via super-resolution microscopy. By localization of freely diffusing fluorophores inside the particle network we can resolve the three dimensional structure of one species of colloidal particles (thermoresponsive microgels) without altering their chemical composition through copolymerization with fluorescent monomers. Our approach utilizes the interaction of the fluorescent dye rhodamine 6G with the polymer network to achieve an indirect labeling. We calculate the 3D structure from the 2D images and compare the structure to previously published models for the microgel morphology, e.g. the fuzzy sphere model. To describe the differences in the data an extension of this model is suggested. Our method enables the tailor-made fabrication of colloidal particles which are used in various applications, such as paints or cosmetics, and are promising candidates for drug delivery, smart surface coatings, and nanocatalysis. With the precise knowledge of the particle morphology an understanding of the underlying structure-property relationships for various colloidal systems is possible.

  7. Super-resolution binding activated localization microscopy through reversible change of DNA conformation.

    Science.gov (United States)

    Szczurek, Aleksander; Birk, Udo; Knecht, Hans; Dobrucki, Jurek; Mai, Sabine; Cremer, Christoph

    2018-01-01

    Methods of super-resolving light microscopy (SRM) have found an exponentially growing range of applications in cell biology, including nuclear structure analyses. Recent developments have proven that Single Molecule Localization Microscopy (SMLM), a type of SRM, is particularly useful for enhanced spatial analysis of the cell nucleus due to its highest resolving capability combined with very specific fluorescent labeling. In this commentary we offer a brief review of the latest methodological development in the field of SMLM of chromatin designated DNA Structure Fluctuation Assisted Binding Activated Localization Microscopy (abbreviated as fBALM) as well as its potential future applications in biology and medicine.

  8. 4D super-resolution microscopy with conventional fluorophores and single wavelength excitation in optically thick cells and tissues.

    Directory of Open Access Journals (Sweden)

    David Baddeley

    Full Text Available BACKGROUND: Optical super-resolution imaging of fluorescently stained biological samples is rapidly becoming an important tool to investigate protein distribution at the molecular scale. It is therefore important to develop practical super-resolution methods that allow capturing the full three-dimensional nature of biological systems and also can visualize multiple protein species in the same sample. METHODOLOGY/PRINCIPAL FINDINGS: We show that the use of a combination of conventional near-infrared dyes, such as Alexa 647, Alexa 680 and Alexa 750, all excited with a 671 nm diode laser, enables 3D multi-colour super-resolution imaging of complex biological samples. Optically thick samples, including human tissue sections, cardiac rat myocytes and densely grown neuronal cultures were imaged with lateral resolutions of ∼15 nm (std. dev. while reducing marker cross-talk to <1%. Using astigmatism an axial resolution of ∼65 nm (std. dev. was routinely achieved. The number of marker species that can be distinguished depends on the mean photon number of single molecule events. With the typical photon yields from Alexa 680 of ∼2000 up to 5 markers may in principle be resolved with <2% crosstalk. CONCLUSIONS/SIGNIFICANCE: Our approach is based entirely on the use of conventional, commercially available markers and requires only a single laser. It provides a very straightforward way to investigate biological samples at the nanometre scale and should help establish practical 4D super-resolution microscopy as a routine research tool in many laboratories.

  9. Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures

    International Nuclear Information System (INIS)

    Reuss, Matthias; Blom, Hans; Brismar, Hjalmar; Fördős, Ferenc; Högberg, Björn; Öktem, Ozan

    2017-01-01

    A common method to assess the performance of (super resolution) microscopes is to use the localization precision of emitters as an estimate for the achieved resolution. Naturally, this is widely used in super resolution methods based on single molecule stochastic switching. This concept suffers from the fact that it is hard to calibrate measures against a real sample (a phantom), because true absolute positions of emitters are almost always unknown. For this reason, resolution estimates are potentially biased in an image since one is blind to true position accuracy, i.e. deviation in position measurement from true positions. We have solved this issue by imaging nanorods fabricated with DNA-origami. The nanorods used are designed to have emitters attached at each end in a well-defined and highly conserved distance. These structures are widely used to gauge localization precision. Here, we additionally determined the true achievable localization accuracy and compared this figure of merit to localization precision values for two common super resolution microscope methods STED and STORM. (paper)

  10. Spectrally Resolved and Functional Super-resolution Microscopy via Ultrahigh-Throughput Single-Molecule Spectroscopy.

    Science.gov (United States)

    Yan, Rui; Moon, Seonah; Kenny, Samuel J; Xu, Ke

    2018-03-20

    As an elegant integration of the spatial and temporal dimensions of single-molecule fluorescence, single-molecule localization microscopy (SMLM) overcomes the diffraction-limited resolution barrier of optical microscopy by localizing single molecules that stochastically switch between fluorescent and dark states over time. While this type of super-resolution microscopy (SRM) technique readily achieves remarkable spatial resolutions of ∼10 nm, it typically provides no spectral information. Meanwhile, current scanning-based single-location approaches for mapping the positions and spectra of single molecules are limited by low throughput and are difficult to apply to densely labeled (bio)samples. In this Account, we summarize the rationale, design, and results of our recent efforts toward the integration of the spectral dimension of single-molecule fluorescence with SMLM to achieve spectrally resolved SMLM (SR-SMLM) and functional SRM ( f-SRM). By developing a wide-field scheme for spectral measurement and implementing single-molecule fluorescence on-off switching typical of SMLM, we first showed that in densely labeled (bio)samples it is possible to record the fluorescence spectra and positions of millions of single molecules synchronously within minutes, giving rise to ultrahigh-throughput single-molecule spectroscopy and SR-SMLM. This allowed us to first show statistically that for many dyes, single molecules of the same species exhibit near identical emission in fixed cells. This narrow distribution of emission wavelengths, which contrasts markedly with previous results at solid surfaces, allowed us to unambiguously identify single molecules of spectrally similar dyes. Crosstalk-free, multiplexed SRM was thus achieved for four dyes that were merely 10 nm apart in emission spectrum, with the three-dimensional SRM images of all four dyes being automatically aligned within one image channel. The ability to incorporate single-molecule fluorescence measurement with

  11. Reconstitution radicicol containing apolipoprotein B lipoparticle and tracing its cell uptake process by super resolution fluorescent microscopy.

    Science.gov (United States)

    Lin, Chung Ching; Lin, Po-Yen; Chang, Chia-Ching

    Apolipoprotein B (apoB) is the only protein of LDL. LDL delivers cholesterol, triacylglycerides and lipids to the target cells. Reconstitute apoB lipoparticle (rABL) will be an idea drug delivery vehicle for hydrophobic and amphiphilic materials delivery. It is challenged to renature ApoB into its functional state from denatured state. By using modified bile salt and radicicol (Rad) added over-critical refolding process, apoB can be restored into its native like state. The intrinsic fluorescence of apoB increased during the refolding process. Moreover, radicicol (Rad) molecules have been encapsulated into reconstitute rABL (Rad@rABL). To investigate the cell uptake mechanism of Rad@rABL, a super resolution ground state depletion (GSD) microscopy is used in this research. Fluorescence labeled Rad@rABL can be traced within the tumor cell. Key words: LDL, radicicol, protein refolding, super resolution microscopy.

  12. Challenges for Super-Resolution Localization Microscopy and Biomolecular Fluorescent Nano-Probing in Cancer Research

    Science.gov (United States)

    Ilić, Nataša; Pilarczyk, Götz; Lee, Jin-Ho; Logeswaran, Abiramy; Borroni, Aurora Paola; Krufczik, Matthias; Theda, Franziska; Waltrich, Nadine; Bestvater, Felix; Hildenbrand, Georg; Cremer, Christoph; Blank, Michael

    2017-01-01

    Understanding molecular interactions and regulatory mechanisms in tumor initiation, progression, and treatment response are key requirements towards advanced cancer diagnosis and novel treatment procedures in personalized medicine. Beyond decoding the gene expression, malfunctioning and cancer-related epigenetic pathways, investigations of the spatial receptor arrangements in membranes and genome organization in cell nuclei, on the nano-scale, contribute to elucidating complex molecular mechanisms in cells and tissues. By these means, the correlation between cell function and spatial organization of molecules or molecular complexes can be studied, with respect to carcinogenesis, tumor sensitivity or tumor resistance to anticancer therapies, like radiation or antibody treatment. Here, we present several new applications for bio-molecular nano-probes and super-resolution, laser fluorescence localization microscopy and their potential in life sciences, especially in biomedical and cancer research. By means of a tool-box of fluorescent antibodies, green fluorescent protein (GFP) tagging, or specific oligonucleotides, we present tumor relevant re-arrangements of Erb-receptors in membranes, spatial organization of Smad specific ubiquitin protein ligase 2 (Smurf2) in the cytosol, tumor cell characteristic heterochromatin organization, and molecular re-arrangements induced by radiation or antibody treatment. The main purpose of this article is to demonstrate how nano-scaled distance measurements between bio-molecules, tagged by appropriate nano-probes, can be applied to elucidate structures and conformations of molecular complexes which are characteristic of tumorigenesis and treatment responses. These applications open new avenues towards a better interpretation of the spatial organization and treatment responses of functionally relevant molecules, at the single cell level, in normal and cancer cells, offering new potentials for individualized medicine. PMID:28956810

  13. Challenges for Super-Resolution Localization Microscopy and Biomolecular Fluorescent Nano-Probing in Cancer Research

    Directory of Open Access Journals (Sweden)

    Michael Hausmann

    2017-09-01

    Full Text Available Understanding molecular interactions and regulatory mechanisms in tumor initiation, progression, and treatment response are key requirements towards advanced cancer diagnosis and novel treatment procedures in personalized medicine. Beyond decoding the gene expression, malfunctioning and cancer-related epigenetic pathways, investigations of the spatial receptor arrangements in membranes and genome organization in cell nuclei, on the nano-scale, contribute to elucidating complex molecular mechanisms in cells and tissues. By these means, the correlation between cell function and spatial organization of molecules or molecular complexes can be studied, with respect to carcinogenesis, tumor sensitivity or tumor resistance to anticancer therapies, like radiation or antibody treatment. Here, we present several new applications for bio-molecular nano-probes and super-resolution, laser fluorescence localization microscopy and their potential in life sciences, especially in biomedical and cancer research. By means of a tool-box of fluorescent antibodies, green fluorescent protein (GFP tagging, or specific oligonucleotides, we present tumor relevant re-arrangements of Erb-receptors in membranes, spatial organization of Smad specific ubiquitin protein ligase 2 (Smurf2 in the cytosol, tumor cell characteristic heterochromatin organization, and molecular re-arrangements induced by radiation or antibody treatment. The main purpose of this article is to demonstrate how nano-scaled distance measurements between bio-molecules, tagged by appropriate nano-probes, can be applied to elucidate structures and conformations of molecular complexes which are characteristic of tumorigenesis and treatment responses. These applications open new avenues towards a better interpretation of the spatial organization and treatment responses of functionally relevant molecules, at the single cell level, in normal and cancer cells, offering new potentials for individualized medicine.

  14. Under the Microscope: Single-Domain Antibodies for Live-Cell Imaging and Super-Resolution Microscopy

    OpenAIRE

    Traenkle, Bjoern; Rothbauer, Ulrich

    2017-01-01

    Single-domain antibodies (sdAbs) have substantially expanded the possibilities of advanced cellular imaging such as live-cell or super-resolution microscopy to visualize cellular antigens and their dynamics. In addition to their unique properties including small size, high stability, and solubility in many environments, sdAbs can be efficiently functionalized according to the needs of the respective imaging approach. Genetically encoded intrabodies fused to fluorescent proteins (chromobodies)...

  15. Under the Microscope: Single-Domain Antibodies for Live-Cell Imaging and Super-Resolution Microscopy

    Directory of Open Access Journals (Sweden)

    Bjoern Traenkle

    2017-08-01

    Full Text Available Single-domain antibodies (sdAbs have substantially expanded the possibilities of advanced cellular imaging such as live-cell or super-resolution microscopy to visualize cellular antigens and their dynamics. In addition to their unique properties including small size, high stability, and solubility in many environments, sdAbs can be efficiently functionalized according to the needs of the respective imaging approach. Genetically encoded intrabodies fused to fluorescent proteins (chromobodies have become versatile tools to study dynamics of endogenous proteins in living cells. Additionally, sdAbs conjugated to organic dyes were shown to label cellular structures with high density and minimal fluorophore displacement making them highly attractive probes for super-resolution microscopy. Here, we review recent advances of the chromobody technology to visualize localization and dynamics of cellular targets and the application of chromobody-based cell models for compound screening. Acknowledging the emerging importance of super-resolution microscopy in cell biology, we further discuss advantages and challenges of sdAbs for this technology.

  16. Under the Microscope: Single-Domain Antibodies for Live-Cell Imaging and Super-Resolution Microscopy.

    Science.gov (United States)

    Traenkle, Bjoern; Rothbauer, Ulrich

    2017-01-01

    Single-domain antibodies (sdAbs) have substantially expanded the possibilities of advanced cellular imaging such as live-cell or super-resolution microscopy to visualize cellular antigens and their dynamics. In addition to their unique properties including small size, high stability, and solubility in many environments, sdAbs can be efficiently functionalized according to the needs of the respective imaging approach. Genetically encoded intrabodies fused to fluorescent proteins (chromobodies) have become versatile tools to study dynamics of endogenous proteins in living cells. Additionally, sdAbs conjugated to organic dyes were shown to label cellular structures with high density and minimal fluorophore displacement making them highly attractive probes for super-resolution microscopy. Here, we review recent advances of the chromobody technology to visualize localization and dynamics of cellular targets and the application of chromobody-based cell models for compound screening. Acknowledging the emerging importance of super-resolution microscopy in cell biology, we further discuss advantages and challenges of sdAbs for this technology.

  17. Contrast enhancement of microsphere-assisted super-resolution imaging in dark-field microscopy

    Science.gov (United States)

    Zhou, Yi; Tang, Yan; Deng, Qinyuan; Zhao, Lixin; Hu, Song

    2017-08-01

    We report a method of boosting the imaging contrast of microsphere-assisted super-resolution visualization by utilizing dark-field illumination (DFI). We conducted experiments on both 10-µm-diameter silica (SiO2) microspheres with refractive index n ∼ 1.46 under no and partial immersion in ethyl alcohol (n ∼ 1.36) and 20-µm-diameter barium titanate glass (BTG, n ∼ 1.9) microspheres with full immersion to show the super-resolution capability. We experimentally demonstrated that the imaging contrast and uniformity were extraordinarily improved in the DFI mode. The intensity profiles in the visualization also numerically confirm the enhanced sharpness for a better imaging quality when applying DFI.

  18. Super-Resolution Localization Microscopy of γ-H2AX and Heterochromatin after Folate Deficiency.

    Science.gov (United States)

    Bach, Margund; Savini, Claudia; Krufczik, Matthias; Cremer, Christoph; Rösl, Frank; Hausmann, Michael

    2017-08-08

    Folate is an essential water-soluble vitamin in food and nutrition supplements. As a one-carbon source, it is involved in many central regulatory processes, such as DNA, RNA, and protein methylation as well as DNA synthesis and repair. Deficiency in folate is considered to be associated with an increased incidence of several malignancies, including cervical cancer that is etiologically linked to an infection with "high-risk" human papilloma viruses (HPV). However, it is still not known how a recommended increase in dietary folate after its deprivation affects the physiological status of cells. To study the impact of folate depletion and its subsequent reconstitution in single cells, we used quantitative chromatin conformation measurements obtained by super-resolution fluorescence microscopy, i.e., single molecule localization microscopy (SMLM). As a read-out, we examined the levels and the (re)positioning of γ-H2AX tags and histone H3K9me3 heterochromatin tags after immunostaining in three-dimensional (3D)-conserved cell nuclei. As model, we used HPV16 positive immortalized human keratinocytes that were cultivated under normal, folate deficient, and reconstituted conditions for different periods of time. The results were compared to cells continuously cultivated in standard folate medium. After 13 weeks in low folate, an increase in the phosphorylation of the histone H2AX was noted, indicative of an accumulation of DNA double strand breaks. DNA repair activity represented by the formation of those γ-H2AX clusters was maintained during the following 15 weeks of examination. However, the clustered arrangements of tags appeared to relax in a time-dependent manner. Parallel to the repair activity, the chromatin methylation activity increased as detected by H3K9me3 tags. The progress of DNA double strand repair was accompanied by a reduction of the detected nucleosome density around the γ-H2AX clusters, suggesting a shift from hetero- to euchromatin to allow access

  19. Super-resolution Phase Tomography

    KAUST Repository

    Depeursinge, Christian; Cotte, Yann; Toy, Fatih; Jourdain, Pascal; Boss, Daiel; Marquet, Pierre; Magistretti, Pierre J.

    2013-01-01

    Digital Holographic Microscopy (DHM) yields reconstructed complex wavefields. It allows synthesizing the aperture of a virtual microscope up to 2π, offering super-resolution phase images. Live images of micro-organisms and neurons with resolution less than 100 nm are presented.

  20. Super-resolution Phase Tomography

    KAUST Repository

    Depeursinge, Christian

    2013-04-21

    Digital Holographic Microscopy (DHM) yields reconstructed complex wavefields. It allows synthesizing the aperture of a virtual microscope up to 2π, offering super-resolution phase images. Live images of micro-organisms and neurons with resolution less than 100 nm are presented.

  1. Parallel detecting super-resolution microscopy using correlation based image restoration

    Science.gov (United States)

    Yu, Zhongzhi; Liu, Shaocong; Zhu, Dazhao; Kuang, Cuifang; Liu, Xu

    2017-12-01

    A novel approach to achieve the image restoration is proposed in which each detector's relative position in the detector array is no longer a necessity. We can identify each detector's relative location by extracting a certain area from one of the detector's image and scanning it on other detectors' images. According to this location, we can generate the point spread functions (PSF) for each detector and perform deconvolution for image restoration. Equipped with this method, the microscope with discretionally designed detector array can be easily constructed without the concern of exact relative locations of detectors. The simulated results and experimental results show the total improvement in resolution with a factor of 1.7 compared to conventional confocal fluorescence microscopy. With the significant enhancement in resolution and easiness for application of this method, this novel method should have potential for a wide range of application in fluorescence microscopy based on parallel detecting.

  2. Structured Illumination-Based Super-Resolution Optical Microscopy for Hemato- and Cyto-Pathology Applications

    Directory of Open Access Journals (Sweden)

    Tieqiao Zhang

    2013-01-01

    Full Text Available Structured illumination fluorescence microscopy utilizes interfering light and the moiré effect to enhance spatial resolution to about a half of that of conventional light microscopy, i.e. approximately 90 nm. In addition to the enhancement in the x and y directions, it also allows enhancement of resolution in the z- direction by the same factor of two (to approximately 220 nm, making it a powerful tool for 3-D morphology studies of fluorescently labeled cells or thin tissue sections. In this report, we applied this technique to several types of blood cells that are commonly seen in hematopathology. Compared with standard brightfield and ordinary fluorescence microscopy images, the 3-D morphology results clearly reveal the morphological features of different types of normal blood cells. We have also used this technique to evaluate morphologies of abnormal erythrocytes and compare them with those recorded on normal cells. The results give a very intuitive presentation of morphological structures of erythrocytes with great details. This research illustrates the potential of this technique to be used in hematology and cyto-pathology studies aimed at identifying nanometer-sized features that cannot be distinguished otherwise with conventional optical microscopy.

  3. Fluorescent dyes with large Stokes shifts for super-resolution optical microscopy of biological objects: a review

    International Nuclear Information System (INIS)

    Sednev, Maksim V; Belov, Vladimir N; Hell, Stefan W

    2015-01-01

    The review deals with commercially available organic dyes possessing large Stokes shifts and their applications as fluorescent labels in optical microscopy based on stimulated emission depletion (STED). STED microscopy breaks Abbe’s diffraction barrier and provides optical resolution beyond the diffraction limit. STED microscopy is non-invasive and requires photostable fluorescent markers attached to biomolecules or other objects of interest. Up to now, in most biology-related STED experiments, bright and photoresistant dyes with small Stokes shifts of 20–40 nm were used. The rapid progress in STED microscopy showed that organic fluorophores possessing large Stokes shifts are indispensable in multi-color super-resolution techniques. The ultimate result of the imaging relies on the optimal combination of a dye, the bio-conjugation procedure and the performance of the optical microscope. Modern bioconjugation methods, basics of STED microscopy, as well as structures and spectral properties of the presently available fluorescent markers are reviewed and discussed. In particular, the spectral properties of the commercial dyes are tabulated and correlated with the available depletion wavelengths found in STED microscopes produced by LEICA Microsytems, Abberior Instruments and Picoquant GmbH. (topical review)

  4. Ultrafast ultrasound localization microscopy for deep super-resolution vascular imaging

    Science.gov (United States)

    Errico, Claudia; Pierre, Juliette; Pezet, Sophie; Desailly, Yann; Lenkei, Zsolt; Couture, Olivier; Tanter, Mickael

    2015-11-01

    Non-invasive imaging deep into organs at microscopic scales remains an open quest in biomedical imaging. Although optical microscopy is still limited to surface imaging owing to optical wave diffusion and fast decorrelation in tissue, revolutionary approaches such as fluorescence photo-activated localization microscopy led to a striking increase in resolution by more than an order of magnitude in the last decade. In contrast with optics, ultrasonic waves propagate deep into organs without losing their coherence and are much less affected by in vivo decorrelation processes. However, their resolution is impeded by the fundamental limits of diffraction, which impose a long-standing trade-off between resolution and penetration. This limits clinical and preclinical ultrasound imaging to a sub-millimetre scale. Here we demonstrate in vivo that ultrasound imaging at ultrafast frame rates (more than 500 frames per second) provides an analogue to optical localization microscopy by capturing the transient signal decorrelation of contrast agents—inert gas microbubbles. Ultrafast ultrasound localization microscopy allowed both non-invasive sub-wavelength structural imaging and haemodynamic quantification of rodent cerebral microvessels (less than ten micrometres in diameter) more than ten millimetres below the tissue surface, leading to transcranial whole-brain imaging within short acquisition times (tens of seconds). After intravenous injection, single echoes from individual microbubbles were detected through ultrafast imaging. Their localization, not limited by diffraction, was accumulated over 75,000 images, yielding 1,000,000 events per coronal plane and statistically independent pixels of ten micrometres in size. Precise temporal tracking of microbubble positions allowed us to extract accurately in-plane velocities of the blood flow with a large dynamic range (from one millimetre per second to several centimetres per second). These results pave the way for deep non

  5. Radial super-resolution in digital holographic microscopy using structured illumination with circular symmetry

    Science.gov (United States)

    Yin, Yujian; Su, Ping; Ma, Jianshe

    2018-01-01

    A method to improve the radial resolution using special structured light is proposed in the field of digital holographic microscopy (DHM). A specimen is illuminated with circular symmetrical structured light that makes the spectrum have radial movement, so that high frequency components of the specimen are moved into the passband of the receiver to overcome the diffraction limit. In the DHM imaging system, Computer Generated Hologram (CGH) technology is used to generate the required structured light grating. Then the grating is loaded into a spatial light modulator (SLM) to obtain specific structured illumination. After recording the hologram, digital reconstruction, for the microstructure of a binary optical element that needs to observe radial distribution, the radial resolution of the specimen is improved experimentally compare it with the result of one-dimensional sinusoidal structured light imaging. And a method of designing structured light is presented.

  6. Memory-effect based deconvolution microscopy for super-resolution imaging through scattering media

    Science.gov (United States)

    Edrei, Eitan; Scarcelli, Giuliano

    2016-09-01

    High-resolution imaging through turbid media is a fundamental challenge of optical sciences that has attracted a lot of attention in recent years for its wide range of potential applications. Here, we demonstrate that the resolution of imaging systems looking behind a highly scattering medium can be improved below the diffraction-limit. To achieve this, we demonstrate a novel microscopy technique enabled by the optical memory effect that uses a deconvolution image processing and thus it does not require iterative focusing, scanning or phase retrieval procedures. We show that this newly established ability of direct imaging through turbid media provides fundamental and practical advantages such as three-dimensional refocusing and unambiguous object reconstruction.

  7. Super-resolution

    DEFF Research Database (Denmark)

    Nasrollahi, Kamal; Moeslund, Thomas B.

    2014-01-01

    Super-resolution, the process of obtaining one or more high-resolution images from one or more low-resolution observations, has been a very attractive research topic over the last two decades. It has found practical applications in many real world problems in different fields, from satellite...

  8. Developing a New Biophysical Tool to Combine Magneto-Optical Tweezers with Super-Resolution Fluorescence Microscopy

    Directory of Open Access Journals (Sweden)

    Zhaokun Zhou

    2015-06-01

    Full Text Available We present a novel experimental setup in which magnetic and optical tweezers are combined for torque and force transduction onto single filamentous molecules in a transverse configuration to allow simultaneous mechanical measurement and manipulation. Previously we have developed a super-resolution imaging module which, in conjunction with advanced imaging techniques such as Blinking assisted Localisation Microscopy (BaLM, achieves localisation precision of single fluorescent dye molecules bound to DNA of ~30 nm along the contour of the molecule; our work here describes developments in producing a system which combines tweezing and super-resolution fluorescence imaging. The instrument also features an acousto-optic deflector that temporally divides the laser beam to form multiple traps for high throughput statistics collection. Our motivation for developing the new tool is to enable direct observation of detailed molecular topological transformation and protein binding event localisation in a stretching/twisting mechanical assay that previously could hitherto only be deduced indirectly from the end-to-end length variation of DNA. Our approach is simple and robust enough for reproduction in the lab without the requirement of precise hardware engineering, yet is capable of unveiling the elastic and dynamic properties of filamentous molecules that have been hidden using traditional tools.

  9. Antimicrobial agent triclosan disrupts mitochondrial structure, revealed by super-resolution microscopy, and inhibits mast cell signaling via calcium modulation.

    Science.gov (United States)

    Weatherly, Lisa M; Nelson, Andrew J; Shim, Juyoung; Riitano, Abigail M; Gerson, Erik D; Hart, Andrew J; de Juan-Sanz, Jaime; Ryan, Timothy A; Sher, Roger; Hess, Samuel T; Gosse, Julie A

    2018-06-15

    The antimicrobial agent triclosan (TCS) is used in products such as toothpaste and surgical soaps and is readily absorbed into oral mucosa and human skin. These and many other tissues contain mast cells, which are involved in numerous physiologies and diseases. Mast cells release chemical mediators through a process termed degranulation, which is inhibited by TCS. Investigation into the underlying mechanisms led to the finding that TCS is a mitochondrial uncoupler at non-cytotoxic, low-micromolar doses in several cell types and live zebrafish. Our aim was to determine the mechanisms underlying TCS disruption of mitochondrial function and of mast cell signaling. We combined super-resolution (fluorescence photoactivation localization) microscopy and multiple fluorescence-based assays to detail triclosan's effects in living mast cells, fibroblasts, and primary human keratinocytes. TCS disrupts mitochondrial nanostructure, causing mitochondria to undergo fission and to form a toroidal, "donut" shape. TCS increases reactive oxygen species production, decreases mitochondrial membrane potential, and disrupts ER and mitochondrial Ca 2+ levels, processes that cause mitochondrial fission. TCS is 60 × more potent than the banned uncoupler 2,4-dinitrophenol. TCS inhibits mast cell degranulation by decreasing mitochondrial membrane potential, disrupting microtubule polymerization, and inhibiting mitochondrial translocation, which reduces Ca 2+ influx into the cell. Our findings provide mechanisms for both triclosan's inhibition of mast cell signaling and its universal disruption of mitochondria. These mechanisms provide partial explanations for triclosan's adverse effects on human reproduction, immunology, and development. This study is the first to utilize super-resolution microscopy in the field of toxicology. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Spatio-temporal image correlation spectroscopy and super-resolution microscopy to quantify molecular dynamics in T cells.

    Science.gov (United States)

    Ashdown, George W; Owen, Dylan M

    2018-02-02

    Many cellular processes are regulated by the spatio-temporal organisation of signalling complexes, cytoskeletal components and membranes. One such example is at the T cell immunological synapse where the retrograde flow of cortical filamentous (F)-actin from the synapse periphery drives signalling protein microclusters towards the synapse centre. The density of this mesh however, makes visualisation and analysis of individual actin fibres difficult due to the resolution limit of conventional microscopy. Recently, super-resolution methods such as structured illumination microscopy (SIM) have surpassed this resolution limit. Here, we apply SIM to better visualise the dense cortical actin meshwork in T cell synapses formed against activating, antibody-coated surfaces and image under total-internal reflection fluorescence (TIRF) illumination. To analyse the observed molecular flows, and the relationship between them, we apply spatio-temporal image correlation spectroscopy (STICS) and its cross-correlation variant (STICCS). We show that the dynamic cortical actin mesh can be visualised with unprecedented detail and that STICS/STICCS can output accurate, quantitative maps of molecular flow velocity and directionality from such data. We find that the actin flow can be disrupted using small molecule inhibitors of actin polymerisation. This combination of imaging and quantitative analysis may provide an important new tool for researchers to investigate the molecular dynamics at cellular length scales. Here we demonstrate the retrograde flow of F-actin which may be important for the clustering and dynamics of key signalling proteins within the plasma membrane, a phenomenon which is vital to correct T cell activation and therefore the mounting of an effective immune response. Copyright © 2018. Published by Elsevier Inc.

  11. The super-resolution debate

    Science.gov (United States)

    Won, Rachel

    2018-05-01

    In the quest for nanoscopy with super-resolution, consensus from the imaging community is that super-resolution is not always needed and that scientists should choose an imaging technique based on their specific application.

  12. Tilted Light Sheet Microscopy with 3D Point Spread Functions for Single-Molecule Super-Resolution Imaging in Mammalian Cells.

    Science.gov (United States)

    Gustavsson, Anna-Karin; Petrov, Petar N; Lee, Maurice Y; Shechtman, Yoav; Moerner, W E

    2018-02-01

    To obtain a complete picture of subcellular nanostructures, cells must be imaged with high resolution in all three dimensions (3D). Here, we present tilted light sheet microscopy with 3D point spread functions (TILT3D), an imaging platform that combines a novel, tilted light sheet illumination strategy with engineered long axial range point spread functions (PSFs) for low-background, 3D super localization of single molecules as well as 3D super-resolution imaging in thick cells. TILT3D is built upon a standard inverted microscope and has minimal custom parts. The axial positions of the single molecules are encoded in the shape of the PSF rather than in the position or thickness of the light sheet, and the light sheet can therefore be formed using simple optics. The result is flexible and user-friendly 3D super-resolution imaging with tens of nm localization precision throughout thick mammalian cells. We validated TILT3D for 3D super-resolution imaging in mammalian cells by imaging mitochondria and the full nuclear lamina using the double-helix PSF for single-molecule detection and the recently developed Tetrapod PSF for fiducial bead tracking and live axial drift correction. We envision TILT3D to become an important tool not only for 3D super-resolution imaging, but also for live whole-cell single-particle and single-molecule tracking.

  13. Quantitating morphological changes in biological samples during scanning electron microscopy sample preparation with correlative super-resolution microscopy.

    Science.gov (United States)

    Zhang, Ying; Huang, Tao; Jorgens, Danielle M; Nickerson, Andrew; Lin, Li-Jung; Pelz, Joshua; Gray, Joe W; López, Claudia S; Nan, Xiaolin

    2017-01-01

    Sample preparation is critical to biological electron microscopy (EM), and there have been continuous efforts on optimizing the procedures to best preserve structures of interest in the sample. However, a quantitative characterization of the morphological changes associated with each step in EM sample preparation is currently lacking. Using correlative EM and superresolution microscopy (SRM), we have examined the effects of different drying methods as well as osmium tetroxide (OsO4) post-fixation on cell morphology during scanning electron microscopy (SEM) sample preparation. Here, SRM images of the sample acquired under hydrated conditions were used as a baseline for evaluating morphological changes as the sample went through SEM sample processing. We found that both chemical drying and critical point drying lead to a mild cellular boundary retraction of ~60 nm. Post-fixation by OsO4 causes at least 40 nm additional boundary retraction. We also found that coating coverslips with adhesion molecules such as fibronectin prior to cell plating helps reduce cell distortion from OsO4 post-fixation. These quantitative measurements offer useful information for identifying causes of cell distortions in SEM sample preparation and improving current procedures.

  14. Nanoscale clustering of the neurotrophin receptor TrkB revealed by super-resolution STED microscopy

    Czech Academy of Sciences Publication Activity Database

    Angelov, Borislav; Angelova, A.

    2017-01-01

    Roč. 9, č. 28 (2017), s. 9797-9804 ISSN 2040-3364 R&D Projects: GA MŠk EF15_003/0000447; GA MŠk EF15_008/0000162; GA ČR(CZ) GA17-00973S Grant - others:OP VVV - ELIBIO(XE) CZ.02.1.01/0.0/0.0/15_003/0000447; ELI Beamlines(XE) CZ.02.1.01/0.0/0.0/15_008/0000162 Institutional support: RVO:68378271 Keywords : emission depletion microscopy * higher-order oligomers * neurodegenerative diseases * mechanistic insights Subject RIV: BL - Plasma and Gas Discharge Physics OBOR OECD: Fluids and plasma physics (including surface physics) Impact factor: 7.367, year: 2016

  15. Imaging lipid domains in cell membranes: the advent of super-resolution fluorescence microscopy

    Directory of Open Access Journals (Sweden)

    Dylan Myers Owen

    2013-12-01

    Full Text Available The lipid bilayer of model membranes, liposomes reconstituted from cell lipids, and plasma membrane vesicles and spheres can separate into two distinct liquid phases to yield lipid domains with liquid-ordered and liquid-disordered properties. These observations are the basis of the lipid raft hypothesis that postulates the existence of cholesterol-enriched ordered-phase lipid domains in cell membranes that could regulate protein mobility, localization and interaction. Here we review the evidence that nano-scaled lipid complexes and meso-scaled lipid domains exist in cell membranes and how new fluorescence microscopy techniques that overcome the diffraction limit provide new insights into lipid organization in cell membranes.

  16. Super-resolution microscopy reveals presynaptic localization of the ALS / FTD related protein FUS in hippocampal neurons

    Directory of Open Access Journals (Sweden)

    Michael eSchoen

    2016-01-01

    Full Text Available Fused in Sarcoma (FUS is a multifunctional RNA- / DNA-binding protein, which is involved in the pathogenesis of the neurodegenerative disorders amyotrophic lateral sclerosis (ALS and frontotemporal dementia (FTD. A common hallmark of these disorders is the abnormal accumulation of mutated FUS protein in the cytoplasm. Under normal conditions FUS is confined to the nuclear compartment, in neurons however, additional somatodendritic localization can be observed. In this study, we carefully analyzed the subcellular localization of endogenous FUS at synaptic sites of hippocampal neurons which are among the most affected cell types in frontotemporal dementia with FUS pathology. We could confirm a strong nuclear localization of FUS as well as its prominent and widespread neuronal expression throughout the adult and developing rat brain, particularly in the hippocampus, the cerebellum and the outer layers of the cortex. Intriguingly, FUS was also consistently observed at synaptic sites as detected by neuronal subcellular fractionation as well as by immunolabeling. To define a pre- and / or postsynaptic localization of FUS, we employed super-resolution fluorescence localization microscopy. FUS was found to be localized within the axon terminal in close proximity to the presynaptic vesicle protein Synaptophysin1 and adjacent to the active zone protein Bassoon, but well separated from the postsynaptic protein PSD-95. Having shown the presynaptic localization of FUS in the nervous system, a novel extranuclear role of FUS at neuronal contact sites has to be considered. Since there is growing evidence that local presynaptic translation might also be an important mechanism for plasticity, FUS - like the fragile X mental retardation protein FMRP - might act as one of the presynaptic RNA-binding proteins regulating this machinery. Our observation of presynaptic FUS should foster further investigations to determine its role in neurodegenerative diseases such as

  17. Tilted light sheet microscopy with 3D point spread functions for single-molecule super-resolution imaging in mammalian cells

    Science.gov (United States)

    Gustavsson, Anna-Karin; Petrov, Petar N.; Lee, Maurice Y.; Shechtman, Yoav; Moerner, W. E.

    2018-02-01

    To obtain a complete picture of subcellular nanostructures, cells must be imaged with high resolution in all three dimensions (3D). Here, we present tilted light sheet microscopy with 3D point spread functions (TILT3D), an imaging platform that combines a novel, tilted light sheet illumination strategy with engineered long axial range point spread functions (PSFs) for low-background, 3D super localization of single molecules as well as 3D super-resolution imaging in thick cells. TILT3D is built upon a standard inverted microscope and has minimal custom parts. The axial positions of the single molecules are encoded in the shape of the PSF rather than in the position or thickness of the light sheet, and the light sheet can therefore be formed using simple optics. The result is flexible and user-friendly 3D super-resolution imaging with tens of nm localization precision throughout thick mammalian cells. We validated TILT3D for 3D superresolution imaging in mammalian cells by imaging mitochondria and the full nuclear lamina using the double-helix PSF for single-molecule detection and the recently developed Tetrapod PSF for fiducial bead tracking and live axial drift correction. We envision TILT3D to become an important tool not only for 3D super-resolution imaging, but also for live whole-cell single-particle and single-molecule tracking.

  18. Quantitative super-resolution localization microscopy of DNA in situ using Vybrant® DyeCycle™ Violet fluorescent probe

    Directory of Open Access Journals (Sweden)

    Dominika Żurek-Biesiada

    2016-06-01

    Full Text Available Single Molecule Localization Microscopy (SMLM is a recently emerged optical imaging method that was shown to achieve a resolution in the order of tens of nanometers in intact cells. Novel high resolution imaging methods might be crucial for understanding of how the chromatin, a complex of DNA and proteins, is arranged in the eukaryotic cell nucleus. Such an approach utilizing switching of a fluorescent, DNA-binding dye Vybrant® DyeCycle™ Violet has been previously demonstrated by us (Żurek-Biesiada et al., 2015 [1]. Here we provide quantitative information on the influence of the chemical environment on the behavior of the dye, discuss the variability in the DNA-associated signal density, and demonstrate direct proof of enhanced structural resolution. Furthermore, we compare different visualization approaches. Finally, we describe various opportunities of multicolor DNA/SMLM imaging in eukaryotic cell nuclei.

  19. Cortical actin nodes: Their dynamics and recruitment of podosomal proteins as revealed by super-resolution and single-molecule microscopy

    Science.gov (United States)

    Shirai, Yuki M.; Tsunoyama, Taka A.; Hiramoto-Yamaki, Nao; Hirosawa, Koichiro M.; Shibata, Akihiro C. E.; Kondo, Kenichi; Tsurumune, Atsushi; Ishidate, Fumiyoshi; Kusumi, Akihiro

    2017-01-01

    Electron tomography of the plasma membrane (PM) identified several layers of cortical actin meshwork running parallel to the PM cytoplasmic surface throughout the PM. Here, cortical actin structures and dynamics were examined in living cells, using super-resolution microscopy, with (x,y)- and z-resolutions of ~140 and ~400 nm, respectively, and single-molecule imaging. The super-resolution microscopy identified sub-micron-sized actin clusters that appeared identical by both phalloidin post-fixation staining and Lifeact-mGFP expression followed by fixation, and therefore, these actin clusters were named “actin-pl-clusters”. In live cells, the actin-pl-clusters visualized by Lifeact-mGFP linked two or more actin filaments in the fine actin meshwork, acting as a node of the meshwork, and dynamically moved on/along the meshwork in a myosin II-dependent manner. Their formation depended on the Arp2/3 activities, suggesting that the movements could involve both the myosin motor activity and actin polymerization-depolymerization. The actin-pl-clusters differ from the actin nodes/asters found previously after latrunculin treatments, since myosin II and filamin A were not colocalized with the actin-pl-clusters, and the actin-pl-clusters were much smaller than the previously reported nodes/asters. The Lifeact linked to a fluorescently-labeled transmembrane peptide from syntaxin4 (Lifeact-TM) expressed in the PM exhibited temporary immobilization in the PM regions on which actin-pl-clusters and stress fibers were projected, showing that ≥66% of actin-pl-clusters and 89% of stress fibers were located in close proximity (within 3.5 nm) to the PM cytoplasmic surface. Podosome-associated cytoplasmic proteins, Tks4, Tks5, cortactin, and N-WASP, were transiently recruited to actin-pl-clusters, and thus, we propose that actin-pl-clusters also represent “actin podosome-like clusters”. PMID:29190677

  20. The journey of integrins and partners in a complex interactions landscape studied by super-resolution microscopy and single protein tracking

    International Nuclear Information System (INIS)

    Rossier, Olivier; Giannone, Grégory

    2016-01-01

    Cells adjust their adhesive and cytoskeletal organizations according to changes in the biochemical and physical nature of their surroundings. In return, by adhering and generating forces on the extracellular matrix (ECM) cells organize their microenvironment. Integrin-dependent focal adhesions (FAs) are the converging zones integrating biochemical and biomechanical signals arising from the ECM and the actin cytoskeleton. Thus, integrin-mediated adhesion and mechanotransduction, the conversion of mechanical forces into biochemical signals, are involved in critical cellular functions such as migration, proliferation and differentiation, and their deregulation contributes to pathologies including cancer. A challenging problem is to decipher how stochastic protein movements and interactions lead to formation of dynamic architecture such as integrin-dependent adhesive structures. In this review, we will describe recent advances made possible by super-resolution microscopies and single molecule tracking approaches that provided new understanding on the organization and the dynamics of integrins and intracellular regulators at the nanoscale in living cells.

  1. The journey of integrins and partners in a complex interactions landscape studied by super-resolution microscopy and single protein tracking

    Energy Technology Data Exchange (ETDEWEB)

    Rossier, Olivier; Giannone, Grégory [Univ. Bordeaux, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux (France); CNRS, Interdisciplinary Institute for Neuroscience, UMR 5297, F-33000 Bordeaux (France)

    2016-04-10

    Cells adjust their adhesive and cytoskeletal organizations according to changes in the biochemical and physical nature of their surroundings. In return, by adhering and generating forces on the extracellular matrix (ECM) cells organize their microenvironment. Integrin-dependent focal adhesions (FAs) are the converging zones integrating biochemical and biomechanical signals arising from the ECM and the actin cytoskeleton. Thus, integrin-mediated adhesion and mechanotransduction, the conversion of mechanical forces into biochemical signals, are involved in critical cellular functions such as migration, proliferation and differentiation, and their deregulation contributes to pathologies including cancer. A challenging problem is to decipher how stochastic protein movements and interactions lead to formation of dynamic architecture such as integrin-dependent adhesive structures. In this review, we will describe recent advances made possible by super-resolution microscopies and single molecule tracking approaches that provided new understanding on the organization and the dynamics of integrins and intracellular regulators at the nanoscale in living cells.

  2. High-resolution electron microscopy

    CERN Document Server

    Spence, John C H

    2013-01-01

    This new fourth edition of the standard text on atomic-resolution transmission electron microscopy (TEM) retains previous material on the fundamentals of electron optics and aberration correction, linear imaging theory (including wave aberrations to fifth order) with partial coherence, and multiple-scattering theory. Also preserved are updated earlier sections on practical methods, with detailed step-by-step accounts of the procedures needed to obtain the highest quality images of atoms and molecules using a modern TEM or STEM electron microscope. Applications sections have been updated - these include the semiconductor industry, superconductor research, solid state chemistry and nanoscience, and metallurgy, mineralogy, condensed matter physics, materials science and material on cryo-electron microscopy for structural biology. New or expanded sections have been added on electron holography, aberration correction, field-emission guns, imaging filters, super-resolution methods, Ptychography, Ronchigrams, tomogr...

  3. Super-resolution microscopy reveals functional organization of dopamine transporters into cholesterol and neuronal activity-dependent nanodomains

    DEFF Research Database (Denmark)

    Rahbek-Clemmensen, Troels; Lycas, Matthew D.; Erlendsson, Simon

    2017-01-01

    is dynamically sequestrated into cholesterol-dependent nanodomains in the plasma membrane of presynaptic varicosities and neuronal projections of dopaminergic neurons. Stochastic optical reconstruction microscopy reveals irregular dopamine transporter nanodomains (∼70 nm mean diameter) that were highly sensitive...... to cholesterol depletion. Live photoactivated localization microscopy shows a similar dopamine transporter membrane organization in live heterologous cells. In neurons, dual-color dSTORM shows that tyrosine hydroxylase and vesicular monoamine transporter-2 are distinctively localized adjacent to...

  4. Super-resolved terahertz microscopy by knife-edge scan

    Science.gov (United States)

    Giliberti, V.; Flammini, M.; Ciano, C.; Pontecorvo, E.; Del Re, E.; Ortolani, M.

    2017-08-01

    We present a compact, all solid-state THz confocal microscope operating at 0.30 THz that achieves super-resolution by using the knife-edge scan approach. In the final reconstructed image, a lateral resolution of 60 μm ≍ λ/17 is demonstrated when the knife-edge is deep in the near-field of the sample surface. When the knife-edge is lifted up to λ/4 from the sample surface, a certain degree of super-resolution is maintained with a resolution of 0.4 mm, i.e. more than a factor 2 if compared to the diffraction-limited scheme. The present results open an interesting path towards super-resolved imaging with in-depth information that would be peculiar to THz microscopy systems.

  5. The journey of integrins and partners in a complex interactions landscape studied by super-resolution microscopy and single protein tracking.

    Science.gov (United States)

    Rossier, Olivier; Giannone, Grégory

    2016-04-10

    Cells adjust their adhesive and cytoskeletal organizations according to changes in the biochemical and physical nature of their surroundings. In return, by adhering and generating forces on the extracellular matrix (ECM) cells organize their microenvironment. Integrin-dependent focal adhesions (FAs) are the converging zones integrating biochemical and biomechanical signals arising from the ECM and the actin cytoskeleton. Thus, integrin-mediated adhesion and mechanotransduction, the conversion of mechanical forces into biochemical signals, are involved in critical cellular functions such as migration, proliferation and differentiation, and their deregulation contributes to pathologies including cancer. A challenging problem is to decipher how stochastic protein movements and interactions lead to formation of dynamic architecture such as integrin-dependent adhesive structures. In this review, we will describe recent advances made possible by super-resolution microscopies and single molecule tracking approaches that provided new understanding on the organization and the dynamics of integrins and intracellular regulators at the nanoscale in living cells. Copyright © 2015. Published by Elsevier Inc.

  6. Localization-based super-resolution imaging of cellular structures.

    Science.gov (United States)

    Kanchanawong, Pakorn; Waterman, Clare M

    2013-01-01

    Fluorescence microscopy allows direct visualization of fluorescently tagged proteins within cells. However, the spatial resolution of conventional fluorescence microscopes is limited by diffraction to ~250 nm, prompting the development of super-resolution microscopy which offers resolution approaching the scale of single proteins, i.e., ~20 nm. Here, we describe protocols for single molecule localization-based super-resolution imaging, using focal adhesion proteins as an example and employing either photoswitchable fluorophores or photoactivatable fluorescent proteins. These protocols should also be easily adaptable to imaging a broad array of macromolecular assemblies in cells whose components can be fluorescently tagged and assemble into high density structures.

  7. Super-resolution microscopy reveals the insulin-resistance-regulated reorganization of GLUT4 on plasma membranes.

    Science.gov (United States)

    Gao, Lan; Chen, Junling; Gao, Jing; Wang, Hongda; Xiong, Wenyong

    2017-01-15

    GLUT4 (also known as SLC2A4) is essential for glucose uptake in skeletal muscles and adipocytes, which play central roles in whole-body glucose metabolism. Here, using direct stochastic optical reconstruction microscopy (dSTORM) to investigate the characteristics of plasma-membrane-fused GLUT4 at the single-molecule level, we have demonstrated that insulin and insulin resistance regulate the spatial organization of GLUT4 in adipocytes. Stimulation with insulin shifted the balance of GLUT4 on the plasma membrane toward a more dispersed configuration. In contrast, insulin resistance induced a more clustered distribution of GLUT4 and increased the mean number of molecules per cluster. Furthermore, our data demonstrate that the F 5 QQI motif and lipid rafts mediate the maintenance of GLUT4 clusters on the plasma membrane. Mutation of F 5 QQI (F 5 QQA-GLUT4) induced a more clustered distribution of GLUT4; moreover, destruction of lipid rafts in adipocytes expressing F 5 QQA-GLUT4 dramatically decreased the percentage of large clusters and the mean number of molecules per cluster. In conclusion, our data clarify the effects of insulin stimulation or insulin resistance on GLUT4 reorganization on the plasma membrane and reveal new pathogenic mechanisms of insulin resistance. © 2017. Published by The Company of Biologists Ltd.

  8. Perspectives in Super-resolved Fluorescence Microscopy: What comes next?

    Science.gov (United States)

    Cremer, Christoph; Birk, Udo

    2016-04-01

    The Nobel Prize in Chemistry 2014 has been awarded to three scientists involved in the development of STED and PALM super-resolution fluorescence microscopy (SRM) methods. They have proven that it is possible to overcome the hundred year old theoretical limit for the resolution potential of light microscopy (of about 200 nm for visible light), which for decades has precluded a direct glimpse of the molecular machinery of life. None of the present-day super-resolution techniques have invalidated the Abbe limit for light optical detection; however, they have found clever ways around it. In this report, we discuss some of the challenges still to be resolved before arising SRM approaches will be fit to bring about the revolution in Biology and Medicine envisaged. Some of the challenges discussed are the applicability to image live and/or large samples, the further enhancement of resolution, future developments of labels, and multi-spectral approaches.

  9. Perspectives in Super-resolved Fluorescence Microscopy: What comes next?

    Directory of Open Access Journals (Sweden)

    Christoph eCremer

    2016-04-01

    Full Text Available The Nobel Prize in Chemistry 2014 has been awarded to three scientists involved in the development of STED and PALM super-resolution fluorescence microscopy (SRM methods. They have proven that it is possible to overcome the hundred year old theoretical limit for the resolution potential of light microscopy (of about 200 nm for visible light, which for decades has precluded a direct glimpse of the molecular machinery of life. None of the present-day super-resolution techniques have invalidated the Abbe limit for light optical detection; however, they have found clever ways around it. In this report, we discuss some of the challenges still to be resolved before arising SRM approaches will be fit to bring about the revolution in Biology and Medicine envisaged. Some of the challenges discussed are the applicability to image live and/or large samples, the further enhancement of resolution, future developments of labels, and multi-spectral approaches.

  10. Subnuclear localization, rates and effectiveness of UVC-induced unscheduled DNA synthesis visualized by fluorescence widefield, confocal and super-resolution microscopy.

    Science.gov (United States)

    Pierzyńska-Mach, Agnieszka; Szczurek, Aleksander; Cella Zanacchi, Francesca; Pennacchietti, Francesca; Drukała, Justyna; Diaspro, Alberto; Cremer, Christoph; Darzynkiewicz, Zbigniew; Dobrucki, Jurek W

    2016-01-01

    Unscheduled DNA synthesis (UDS) is the final stage of the process of repair of DNA lesions induced by UVC. We detected UDS using a DNA precursor, 5-ethynyl-2'-deoxyuridine (EdU). Using wide-field, confocal and super-resolution fluorescence microscopy and normal human fibroblasts, derived from healthy subjects, we demonstrate that the sub-nuclear pattern of UDS detected via incorporation of EdU is different from that when BrdU is used as DNA precursor. EdU incorporation occurs evenly throughout chromatin, as opposed to just a few small and large repair foci detected by BrdU. We attribute this difference to the fact that BrdU antibody is of much larger size than EdU, and its accessibility to the incorporated precursor requires the presence of denatured sections of DNA. It appears that under the standard conditions of immunocytochemical detection of BrdU only fragments of DNA of various length are being denatured. We argue that, compared with BrdU, the UDS pattern visualized by EdU constitutes a more faithful representation of sub-nuclear distribution of the final stage of nucleotide excision repair induced by UVC. Using the optimized integrated EdU detection procedure we also measured the relative amount of the DNA precursor incorporated by cells during UDS following exposure to various doses of UVC. Also described is the high degree of heterogeneity in terms of the UVC-induced EdU incorporation per cell, presumably reflecting various DNA repair efficiencies or differences in the level of endogenous dT competing with EdU within a population of normal human fibroblasts.

  11. Hypotonic activation of the myo-inositol transporter SLC5A3 in HEK293 cells probed by cell volumetry, confocal and super-resolution microscopy.

    Directory of Open Access Journals (Sweden)

    Joseph Andronic

    Full Text Available Swelling-activated pathways for myo-inositol, one of the most abundant organic osmolytes in mammalian cells, have not yet been identified. The present study explores the SLC5A3 protein as a possible transporter of myo-inositol in hyponically swollen HEK293 cells. To address this issue, we examined the relationship between the hypotonicity-induced changes in plasma membrane permeability to myo-inositol P ino [m/s] and expression/localization of SLC5A3. P ino values were determined by cell volumetry over a wide tonicity range (100-275 mOsm in myo-inositol-substituted solutions. While being negligible under mild hypotonicity (200-275 mOsm, P ino grew rapidly at osmolalities below 200 mOsm to reach a maximum of ∼ 3 nm/s at 100-125 mOsm, as indicated by fast cell swelling due to myo-inositol influx. The increase in P ino resulted most likely from the hypotonicity-mediated incorporation of cytosolic SLC5A3 into the plasma membrane, as revealed by confocal fluorescence microscopy of cells expressing EGFP-tagged SLC5A3 and super-resolution imaging of immunostained SLC5A3 by direct stochastic optical reconstruction microscopy (dSTORM. dSTORM in hypotonic cells revealed a surface density of membrane-associated SLC5A3 proteins of 200-2000 localizations/μm2. Assuming SLC5A3 to be the major path for myo-inositol, a turnover rate of 80-800 myo-inositol molecules per second for a single transporter protein was estimated from combined volumetric and dSTORM data. Hypotonic stress also caused a significant upregulation of SLC5A3 gene expression as detected by semiquantitative RT-PCR and Western blot analysis. In summary, our data provide first evidence for swelling-mediated activation of SLC5A3 thus suggesting a functional role of this transporter in hypotonic volume regulation of mammalian cells.

  12. Super resolution for astronomical observations

    Science.gov (United States)

    Li, Zhan; Peng, Qingyu; Bhanu, Bir; Zhang, Qingfeng; He, Haifeng

    2018-05-01

    In order to obtain detailed information from multiple telescope observations a general blind super-resolution (SR) reconstruction approach for astronomical images is proposed in this paper. A pixel-reliability-based SR reconstruction algorithm is described and implemented, where the developed process incorporates flat field correction, automatic star searching and centering, iterative star matching, and sub-pixel image registration. Images captured by the 1-m telescope at Yunnan Observatory are used to test the proposed technique. The results of these experiments indicate that, following SR reconstruction, faint stars are more distinct, bright stars have sharper profiles, and the backgrounds have higher details; thus these results benefit from the high-precision star centering and image registration provided by the developed method. Application of the proposed approach not only provides more opportunities for new discoveries from astronomical image sequences, but will also contribute to enhancing the capabilities of most spatial or ground-based telescopes.

  13. A novel super-resolution camera model

    Science.gov (United States)

    Shao, Xiaopeng; Wang, Yi; Xu, Jie; Wang, Lin; Liu, Fei; Luo, Qiuhua; Chen, Xiaodong; Bi, Xiangli

    2015-05-01

    Aiming to realize super resolution(SR) to single image and video reconstruction, a super resolution camera model is proposed for the problem that the resolution of the images obtained by traditional cameras behave comparatively low. To achieve this function we put a certain driving device such as piezoelectric ceramics in the camera. By controlling the driving device, a set of continuous low resolution(LR) images can be obtained and stored instantaneity, which reflect the randomness of the displacements and the real-time performance of the storage very well. The low resolution image sequences have different redundant information and some particular priori information, thus it is possible to restore super resolution image factually and effectively. The sample method is used to derive the reconstruction principle of super resolution, which analyzes the possible improvement degree of the resolution in theory. The super resolution algorithm based on learning is used to reconstruct single image and the variational Bayesian algorithm is simulated to reconstruct the low resolution images with random displacements, which models the unknown high resolution image, motion parameters and unknown model parameters in one hierarchical Bayesian framework. Utilizing sub-pixel registration method, a super resolution image of the scene can be reconstructed. The results of 16 images reconstruction show that this camera model can increase the image resolution to 2 times, obtaining images with higher resolution in currently available hardware levels.

  14. Single Image Super Resolution via Sparse Reconstruction

    NARCIS (Netherlands)

    Kruithof, M.C.; Eekeren, A.W.M. van; Dijk, J.; Schutte, K.

    2012-01-01

    High resolution sensors are required for recognition purposes. Low resolution sensors, however, are still widely used. Software can be used to increase the resolution of such sensors. One way of increasing the resolution of the images produced is using multi-frame super resolution algorithms.

  15. Three-dimensional super-resolution microscopy of the inactive X chromosome territory reveals a collapse of its active nuclear compartment harboring distinct Xist RNA foci.

    Science.gov (United States)

    Smeets, Daniel; Markaki, Yolanda; Schmid, Volker J; Kraus, Felix; Tattermusch, Anna; Cerase, Andrea; Sterr, Michael; Fiedler, Susanne; Demmerle, Justin; Popken, Jens; Leonhardt, Heinrich; Brockdorff, Neil; Cremer, Thomas; Schermelleh, Lothar; Cremer, Marion

    2014-01-01

    A Xist RNA decorated Barr body is the structural hallmark of the compacted inactive X territory in female mammals. Using super-resolution three-dimensional structured illumination microscopy (3D-SIM) and quantitative image analysis, we compared its ultrastructure with active chromosome territories (CTs) in human and mouse somatic cells, and explored the spatio-temporal process of Barr body formation at onset of inactivation in early differentiating mouse embryonic stem cells (ESCs). We demonstrate that all CTs are composed of structurally linked chromatin domain clusters (CDCs). In active CTs the periphery of CDCs harbors low-density chromatin enriched with transcriptionally competent markers, called the perichromatin region (PR). The PR borders on a contiguous channel system, the interchromatin compartment (IC), which starts at nuclear pores and pervades CTs. We propose that the PR and macromolecular complexes in IC channels together form the transcriptionally permissive active nuclear compartment (ANC). The Barr body differs from active CTs by a partially collapsed ANC with CDCs coming significantly closer together, although a rudimentary IC channel system connected to nuclear pores is maintained. Distinct Xist RNA foci, closely adjacent to the nuclear matrix scaffold attachment factor-A (SAF-A) localize throughout Xi along the rudimentary ANC. In early differentiating ESCs initial Xist RNA spreading precedes Barr body formation, which occurs concurrent with the subsequent exclusion of RNA polymerase II (RNAP II). Induction of a transgenic autosomal Xist RNA in a male ESC triggers the formation of an 'autosomal Barr body' with less compacted chromatin and incomplete RNAP II exclusion. 3D-SIM provides experimental evidence for profound differences between the functional architecture of transcriptionally active CTs and the Barr body. Basic structural features of CT organization such as CDCs and IC channels are however still recognized, arguing against a uniform

  16. Microsphere-based super-resolution scanning optical microscope.

    Science.gov (United States)

    Huszka, Gergely; Yang, Hui; Gijs, Martin A M

    2017-06-26

    High-refractive index dielectric microspheres positioned within the field of view of a microscope objective in a dielectric medium can focus the light into a so-called photonic nanojet. A sample placed in such nanojet can be imaged by the objective with super-resolution, i.e. with a resolution beyond the classical diffraction limit. However, when imaging nanostructures on a substrate, the propagation distance of a light wave in the dielectric medium in between the substrate and the microsphere must be small enough to reveal the sample's nanometric features. Therefore, only the central part of an image obtained through a microsphere shows super-resolution details, which are typically ∼100 nm using white light (peak at λ = 600 nm). We have performed finite element simulations of the role of this critical distance in the super-resolution effect. Super-resolution imaging of a sample placed beneath the microsphere is only possible within a very restricted central area of ∼10 μm 2 , where the separation distance between the substrate and the microsphere surface is very small (∼1 μm). To generate super-resolution images over larger areas of the sample, we have fixed a microsphere on a frame attached to the microscope objective, which is automatically scanned over the sample in a step-by-step fashion. This generates a set of image tiles, which are subsequently stitched into a single super-resolution image (with resolution of λ/4-λ/5) of a sample area of up to ∼10 4 μm 2 . Scanning a standard optical microscope objective with microsphere therefore enables super-resolution microscopy over the complete field-of-view of the objective.

  17. Focusing super resolution on the cytoskeleton [version 1; referees: 3 approved

    Directory of Open Access Journals (Sweden)

    Eric A. Shelden

    2016-05-01

    Full Text Available Super resolution imaging is becoming an increasingly important tool in the arsenal of methods available to cell biologists. In recognition of its potential, the Nobel Prize for chemistry was awarded to three investigators involved in the development of super resolution imaging methods in 2014. The availability of commercial instruments for super resolution imaging has further spurred the development of new methods and reagents designed to take advantage of super resolution techniques. Super resolution offers the advantages traditionally associated with light microscopy, including the use of gentle fixation and specimen preparation methods, the ability to visualize multiple elements within a single specimen, and the potential to visualize dynamic changes in living specimens over time. However, imaging of living cells over time is difficult and super resolution imaging is computationally demanding. In this review, we discuss the advantages/disadvantages of different super resolution systems for imaging fixed live specimens, with particular regard to cytoskeleton structures.

  18. Learning from errors in super-resolution.

    Science.gov (United States)

    Tang, Yi; Yuan, Yuan

    2014-11-01

    A novel framework of learning-based super-resolution is proposed by employing the process of learning from the estimation errors. The estimation errors generated by different learning-based super-resolution algorithms are statistically shown to be sparse and uncertain. The sparsity of the estimation errors means most of estimation errors are small enough. The uncertainty of the estimation errors means the location of the pixel with larger estimation error is random. Noticing the prior information about the estimation errors, a nonlinear boosting process of learning from these estimation errors is introduced into the general framework of the learning-based super-resolution. Within the novel framework of super-resolution, a low-rank decomposition technique is used to share the information of different super-resolution estimations and to remove the sparse estimation errors from different learning algorithms or training samples. The experimental results show the effectiveness and the efficiency of the proposed framework in enhancing the performance of different learning-based algorithms.

  19. Re-scan confocal microscopy: scanning twice for better resolution.

    Science.gov (United States)

    De Luca, Giulia M R; Breedijk, Ronald M P; Brandt, Rick A J; Zeelenberg, Christiaan H C; de Jong, Babette E; Timmermans, Wendy; Azar, Leila Nahidi; Hoebe, Ron A; Stallinga, Sjoerd; Manders, Erik M M

    2013-01-01

    We present a new super-resolution technique, Re-scan Confocal Microscopy (RCM), based on standard confocal microscopy extended with an optical (re-scanning) unit that projects the image directly on a CCD-camera. This new microscope has improved lateral resolution and strongly improved sensitivity while maintaining the sectioning capability of a standard confocal microscope. This simple technology is typically useful for biological applications where the combination high-resolution and high-sensitivity is required.

  20. Temporal super resolution using variational methods

    DEFF Research Database (Denmark)

    Keller, Sune Høgild; Lauze, Francois Bernard; Nielsen, Mads

    2010-01-01

    Temporal super resolution (TSR) is the ability to convert video from one frame rate to another and is as such a key functionality in modern video processing systems. A higher frame rate than what is recorded is desired for high frame rate displays, for super slow-motion, and for video/film format...... observed when watching video on large and bright displays where the motion of high contrast edges often seem jerky and unnatural. A novel motion compensated (MC) TSR algorithm using variational methods for both optical flow calculation and the actual new frame interpolation is presented. The flow...

  1. Re-scan confocal microscopy (RCM) improves the resolution of confocal microscopy and increases the sensitivity

    NARCIS (Netherlands)

    de Luca, Giulia; Breedijk, Ronald; Hoebe, Ron; Stallinga, Sjoerd; Manders, Erik

    2017-01-01

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial

  2. Re-scan confocal microscopy (RCM) improves the resolution of confocal microscopy and increases the sensitivity

    NARCIS (Netherlands)

    De Luca, G.; Breedijk, R.; Hoebe, R.; Stallinga, S.; Manders, E.

    Re-scan confocal microscopy (RCM) is a new super-resolution technique based on a standard confocal microscope extended with a re-scan unit in the detection path that projects the emitted light onto a sensitive camera. In this paper the fundamental properties of RCM, lateral resolution, axial

  3. Super Resolution Algorithm for CCTVs

    Science.gov (United States)

    Gohshi, Seiichi

    2015-03-01

    Recently, security cameras and CCTV systems have become an important part of our daily lives. The rising demand for such systems has created business opportunities in this field, especially in big cities. Analogue CCTV systems are being replaced by digital systems, and HDTV CCTV has become quite common. HDTV CCTV can achieve images with high contrast and decent quality if they are clicked in daylight. However, the quality of an image clicked at night does not always have sufficient contrast and resolution because of poor lighting conditions. CCTV systems depend on infrared light at night to compensate for insufficient lighting conditions, thereby producing monochrome images and videos. However, these images and videos do not have high contrast and are blurred. We propose a nonlinear signal processing technique that significantly improves visual and image qualities (contrast and resolution) of low-contrast infrared images. The proposed method enables the use of infrared cameras for various purposes such as night shot and poor lighting environments under poor lighting conditions.

  4. Adaptive optics improves multiphoton super-resolution imaging

    Science.gov (United States)

    Zheng, Wei; Wu, Yicong; Winter, Peter; Shroff, Hari

    2018-02-01

    Three dimensional (3D) fluorescence microscopy has been essential for biological studies. It allows interrogation of structure and function at spatial scales spanning the macromolecular, cellular, and tissue levels. Critical factors to consider in 3D microscopy include spatial resolution, signal-to-noise (SNR), signal-to-background (SBR), and temporal resolution. Maintaining high quality imaging becomes progressively more difficult at increasing depth (where optical aberrations, induced by inhomogeneities of refractive index in the sample, degrade resolution and SNR), and in thick or densely labeled samples (where out-of-focus background can swamp the valuable, in-focus-signal from each plane). In this report, we introduce our new instrumentation to address these problems. A multiphoton structured illumination microscope was simply modified to integrate an adpative optics system for optical aberrations correction. Firstly, the optical aberrations are determined using direct wavefront sensing with a nonlinear guide star and subsequently corrected using a deformable mirror, restoring super-resolution information. We demonstrate the flexibility of our adaptive optics approach on a variety of semi-transparent samples, including bead phantoms, cultured cells in collagen gels and biological tissues. The performance of our super-resolution microscope is improved in all of these samples, as peak intensity is increased (up to 40-fold) and resolution recovered (up to 176+/-10 nm laterally and 729+/-39 nm axially) at depths up to 250 μm from the coverslip surface.

  5. Resolution enhancement techniques in microscopy

    Science.gov (United States)

    Cremer, Christoph; Masters, Barry R.

    2013-05-01

    We survey the history of resolution enhancement techniques in microscopy and their impact on current research in biomedicine. Often these techniques are labeled superresolution, or enhanced resolution microscopy, or light-optical nanoscopy. First, we introduce the development of diffraction theory in its relation to enhanced resolution; then we explore the foundations of resolution as expounded by the astronomers and the physicists and describe the conditions for which they apply. Then we elucidate Ernst Abbe's theory of optical formation in the microscope, and its experimental verification and dissemination to the world wide microscope communities. Second, we describe and compare the early techniques that can enhance the resolution of the microscope. Third, we present the historical development of various techniques that substantially enhance the optical resolution of the light microscope. These enhanced resolution techniques in their modern form constitute an active area of research with seminal applications in biology and medicine. Our historical survey of the field of resolution enhancement uncovers many examples of reinvention, rediscovery, and independent invention and development of similar proposals, concepts, techniques, and instruments. Attribution of credit is therefore confounded by the fact that for understandable reasons authors stress the achievements from their own research groups and sometimes obfuscate their contributions and the prior art of others. In some cases, attribution of credit is also made more complex by the fact that long term developments are difficult to allocate to a specific individual because of the many mutual connections often existing between sometimes fiercely competing, sometimes strongly collaborating groups. Since applications in biology and medicine have been a major driving force in the development of resolution enhancing approaches, we focus on the contribution of enhanced resolution to these fields.

  6. Simultaneous super-resolution and blind deconvolution

    International Nuclear Information System (INIS)

    Sroubek, F; Flusser, J; Cristobal, G

    2008-01-01

    In many real applications, blur in input low-resolution images is a nuisance, which prevents traditional super-resolution methods from working correctly. This paper presents a unifying approach to the blind deconvolution and superresolution problem of multiple degraded low-resolution frames of the original scene. We introduce a method which assumes no prior information about the shape of degradation blurs and which is properly defined for any rational (fractional) resolution factor. The method minimizes a regularized energy function with respect to the high-resolution image and blurs, where regularization is carried out in both the image and blur domains. The blur regularization is based on a generalized multichannel blind deconvolution constraint. Experiments on real data illustrate robustness and utilization of the method

  7. High-resolution intravital microscopy.

    Directory of Open Access Journals (Sweden)

    Volker Andresen

    Full Text Available Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy--the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and

  8. High-Resolution Intravital Microscopy

    Science.gov (United States)

    Andresen, Volker; Pollok, Karolin; Rinnenthal, Jan-Leo; Oehme, Laura; Günther, Robert; Spiecker, Heinrich; Radbruch, Helena; Gerhard, Jenny; Sporbert, Anje; Cseresnyes, Zoltan; Hauser, Anja E.; Niesner, Raluca

    2012-01-01

    Cellular communication constitutes a fundamental mechanism of life, for instance by permitting transfer of information through synapses in the nervous system and by leading to activation of cells during the course of immune responses. Monitoring cell-cell interactions within living adult organisms is crucial in order to draw conclusions on their behavior with respect to the fate of cells, tissues and organs. Until now, there is no technology available that enables dynamic imaging deep within the tissue of living adult organisms at sub-cellular resolution, i.e. detection at the level of few protein molecules. Here we present a novel approach called multi-beam striped-illumination which applies for the first time the principle and advantages of structured-illumination, spatial modulation of the excitation pattern, to laser-scanning-microscopy. We use this approach in two-photon-microscopy - the most adequate optical deep-tissue imaging-technique. As compared to standard two-photon-microscopy, it achieves significant contrast enhancement and up to 3-fold improved axial resolution (optical sectioning) while photobleaching, photodamage and acquisition speed are similar. Its imaging depth is comparable to multifocal two-photon-microscopy and only slightly less than in standard single-beam two-photon-microscopy. Precisely, our studies within mouse lymph nodes demonstrated 216% improved axial and 23% improved lateral resolutions at a depth of 80 µm below the surface. Thus, we are for the first time able to visualize the dynamic interactions between B cells and immune complex deposits on follicular dendritic cells within germinal centers (GCs) of live mice. These interactions play a decisive role in the process of clonal selection, leading to affinity maturation of the humoral immune response. This novel high-resolution intravital microscopy method has a huge potential for numerous applications in neurosciences, immunology, cancer research and developmental biology

  9. Improved axial resolution of FINCH fluorescence microscopy when combined with spinning disk confocal microscopy.

    Science.gov (United States)

    Siegel, Nisan; Brooker, Gary

    2014-09-22

    FINCH holographic fluorescence microscopy creates super-resolved images with enhanced depth of focus. Addition of a Nipkow disk real-time confocal image scanner is shown to reduce the FINCH depth of focus while improving transverse confocal resolution in a combined method called "CINCH".

  10. Unraveling the Thousand Word Picture: An Introduction to Super-Resolution Data Analysis.

    Science.gov (United States)

    Lee, Antony; Tsekouras, Konstantinos; Calderon, Christopher; Bustamante, Carlos; Pressé, Steve

    2017-06-14

    Super-resolution microscopy provides direct insight into fundamental biological processes occurring at length scales smaller than light's diffraction limit. The analysis of data at such scales has brought statistical and machine learning methods into the mainstream. Here we provide a survey of data analysis methods starting from an overview of basic statistical techniques underlying the analysis of super-resolution and, more broadly, imaging data. We subsequently break down the analysis of super-resolution data into four problems: the localization problem, the counting problem, the linking problem, and what we've termed the interpretation problem.

  11. Assessing resolution in live cell structured illumination microscopy

    Science.gov (United States)

    Pospíšil, Jakub; Fliegel, Karel; Klíma, Miloš

    2017-12-01

    Structured Illumination Microscopy (SIM) is a powerful super-resolution technique, which is able to enhance the resolution of optical microscope beyond the Abbe diffraction limit. In the last decade, numerous SIM methods that achieve the resolution of 100 nm in the lateral dimension have been developed. The SIM setups with new high-speed cameras and illumination pattern generators allow rapid acquisition of the live specimen. Therefore, SIM is widely used for investigation of the live structures in molecular and live cell biology. Quantitative evaluation of resolution enhancement in a real sample is essential to describe the efficiency of super-resolution microscopy technique. However, measuring the resolution of a live cell sample is a challenging task. Based on our experimental findings, the widely used Fourier ring correlation (FRC) method does not seem to be well suited for measuring the resolution of SIM live cell video sequences. Therefore, the resolution assessing methods based on Fourier spectrum analysis are often used. We introduce a measure based on circular average power spectral density (PSDca) estimated from a single SIM image (one video frame). PSDca describes the distribution of the power of a signal with respect to its spatial frequency. Spatial resolution corresponds to the cut-off frequency in Fourier space. In order to estimate the cut-off frequency from a noisy signal, we use a spectral subtraction method for noise suppression. In the future, this resolution assessment approach might prove useful also for single-molecule localization microscopy (SMLM) live cell imaging.

  12. Super-Resolution for Synthetic Zooming

    Directory of Open Access Journals (Sweden)

    Li Xin

    2006-01-01

    Full Text Available Optical zooming is an important feature of imaging systems. In this paper, we investigate a low-cost signal processing alternative to optical zooming—synthetic zooming by super-resolution (SR techniques. Synthetic zooming is achieved by registering a sequence of low-resolution (LR images acquired at varying focal lengths and reconstructing the SR image at a larger focal length or increased spatial resolution. Under the assumptions of constant scene depth and zooming speed, we argue that the motion trajectories of all physical points are related to each other by a unique vanishing point and present a robust technique for estimating its D coordinate. Such a line-geometry-based registration is the foundation of SR for synthetic zooming. We address the issue of data inconsistency arising from the varying focal length of optical lens during the zooming process. To overcome the difficulty of data inconsistency, we propose a two-stage Delaunay-triangulation-based interpolation for fusing the LR image data. We also present a PDE-based nonlinear deblurring to accommodate the blindness and variation of sensor point spread functions. Simulation results with real-world images have verified the effectiveness of the proposed SR techniques for synthetic zooming.

  13. Super-resolution from single photon emission: toward biological application

    Science.gov (United States)

    Moreva, E.; Traina, P.; Forneris, J.; Ditalia Tchernij, S.; Guarina, L.; Franchino, C.; Picollo, F.; Ruo Berchera, I.; Brida, G.; Degiovanni, I. P.; Carabelli, V.; Olivero, P.; Genovese, M.

    2017-08-01

    Properties of quantum light represent a tool for overcoming limits of classical optics. Several experiments have demonstrated this advantage ranging from quantum enhanced imaging to quantum illumination. In this work, experimental demonstration of quantum-enhanced resolution in confocal fluorescence microscopy will be presented. This is achieved by exploiting the non-classical photon statistics of fluorescence emission of single nitrogen-vacancy (NV) color centers in diamond. By developing a general model of super-resolution based on the direct sampling of the kth-order autocorrelation function of the photoluminescence signal, we show the possibility to resolve, in principle, arbitrarily close emitting centers. Finally, possible applications of NV-based fluorescent nanodiamonds in biosensing and future developments will be presented.

  14. Super-resolution fluorescence imaging of membrane nanoscale architectures of hematopoietic stem cell homing and migration molecules

    KAUST Repository

    AbuZineh, Karmen

    2017-01-01

    Recent development of super-resolution (SR) fluorescence microscopy techniques has provided a new tool for direct visualization of subcellular structures and their dynamics in cells. The homing of Hematopoietic stem/progenitor cells (HSPCs) to bone

  15. Super-resolution imaging applied to moving object tracking

    Science.gov (United States)

    Swalaganata, Galandaru; Ratna Sulistyaningrum, Dwi; Setiyono, Budi

    2017-10-01

    Moving object tracking in a video is a method used to detect and analyze changes that occur in an object that being observed. Visual quality and the precision of the tracked target are highly wished in modern tracking system. The fact that the tracked object does not always seem clear causes the tracking result less precise. The reasons are low quality video, system noise, small object, and other factors. In order to improve the precision of the tracked object especially for small object, we propose a two step solution that integrates a super-resolution technique into tracking approach. First step is super-resolution imaging applied into frame sequences. This step was done by cropping the frame in several frame or all of frame. Second step is tracking the result of super-resolution images. Super-resolution image is a technique to obtain high-resolution images from low-resolution images. In this research single frame super-resolution technique is proposed for tracking approach. Single frame super-resolution was a kind of super-resolution that it has the advantage of fast computation time. The method used for tracking is Camshift. The advantages of Camshift was simple calculation based on HSV color that use its histogram for some condition and color of the object varies. The computational complexity and large memory requirements required for the implementation of super-resolution and tracking were reduced and the precision of the tracked target was good. Experiment showed that integrate a super-resolution imaging into tracking technique can track the object precisely with various background, shape changes of the object, and in a good light conditions.

  16. Super-nonlinear fluorescence microscopy for high-contrast deep tissue imaging

    Science.gov (United States)

    Wei, Lu; Zhu, Xinxin; Chen, Zhixing; Min, Wei

    2014-02-01

    Two-photon excited fluorescence microscopy (TPFM) offers the highest penetration depth with subcellular resolution in light microscopy, due to its unique advantage of nonlinear excitation. However, a fundamental imaging-depth limit, accompanied by a vanishing signal-to-background contrast, still exists for TPFM when imaging deep into scattering samples. Formally, the focusing depth, at which the in-focus signal and the out-of-focus background are equal to each other, is defined as the fundamental imaging-depth limit. To go beyond this imaging-depth limit of TPFM, we report a new class of super-nonlinear fluorescence microscopy for high-contrast deep tissue imaging, including multiphoton activation and imaging (MPAI) harnessing novel photo-activatable fluorophores, stimulated emission reduced fluorescence (SERF) microscopy by adding a weak laser beam for stimulated emission, and two-photon induced focal saturation imaging with preferential depletion of ground-state fluorophores at focus. The resulting image contrasts all exhibit a higher-order (third- or fourth- order) nonlinear signal dependence on laser intensity than that in the standard TPFM. Both the physical principles and the imaging demonstrations will be provided for each super-nonlinear microscopy. In all these techniques, the created super-nonlinearity significantly enhances the imaging contrast and concurrently extends the imaging depth-limit of TPFM. Conceptually different from conventional multiphoton processes mediated by virtual states, our strategy constitutes a new class of fluorescence microscopy where high-order nonlinearity is mediated by real population transfer.

  17. Deterministic phase measurements exhibiting super-sensitivity and super-resolution

    DEFF Research Database (Denmark)

    Schäfermeier, Clemens; Ježek, Miroslav; Madsen, Lars S.

    2018-01-01

    Phase super-sensitivity is obtained when the sensitivity in a phase measurement goes beyond the quantum shot noise limit, whereas super-resolution is obtained when the interference fringes in an interferometer are narrower than half the input wavelength. Here we show experimentally that these two...

  18. Improved Interpolation Kernels for Super-resolution Algorithms

    DEFF Research Database (Denmark)

    Rasti, Pejman; Orlova, Olga; Tamberg, Gert

    2016-01-01

    Super resolution (SR) algorithms are widely used in forensics investigations to enhance the resolution of images captured by surveillance cameras. Such algorithms usually use a common interpolation algorithm to generate an initial guess for the desired high resolution (HR) image. This initial guess...... when their original interpolation kernel is replaced by the ones introduced in this work....

  19. Electron microscopy at atomic resolution

    Energy Technology Data Exchange (ETDEWEB)

    Gronsky, R.

    1983-11-01

    The direct imaging of atomic structure in solids has become increasingly easier to accomplish with modern transmission electron microscopes, many of which have an information retrieval limit near 0.2 nm point resolution. Achieving better resolution, particularly with any useful range of specimen tilting, requires a major design effort. This presentation describes the new Atomic Resolution Microscope (ARM), recently put into operation at the Lawrence Berkeley Laboratory. Capable of 0.18 nm or better interpretable resolution over a voltage range of 400 kV to 1000 kV with +- 40/sup 0/ biaxial specimen tilting, the ARM features a number of new electron-optical and microprocessor-control designs. These are highlighted, and its atomic resolution performance demonstrated for a selection of inorganic crystals.

  20. Electron microscopy at atomic resolution

    International Nuclear Information System (INIS)

    Gronsky, R.

    1983-11-01

    The direct imaging of atomic structure in solids has become increasingly easier to accomplish with modern transmission electron microscopes, many of which have an information retrieval limit near 0.2 nm point resolution. Achieving better resolution, particularly with any useful range of specimen tilting, requires a major design effort. This presentation describes the new Atomic Resolution Microscope (ARM), recently put into operation at the Lawrence Berkeley Laboratory. Capable of 0.18 nm or better interpretable resolution over a voltage range of 400 kV to 1000 kV with +- 40 0 biaxial specimen tilting, the ARM features a number of new electron-optical and microprocessor-control designs. These are highlighted, and its atomic resolution performance demonstrated for a selection of inorganic crystals

  1. Super-resolution biomolecular crystallography with low-resolution data.

    Science.gov (United States)

    Schröder, Gunnar F; Levitt, Michael; Brunger, Axel T

    2010-04-22

    -ray crystallography and cryo-electron microscopy: as optical imaging advances to subnanometre resolution, it can use similar tools.

  2. Super-resolution structure of DNA significantly differs in buccal cells of controls and Alzheimer's patients.

    Science.gov (United States)

    Garcia, Angeles; Huang, David; Righolt, Amanda; Righolt, Christiaan; Kalaw, Maria Carmela; Mathur, Shubha; McAvoy, Elizabeth; Anderson, James; Luedke, Angela; Itorralba, Justine; Mai, Sabine

    2017-09-01

    The advent of super-resolution microscopy allowed for new insights into cellular and physiological processes of normal and diseased cells. In this study, we report for the first time on the super-resolved DNA structure of buccal cells from patients with Alzheimer's disease (AD) versus age- and gender-matched healthy, non-caregiver controls. In this super-resolution study cohort of 74 participants, buccal cells were collected and their spatial DNA organization in the nucleus examined by 3D Structured Illumination Microscopy (3D-SIM). Quantitation of the super-resolution DNA structure revealed that the nuclear super-resolution DNA structure of individuals with AD significantly differs from that of their controls (p structure of AD significantly differs in mild, moderate, and severe disease with respect to the DNA-containing and DNA-free/poor spaces. We conclude that whole genome remodeling is a feature of buccal cells in AD. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

  3. High Resolution Scanning Ion Microscopy

    NARCIS (Netherlands)

    Castaldo, V.

    2011-01-01

    The structure of the thesis is the following. The first chapter is an introduction to scanning microscopy, where the path that led to the Focused Ion Beam (FIB) is described and the main differences between electrons and ion beams are highlighted. Chapter 2 is what is normally referred to (which I

  4. Live-cell super-resolution imaging of intrinsically fast moving flagellates

    International Nuclear Information System (INIS)

    Glogger, M; Subota, I; Spindler, M-C; Engstler, M; Fenz, S F; Stichler, S; Bertlein, S; Teßmar, J; Groll, J

    2017-01-01

    Recent developments in super-resolution microscopy make it possible to resolve structures in biological cells at a spatial resolution of a few nm and observe dynamical processes with a temporal resolution of ms to μ s. However, the optimal structural resolution requires repeated illumination cycles and is thus limited to chemically fixed cells. For live cell applications substantial improvement over classical Abbe-limited imaging can already be obtained in adherent or slow moving cells. Nonetheless, a large group of cells are fast moving and thus could not yet be addressed with live cell super-resolution microscopy. These include flagellate pathogens like African trypanosomes, the causative agents of sleeping sickness in humans and nagana in livestock. Here, we present an embedding method based on a in situ forming cytocompatible UV-crosslinked hydrogel. The fast cross-linking hydrogel immobilizes trypanosomes efficiently to allow microscopy on the nanoscale. We characterized both the trypanosomes and the hydrogel with respect to their autofluorescence properties and found them suitable for single-molecule fluorescence microscopy (SMFM). As a proof of principle, SMFM was applied to super-resolve a structure inside the living trypanosome. We present an image of a flagellar axoneme component recorded by using the intrinsic blinking behavior of eYFP. (paper)

  5. Super-Resolution for Color Imagery

    Science.gov (United States)

    2017-09-01

    September 2017 2. REPORT TYPE Technical Report 3. DATES COVERED (From - To) 1 October 2016‒30 September 2017 4. TITLE AND SUBTITLE Super...slightly lighter shade of purple. There is slightly less purple coloring on the white paper than with the other 2 color spaces, and the black font is

  6. Resolution improvement by nonconfocal theta microscopy.

    Science.gov (United States)

    Lindek, S; Stelzer, E H

    1999-11-01

    We present a novel scanning fluorescence microscopy technique, nonconfocal theta microscopy (NCTM), that provides almost isotropic resolution. In NCTM, multiphoton absorption from two orthogonal illumination directions is used to induce fluorescence emission. Therefore the point-spread function of the microscope is described by the product of illumination point-spread functions with reduced spatial overlap, which provides the resolution improvement and the more isotropic observation volume. We discuss the technical details of this new method.

  7. Super-resolution for asymmetric resolution of FIB-SEM 3D imaging using AI with deep learning.

    Science.gov (United States)

    Hagita, Katsumi; Higuchi, Takeshi; Jinnai, Hiroshi

    2018-04-12

    Scanning electron microscopy equipped with a focused ion beam (FIB-SEM) is a promising three-dimensional (3D) imaging technique for nano- and meso-scale morphologies. In FIB-SEM, the specimen surface is stripped by an ion beam and imaged by an SEM installed orthogonally to the FIB. The lateral resolution is governed by the SEM, while the depth resolution, i.e., the FIB milling direction, is determined by the thickness of the stripped thin layer. In most cases, the lateral resolution is superior to the depth resolution; hence, asymmetric resolution is generated in the 3D image. Here, we propose a new approach based on an image-processing or deep-learning-based method for super-resolution of 3D images with such asymmetric resolution, so as to restore the depth resolution to achieve symmetric resolution. The deep-learning-based method learns from high-resolution sub-images obtained via SEM and recovers low-resolution sub-images parallel to the FIB milling direction. The 3D morphologies of polymeric nano-composites are used as test images, which are subjected to the deep-learning-based method as well as conventional methods. We find that the former yields superior restoration, particularly as the asymmetric resolution is increased. Our super-resolution approach for images having asymmetric resolution enables observation time reduction.

  8. Oblique reconstructions in tomosynthesis. II. Super-resolution

    International Nuclear Information System (INIS)

    Acciavatti, Raymond J.; Maidment, Andrew D. A.

    2013-01-01

    Purpose: In tomosynthesis, super-resolution has been demonstrated using reconstruction planes parallel to the detector. Super-resolution allows for subpixel resolution relative to the detector. The purpose of this work is to develop an analytical model that generalizes super-resolution to oblique reconstruction planes.Methods: In a digital tomosynthesis system, a sinusoidal test object is modeled along oblique angles (i.e., “pitches”) relative to the plane of the detector in a 3D divergent-beam acquisition geometry. To investigate the potential for super-resolution, the input frequency is specified to be greater than the alias frequency of the detector. Reconstructions are evaluated in an oblique plane along the extent of the object using simple backprojection (SBP) and filtered backprojection (FBP). By comparing the amplitude of the reconstruction against the attenuation coefficient of the object at various frequencies, the modulation transfer function (MTF) is calculated to determine whether modulation is within detectable limits for super-resolution. For experimental validation of super-resolution, a goniometry stand was used to orient a bar pattern phantom along various pitches relative to the breast support in a commercial digital breast tomosynthesis system.Results: Using theoretical modeling, it is shown that a single projection image cannot resolve a sine input whose frequency exceeds the detector alias frequency. The high frequency input is correctly visualized in SBP or FBP reconstruction using a slice along the pitch of the object. The Fourier transform of this reconstructed slice is maximized at the input frequency as proof that the object is resolved. Consistent with the theoretical results, experimental images of a bar pattern phantom showed super-resolution in oblique reconstructions. At various pitches, the highest frequency with detectable modulation was determined by visual inspection of the bar patterns. The dependency of the highest

  9. Oblique reconstructions in tomosynthesis. II. Super-resolution

    Science.gov (United States)

    Acciavatti, Raymond J.; Maidment, Andrew D. A.

    2013-01-01

    Purpose: In tomosynthesis, super-resolution has been demonstrated using reconstruction planes parallel to the detector. Super-resolution allows for subpixel resolution relative to the detector. The purpose of this work is to develop an analytical model that generalizes super-resolution to oblique reconstruction planes. Methods: In a digital tomosynthesis system, a sinusoidal test object is modeled along oblique angles (i.e., “pitches”) relative to the plane of the detector in a 3D divergent-beam acquisition geometry. To investigate the potential for super-resolution, the input frequency is specified to be greater than the alias frequency of the detector. Reconstructions are evaluated in an oblique plane along the extent of the object using simple backprojection (SBP) and filtered backprojection (FBP). By comparing the amplitude of the reconstruction against the attenuation coefficient of the object at various frequencies, the modulation transfer function (MTF) is calculated to determine whether modulation is within detectable limits for super-resolution. For experimental validation of super-resolution, a goniometry stand was used to orient a bar pattern phantom along various pitches relative to the breast support in a commercial digital breast tomosynthesis system. Results: Using theoretical modeling, it is shown that a single projection image cannot resolve a sine input whose frequency exceeds the detector alias frequency. The high frequency input is correctly visualized in SBP or FBP reconstruction using a slice along the pitch of the object. The Fourier transform of this reconstructed slice is maximized at the input frequency as proof that the object is resolved. Consistent with the theoretical results, experimental images of a bar pattern phantom showed super-resolution in oblique reconstructions. At various pitches, the highest frequency with detectable modulation was determined by visual inspection of the bar patterns. The dependency of the highest

  10. Confocal pore size measurement based on super-resolution image restoration.

    Science.gov (United States)

    Liu, Dali; Wang, Yun; Qiu, Lirong; Mao, Xinyue; Zhao, Weiqian

    2014-09-01

    A confocal pore size measurement based on super-resolution image restoration is proposed to obtain a fast and accurate measurement for submicrometer pore size of nuclear track-etched membranes (NTEMs). This method facilitates the online inspection of the pore size evolution during etching. Combining confocal microscopy with super-resolution image restoration significantly improves the lateral resolution of the NTEM image, yields a reasonable circle edge-setting criterion of 0.2408, and achieves precise pore edge detection. Theoretical analysis shows that the minimum measuring diameter can reach 0.19 μm, and the root mean square of the residuals is only 1.4 nm. Edge response simulation and experiment reveal that the edge response of the proposed method is better than 80 nm. The NTEM pore size measurement results obtained by the proposed method agree well with that obtained by scanning electron microscopy.

  11. Localization-based super-resolution imaging meets high-content screening.

    Science.gov (United States)

    Beghin, Anne; Kechkar, Adel; Butler, Corey; Levet, Florian; Cabillic, Marine; Rossier, Olivier; Giannone, Gregory; Galland, Rémi; Choquet, Daniel; Sibarita, Jean-Baptiste

    2017-12-01

    Single-molecule localization microscopy techniques have proven to be essential tools for quantitatively monitoring biological processes at unprecedented spatial resolution. However, these techniques are very low throughput and are not yet compatible with fully automated, multiparametric cellular assays. This shortcoming is primarily due to the huge amount of data generated during imaging and the lack of software for automation and dedicated data mining. We describe an automated quantitative single-molecule-based super-resolution methodology that operates in standard multiwell plates and uses analysis based on high-content screening and data-mining software. The workflow is compatible with fixed- and live-cell imaging and allows extraction of quantitative data like fluorophore photophysics, protein clustering or dynamic behavior of biomolecules. We demonstrate that the method is compatible with high-content screening using 3D dSTORM and DNA-PAINT based super-resolution microscopy as well as single-particle tracking.

  12. Extreme super-resolution using the spherical geodesic waveguide

    Science.gov (United States)

    Miñano, Juan Carlos; González, Juan Carlos; Benítez, Pablo; Grabovičkić, Dejan

    2012-06-01

    Leonhardt demonstrated (2009) that the 2D Maxwell Fish Eye lens (MFE) can focus perfectly 2D Helmholtz waves of arbitrary frequency, i.e., it can transport perfectly an outward (monopole) 2D Helmholtz wave field, generated by a point source, towards a "perfect point drain" located at the corresponding image point. Moreover, a prototype with λ/5 super-resolution (SR) property for one microwave frequency has been manufactured and tested (Ma et al, 2010). Although this prototype has been loaded with an impedance different from the "perfect point drain", it has shown super-resolution property. However, neither software simulations nor experimental measurements for a broad band of frequencies have yet been reported. Here we present steady state simulations for two cases, using perfect drain as suggested by Leonhardt and without perfect drain as in the prototype. All the simulations have been done using a device equivalent to the MFE, called the Spherical Geodesic Waveguide (SGW). The results show the super-resolution up to λ/3000, for the system loaded with the perfect drain, and up to λ /500 for a not perfect load. In both cases super-resolution only happens for discrete number of frequencies. Out of these frequencies, the SGW does not show super-resolution in the analysis carried out.

  13. High resolution eddy current microscopy

    Science.gov (United States)

    Lantz, M. A.; Jarvis, S. P.; Tokumoto, H.

    2001-01-01

    We describe a sensitive scanning force microscope based technique for measuring local variations in resistivity by monitoring changes in the eddy current induced damping of a cantilever with a magnetic tip oscillating above a conducting sample. To achieve a high sensitivity, we used a cantilever with an FeNdBLa particle mounted on the tip. Resistivity measurements are demonstrated on a silicon test structure with a staircase doping profile. Regions with resistivities of 0.0013, 0.0041, and 0.022 Ω cm are clearly resolved with a lateral resolution of approximately 180 nm. For this range of resistivities, the eddy current induced damping is found to depend linearly on the sample resistivity.

  14. Nonlinear super-resolution nano-optics and applications

    CERN Document Server

    Wei, Jingsong

    2015-01-01

    This book covers many advances in the subjects of nano-optics and nano photonics. The author describes the principle and technical schematics of common methods for breaking through the optical diffraction limit and focuses on realizing optical super-resolution with nonlinear effects of thin film materials. The applications of nonlinear optical super-resolution effects in nano-data storage, nanolithography, and nano-imaging are also presented. This book is useful to graduate students majoring in optics and nano science and also serves as a reference book for academic researchers, engineers, technical professionals in the fields of super-resolution optics and laser techniques, nano-optics and nano photonics, nano-data storage, nano imaging, micro/nanofabrication and nanolithography and nonlinear optics.

  15. Super resolution imaging of genetically labelled synapses in Drosophila brain tissue

    Directory of Open Access Journals (Sweden)

    Isabelle Ayumi Spühler

    2016-05-01

    Full Text Available Understanding synaptic connectivity and plasticity within brain circuits and their relationship to learning and behavior is a fundamental quest in neuroscience. Visualizing the fine details of synapses using optical microscopy remains however a major technical challenge. Super resolution microscopy opens the possibility to reveal molecular features of synapses beyond the diffraction limit. With direct stochastic optical reconstruction microscopy, dSTORM, we image synaptic proteins in the brain tissue of the fruit fly, Drosophila melanogaster. Super resolution imaging of brain tissue harbors difficulties due to light scattering and the density of signals. In order to reduce out of focus signal, we take advantage of the genetic tools available in the Drosophila and have fluorescently tagged synaptic proteins expressed in only a small number of neurons. These neurons form synapses within the calyx of the mushroom body, a distinct brain region involved in associative memory formation. Our results show that super resolution microscopy, in combination with genetically labelled synaptic proteins, is a powerful tool to investigate synapses in a quantitative fashion providing an entry point for studies on synaptic plasticity during learning and memory formation

  16. Super Resolution Imaging of Genetically Labeled Synapses in Drosophila Brain Tissue.

    Science.gov (United States)

    Spühler, Isabelle A; Conley, Gaurasundar M; Scheffold, Frank; Sprecher, Simon G

    2016-01-01

    Understanding synaptic connectivity and plasticity within brain circuits and their relationship to learning and behavior is a fundamental quest in neuroscience. Visualizing the fine details of synapses using optical microscopy remains however a major technical challenge. Super resolution microscopy opens the possibility to reveal molecular features of synapses beyond the diffraction limit. With direct stochastic optical reconstruction microscopy, dSTORM, we image synaptic proteins in the brain tissue of the fruit fly, Drosophila melanogaster. Super resolution imaging of brain tissue harbors difficulties due to light scattering and the density of signals. In order to reduce out of focus signal, we take advantage of the genetic tools available in the Drosophila and have fluorescently tagged synaptic proteins expressed in only a small number of neurons. These neurons form synapses within the calyx of the mushroom body, a distinct brain region involved in associative memory formation. Our results show that super resolution microscopy, in combination with genetically labeled synaptic proteins, is a powerful tool to investigate synapses in a quantitative fashion providing an entry point for studies on synaptic plasticity during learning and memory formation.

  17. Finite detector based projection model for super resolution CT

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Hengyong; Wang, Ge [Wake Forest Univ. Health Sciences, Winston-Salem, NC (United States). Dept. of Radiology; Virgina Tech, Blacksburg, VA (United States). Biomedical Imaging Div.

    2011-07-01

    For finite detector and focal spot sizes, here we propose a projection model for super resolution CT. First, for a given X-ray source point, a projection datum is modeled as an area integral over a narrow fan-beam connecting the detector elemental borders and the X-ray source point. Then, the final projection value is expressed as the integral obtained in the first step over the whole focal spot support. An ordered-subset simultaneous algebraic reconstruction technique (OS-SART) is developed using the proposed projection model. In the numerical simulation, our method produces super spatial resolution and suppresses high-frequency artifacts. (orig.)

  18. Application of Super-Resolution Image Reconstruction to Digital Holography

    Directory of Open Access Journals (Sweden)

    Zhang Shuqun

    2006-01-01

    Full Text Available We describe a new application of super-resolution image reconstruction to digital holography which is a technique for three-dimensional information recording and reconstruction. Digital holography has suffered from the low resolution of CCD sensors, which significantly limits the size of objects that can be recorded. The existing solution to this problem is to use optics to bandlimit the object to be recorded, which can cause the loss of details. Here super-resolution image reconstruction is proposed to be applied in enhancing the spatial resolution of digital holograms. By introducing a global camera translation before sampling, a high-resolution hologram can be reconstructed from a set of undersampled hologram images. This permits the recording of larger objects and reduces the distance between the object and the hologram. Practical results from real and simulated holograms are presented to demonstrate the feasibility of the proposed technique.

  19. Super-resolution for scanning light stimulation systems

    Energy Technology Data Exchange (ETDEWEB)

    Bitzer, L. A.; Neumann, K.; Benson, N., E-mail: niels.benson@uni-due.de; Schmechel, R. [Faculty of Engineering, NST and CENIDE, University of Duisburg-Essen, Bismarckstr. 81, 47057 Duisburg (Germany)

    2016-09-15

    Super-resolution (SR) is a technique used in digital image processing to overcome the resolution limitation of imaging systems. In this process, a single high resolution image is reconstructed from multiple low resolution images. SR is commonly used for CCD and CMOS (Complementary Metal-Oxide-Semiconductor) sensor images, as well as for medical applications, e.g., magnetic resonance imaging. Here, we demonstrate that super-resolution can be applied with scanning light stimulation (LS) systems, which are common to obtain space-resolved electro-optical parameters of a sample. For our purposes, the Projection Onto Convex Sets (POCS) was chosen and modified to suit the needs of LS systems. To demonstrate the SR adaption, an Optical Beam Induced Current (OBIC) LS system was used. The POCS algorithm was optimized by means of OBIC short circuit current measurements on a multicrystalline solar cell, resulting in a mean square error reduction of up to 61% and improved image quality.

  20. Far-field super-resolution imaging of resonant multiples

    KAUST Repository

    Guo, Bowen

    2016-05-20

    We demonstrate for the first time that seismic resonant multiples, usually considered as noise, can be used for super-resolution imaging in the far-field region of sources and receivers. Tests with both synthetic data and field data show that resonant multiples can image reflector boundaries with resolutions more than twice the classical resolution limit. Resolution increases with the order of the resonant multiples. This procedure has important applications in earthquake and exploration seismology, radar, sonar, LIDAR (light detection and ranging), and ultrasound imaging, where the multiples can be used to make high-resolution images.

  1. Comparison of super-resolution benefits for downsampled iages and real low-resolution data

    NARCIS (Netherlands)

    Peng, Y.; Spreeuwers, Lieuwe Jan; Gökberk, B.; Veldhuis, Raymond N.J.

    2013-01-01

    Recently, more and more researchers are exploring the benefits of super-resolution methods on low-resolution face recognition. However, often results presented are obtained on downsampled high-resolution face images. Because downsampled images are different from real images taken at low resolution,

  2. SINGLE FRAME SUPER RESOLUTION OF NONCOOPERATIVE IRIS IMAGES

    Directory of Open Access Journals (Sweden)

    Anand Deshpande

    2016-11-01

    Full Text Available Image super-resolution, a process to enhance image resolution, has important applications in biometrics, satellite imaging, high definition television, medical imaging, etc. The long range captured iris identification systems often suffer from low resolution and meager focus of the captured iris images. These degrade the iris recognition performance. This paper proposes enhanced iterated back projection (EIBP method to super resolute the long range captured iris polar images. The performance of proposed method is tested and analyzed on CASIA long range iris database by comparing peak signal to noise ratio (PSNR and structural similarity index (SSIM with state-of-the-art super resolution (SR algorithms. It is further analyzed by increasing the up-sampling factor. Performance analysis shows that the proposed method is superior to state-of-the-art algorithms, the peak signal-to-noise ratio improved about 0.1-1.5 dB. The results demonstrate that the proposed method is well suited to super resolve the iris polar images captured at a long distance

  3. Deep Learning based Super-Resolution for Improved Action Recognition

    DEFF Research Database (Denmark)

    Nasrollahi, Kamal; Guerrero, Sergio Escalera; Rasti, Pejman

    2015-01-01

    with results of a state-of- the-art deep learning-based super-resolution algorithm, through an alpha-blending approach. The experimental results obtained on down-sampled version of a large subset of Hoolywood2 benchmark database show the importance of the proposed system in increasing the recognition rate...

  4. Video super-resolution using simultaneous motion and intensity calculations

    DEFF Research Database (Denmark)

    Keller, Sune Høgild; Lauze, Francois Bernard; Nielsen, Mads

    2011-01-01

    for the joint estimation of a super-resolution sequence and its flow field. Via the calculus of variations, this leads to a coupled system of partial differential equations for image sequence and motion estimation. We solve a simplified form of this system and as a by-product we indeed provide a motion field...

  5. Generation of super-resolution stills from video

    CSIR Research Space (South Africa)

    Duvenhage, B

    2014-11-01

    Full Text Available plane. If one accurately registers the image of the target on the focal plane to some reference then one can increase the effective sensor pixel density by stacking or appropriately combining the registered images. The super-resolution technique operates...

  6. Atomic Resolution Microscopy of Nitrides in Steel

    DEFF Research Database (Denmark)

    Danielsen, Hilmar Kjartansson

    2014-01-01

    MN and CrMN type nitride precipitates in 12%Cr steels have been investigated using atomic resolution microscopy. The MN type nitrides were observed to transform into CrMN both by composition and crystallography as Cr diffuses from the matrix into the MN precipitates. Thus a change from one...

  7. Super-resolution optics for virtual reality

    Science.gov (United States)

    Grabovičkić, Dejan; Benitez, Pablo; Miñano, Juan C.; Zamora, Pablo; Buljan, Marina; Narasimhan, Bharathwaj; Nikolic, Milena I.; Lopez, Jesus; Gorospe, Jorge; Sanchez, Eduardo; Lastres, Carmen; Mohedano, Ruben

    2017-06-01

    In present commercial Virtual Reality (VR) headsets the resolution perceived is still limited, since the VR pixel density (typically 10-15 pixels/deg) is well below what the human eye can resolve (60 pixels/deg). We present here novel advanced optical design approaches that dramatically increase the perceived resolution of the VR keeping the large FoV required in VR applications. This approach can be applied to a vast number of optical architectures, including some advanced configurations, as multichannel designs. All this is done at the optical design stage, and no eye tracker is needed in the headset.

  8. Controlled power delivery for super-resolution imaging of biological samples using digital micromirror device

    Science.gov (United States)

    Valiya Peedikakkal, Liyana; Cadby, Ashley

    2017-02-01

    Localization based super resolution images of a biological sample is generally achieved by using high power laser illumination with long exposure time which unfortunately increases photo-toxicity of a sample, making super resolution microscopy, in general, incompatible with live cell imaging. Furthermore, the limitation of photobleaching reduces the ability to acquire time lapse images of live biological cells using fluorescence microscopy. Digital Light Processing (DLP) technology can deliver light at grey scale levels by flickering digital micromirrors at around 290 Hz enabling highly controlled power delivery to samples. In this work, Digital Micromirror Device (DMD) is implemented in an inverse Schiefspiegler telescope setup to control the power and pattern of illumination for super resolution microscopy. We can achieve spatial and temporal patterning of illumination by controlling the DMD pixel by pixel. The DMD allows us to control the power and spatial extent of the laser illumination. We have used this to show that we can reduce the power delivered to the sample to allow for longer time imaging in one area while achieving sub-diffraction STORM imaging in another using higher power densities.

  9. High resolution, high speed ultrahigh vacuum microscopy

    International Nuclear Information System (INIS)

    Poppa, Helmut

    2004-01-01

    The history and future of transmission electron microscopy (TEM) is discussed as it refers to the eventual development of instruments and techniques applicable to the real time in situ investigation of surface processes with high resolution. To reach this objective, it was necessary to transform conventional high resolution instruments so that an ultrahigh vacuum (UHV) environment at the sample site was created, that access to the sample by various in situ sample modification procedures was provided, and that in situ sample exchanges with other integrated surface analytical systems became possible. Furthermore, high resolution image acquisition systems had to be developed to take advantage of the high speed imaging capabilities of projection imaging microscopes. These changes to conventional electron microscopy and its uses were slowly realized in a few international laboratories over a period of almost 40 years by a relatively small number of researchers crucially interested in advancing the state of the art of electron microscopy and its applications to diverse areas of interest; often concentrating on the nucleation, growth, and properties of thin films on well defined material surfaces. A part of this review is dedicated to the recognition of the major contributions to surface and thin film science by these pioneers. Finally, some of the important current developments in aberration corrected electron optics and eventual adaptations to in situ UHV microscopy are discussed. As a result of all the path breaking developments that have led to today's highly sophisticated UHV-TEM systems, integrated fundamental studies are now possible that combine many traditional surface science approaches. Combined investigations to date have involved in situ and ex situ surface microscopies such as scanning tunneling microscopy/atomic force microscopy, scanning Auger microscopy, and photoemission electron microscopy, and area-integrating techniques such as x-ray photoelectron

  10. New device based on the super spatial resolution (SSR) method

    International Nuclear Information System (INIS)

    Soluri, A.; Atzeni, G.; Ucci, A.; Bellone, T.; Cusanno, F.; Rodilossi, G.; Massari, R.

    2013-01-01

    Recently it have been described that innovative methods, namely Super Spatial Resolution (SSR), can be used to improve the scintigraphic imaging. The aim of SSR techniques is the enhancement of the resolution of an imaging system, using information from several images. In this paper we describe a new experimental apparatus that could be used for molecular imaging and small animal imaging. In fact we present a new device, completely automated, that uses the SSR method and provides images with better spatial resolution in comparison to the original resolution. Preliminary small animal imaging studies confirm the feasibility of a very high resolution system in scintigraphic imaging and the possibility to have gamma cameras using the SSR method, to perform the applications on functional imaging. -- Highlights: • Super spatial resolution brings a high resolution image from scintigraphic images. • Resolution improvement depends on the signal to noise ratio of the original images. • The SSR shows significant improvement on spatial resolution in scintigraphic images. • The SSR method is potentially utilizable for all scintigraphic devices

  11. Robust microbubble tracking for super resolution imaging in ultrasound

    DEFF Research Database (Denmark)

    Hansen, Kristoffer B.; Villagómez Hoyos, Carlos Armando; Brasen, Jens Christian

    2016-01-01

    Currently ultrasound resolution is limited by diffraction to approximately half the wavelength of the sound wave employed. In recent years, super resolution imaging techniques have overcome the diffraction limit through the localization and tracking of a sparse set of microbubbles through...... the vasculature. However, this has only been performed on fixated tissue, limiting its clinical application. This paper proposes a technique for making super resolution images on non-fixated tissue by first compensating for tissue movement and then tracking the individual microbubbles. The experiment is performed...... on the kidney of a anesthetized Sprage-Dawley rat by infusing SonoVue at 0.1× original concentration. The algorithm demonstrated in vivo that the motion compensation was capable of removing the movement caused by the mechanical ventilator. The results shows that microbubbles were localized with a higher...

  12. Underwater video enhancement using multi-camera super-resolution

    Science.gov (United States)

    Quevedo, E.; Delory, E.; Callicó, G. M.; Tobajas, F.; Sarmiento, R.

    2017-12-01

    Image spatial resolution is critical in several fields such as medicine, communications or satellite, and underwater applications. While a large variety of techniques for image restoration and enhancement has been proposed in the literature, this paper focuses on a novel Super-Resolution fusion algorithm based on a Multi-Camera environment that permits to enhance the quality of underwater video sequences without significantly increasing computation. In order to compare the quality enhancement, two objective quality metrics have been used: PSNR (Peak Signal-to-Noise Ratio) and the SSIM (Structural SIMilarity) index. Results have shown that the proposed method enhances the objective quality of several underwater sequences, avoiding the appearance of undesirable artifacts, with respect to basic fusion Super-Resolution algorithms.

  13. Live-cell super-resolution imaging of intrinsically fast moving flagellates

    Science.gov (United States)

    Glogger, M.; Stichler, S.; Subota, I.; Bertlein, S.; Spindler, M.-C.; Teßmar, J.; Groll, J.; Engstler, M.; Fenz, S. F.

    2017-02-01

    Recent developments in super-resolution microscopy make it possible to resolve structures in biological cells at a spatial resolution of a few nm and observe dynamical processes with a temporal resolution of ms to μs. However, the optimal structural resolution requires repeated illumination cycles and is thus limited to chemically fixed cells. For live cell applications substantial improvement over classical Abbe-limited imaging can already be obtained in adherent or slow moving cells. Nonetheless, a large group of cells are fast moving and thus could not yet be addressed with live cell super-resolution microscopy. These include flagellate pathogens like African trypanosomes, the causative agents of sleeping sickness in humans and nagana in livestock. Here, we present an embedding method based on a in situ forming cytocompatible UV-crosslinked hydrogel. The fast cross-linking hydrogel immobilizes trypanosomes efficiently to allow microscopy on the nanoscale. We characterized both the trypanosomes and the hydrogel with respect to their autofluorescence properties and found them suitable for single-molecule fluorescence microscopy (SMFM). As a proof of principle, SMFM was applied to super-resolve a structure inside the living trypanosome. We present an image of a flagellar axoneme component recorded by using the intrinsic blinking behavior of eYFP. , which features invited work from the best early-career researchers working within the scope of J Phys D. This project is part of the Journal of Physics series’ 50th anniversary celebrations in 2017. Susanne Fenz was selected by the Editorial Board of J Phys D as an Emerging Talent/Leader.

  14. Identification and super-resolution imaging of ligand-activated receptor dimers in live cells

    Science.gov (United States)

    Winckler, Pascale; Lartigue, Lydia; Giannone, Gregory; de Giorgi, Francesca; Ichas, François; Sibarita, Jean-Baptiste; Lounis, Brahim; Cognet, Laurent

    2013-08-01

    Molecular interactions are key to many chemical and biological processes like protein function. In many signaling processes they occur in sub-cellular areas displaying nanoscale organizations and involving molecular assemblies. The nanometric dimensions and the dynamic nature of the interactions make their investigations complex in live cells. While super-resolution fluorescence microscopies offer live-cell molecular imaging with sub-wavelength resolutions, they lack specificity for distinguishing interacting molecule populations. Here we combine super-resolution microscopy and single-molecule Förster Resonance Energy Transfer (FRET) to identify dimers of receptors induced by ligand binding and provide super-resolved images of their membrane distribution in live cells. By developing a two-color universal-Point-Accumulation-In-the-Nanoscale-Topography (uPAINT) method, dimers of epidermal growth factor receptors (EGFR) activated by EGF are studied at ultra-high densities, revealing preferential cell-edge sub-localization. This methodology which is specifically devoted to the study of molecules in interaction, may find other applications in biological systems where understanding of molecular organization is crucial.

  15. Dynamic placement of plasmonic hotspots for super-resolution surface-enhanced Raman scattering.

    Science.gov (United States)

    Ertsgaard, Christopher T; McKoskey, Rachel M; Rich, Isabel S; Lindquist, Nathan C

    2014-10-28

    In this paper, we demonstrate dynamic placement of locally enhanced plasmonic fields using holographic laser illumination of a silver nanohole array. To visualize these focused "hotspots", the silver surface was coated with various biological samples for surface-enhanced Raman spectroscopy (SERS) imaging. Due to the large field enhancements, blinking behavior of the SERS hotspots was observed and processed using a stochastic optical reconstruction microscopy algorithm enabling super-resolution localization of the hotspots to within 10 nm. These hotspots were then shifted across the surface in subwavelength (hotspots. Using this technique, we also show that such subwavelength shifting and localization of plasmonic hotspots has potential for imaging applications. Interestingly, illuminating the surface with randomly shifting SERS hotspots was sufficient to completely fill in a wide field of view for super-resolution chemical imaging.

  16. Fabrication of optical multilayer for two-color phase plate in super-resolution microscope.

    Science.gov (United States)

    Iketaki, Yoshinori; Kitagawa, Katsuichi; Hidaka, Kohjiro; Kato, Naoki; Hirabayashi, Akira; Bokor, Nandor

    2014-07-01

    In super-resolution microscopy based on fluorescence depletion, the two-color phase plate (TPP) is an indispensable optical element, which can independently control the phase shifts for two beams of different color, i.e., the pump and erase beams. By controlling a phase shift of the erase beam through the TPP, the erase beam can be modulated into a doughnut shape, while the pump beam maintains the initial Gaussian shape. To obtain a reliable optical multiplayer (ML) for the TPP, we designed a ML with only two optical layers by performing numerical optimization. The measured phase shifts generated by the fabricated ML using interferometry correspond to the design values. The beam profiles in the focal plane are also consistent with theoretical results. Although the fabricated ML consists of only two optical layers, the ML can provide a suitable phase modulation function for the TPP in a practical super-resolution microscope.

  17. Fabrication of optical multilayer for two-color phase plate in super-resolution microscope

    International Nuclear Information System (INIS)

    Iketaki, Yoshinori; Kitagawa, Katsuichi; Hidaka, Kohjiro; Kato, Naoki; Hirabayashi, Akira; Bokor, Nandor

    2014-01-01

    In super-resolution microscopy based on fluorescence depletion, the two-color phase plate (TPP) is an indispensable optical element, which can independently control the phase shifts for two beams of different color, i.e., the pump and erase beams. By controlling a phase shift of the erase beam through the TPP, the erase beam can be modulated into a doughnut shape, while the pump beam maintains the initial Gaussian shape. To obtain a reliable optical multiplayer (ML) for the TPP, we designed a ML with only two optical layers by performing numerical optimization. The measured phase shifts generated by the fabricated ML using interferometry correspond to the design values. The beam profiles in the focal plane are also consistent with theoretical results. Although the fabricated ML consists of only two optical layers, the ML can provide a suitable phase modulation function for the TPP in a practical super-resolution microscope

  18. Super-resolution thermographic imaging using blind structured illumination

    Science.gov (United States)

    Burgholzer, Peter; Berer, Thomas; Gruber, Jürgen; Mayr, Günther

    2017-07-01

    Using an infrared camera for thermographic imaging allows the contactless temperature measurement of many surface pixels simultaneously. From the measured surface data, the structure below the surface, embedded inside a sample or tissue, can be reconstructed and imaged, if heated by an excitation light pulse. The main drawback in active thermographic imaging is the degradation of the spatial resolution with the imaging depth, which results in blurred images for deeper lying structures. We circumvent this degradation by using blind structured illumination combined with a non-linear joint sparsity reconstruction algorithm. We demonstrate imaging of a line pattern and a star-shaped structure through a 3 mm thick steel sheet with a resolution four times better than the width of the thermal point-spread-function. The structured illumination is realized by parallel slits cut in an aluminum foil, where the excitation coming from a flashlight can penetrate. This realization of super-resolution thermographic imaging demonstrates that blind structured illumination allows thermographic imaging without high degradation of the spatial resolution for deeper lying structures. The groundbreaking concept of super-resolution can be transferred from optics to diffusive imaging by defining a thermal point-spread-function, which gives the principle resolution limit for a certain signal-to-noise ratio, similar to the Abbe limit for a certain optical wavelength. In future work, the unknown illumination pattern could be the speckle pattern generated by a short laser pulse inside a light scattering sample or tissue.

  19. Texton-based super-resolution for achieving high spatiotemporal resolution in hybrid camera system

    Science.gov (United States)

    Kamimura, Kenji; Tsumura, Norimichi; Nakaguchi, Toshiya; Miyake, Yoichi

    2010-05-01

    Many super-resolution methods have been proposed to enhance the spatial resolution of images by using iteration and multiple input images. In a previous paper, we proposed the example-based super-resolution method to enhance an image through pixel-based texton substitution to reduce the computational cost. In this method, however, we only considered the enhancement of a texture image. In this study, we modified this texton substitution method for a hybrid camera to reduce the required bandwidth of a high-resolution video camera. We applied our algorithm to pairs of high- and low-spatiotemporal-resolution videos, which were synthesized to simulate a hybrid camera. The result showed that the fine detail of the low-resolution video can be reproduced compared with bicubic interpolation and the required bandwidth could be reduced to about 1/5 in a video camera. It was also shown that the peak signal-to-noise ratios (PSNRs) of the images improved by about 6 dB in a trained frame and by 1.0-1.5 dB in a test frame, as determined by comparison with the processed image using bicubic interpolation, and the average PSNRs were higher than those obtained by the well-known Freeman’s patch-based super-resolution method. Compared with that of the Freeman’s patch-based super-resolution method, the computational time of our method was reduced to almost 1/10.

  20. Super-Resolution Molecular and Functional Imaging of Nanoscale Architectures in Life and Materials Science

    KAUST Repository

    Habuchi, Satoshi

    2014-06-12

    Super-resolution (SR) fluorescence microscopy has been revolutionizing the way in which we investigate the structures, dynamics, and functions of a wide range of nanoscale systems. In this review, I describe the current state of various SR fluorescence microscopy techniques along with the latest developments of fluorophores and labeling for the SR microscopy. I discuss the applications of SR microscopy in the fields of life science and materials science with a special emphasis on quantitative molecular imaging and nanoscale functional imaging. These studies open new opportunities for unraveling the physical, chemical, and optical properties of a wide range of nanoscale architectures together with their nanostructures and will enable the development of new (bio-)nanotechnology.

  1. Video Super-Resolution via Bidirectional Recurrent Convolutional Networks.

    Science.gov (United States)

    Huang, Yan; Wang, Wei; Wang, Liang

    2018-04-01

    Super resolving a low-resolution video, namely video super-resolution (SR), is usually handled by either single-image SR or multi-frame SR. Single-Image SR deals with each video frame independently, and ignores intrinsic temporal dependency of video frames which actually plays a very important role in video SR. Multi-Frame SR generally extracts motion information, e.g., optical flow, to model the temporal dependency, but often shows high computational cost. Considering that recurrent neural networks (RNNs) can model long-term temporal dependency of video sequences well, we propose a fully convolutional RNN named bidirectional recurrent convolutional network for efficient multi-frame SR. Different from vanilla RNNs, 1) the commonly-used full feedforward and recurrent connections are replaced with weight-sharing convolutional connections. So they can greatly reduce the large number of network parameters and well model the temporal dependency in a finer level, i.e., patch-based rather than frame-based, and 2) connections from input layers at previous timesteps to the current hidden layer are added by 3D feedforward convolutions, which aim to capture discriminate spatio-temporal patterns for short-term fast-varying motions in local adjacent frames. Due to the cheap convolutional operations, our model has a low computational complexity and runs orders of magnitude faster than other multi-frame SR methods. With the powerful temporal dependency modeling, our model can super resolve videos with complex motions and achieve well performance.

  2. Field-portable pixel super-resolution colour microscope.

    Directory of Open Access Journals (Sweden)

    Alon Greenbaum

    Full Text Available Based on partially-coherent digital in-line holography, we report a field-portable microscope that can render lensfree colour images over a wide field-of-view of e.g., >20 mm(2. This computational holographic microscope weighs less than 145 grams with dimensions smaller than 17×6×5 cm, making it especially suitable for field settings and point-of-care use. In this lensfree imaging design, we merged a colorization algorithm with a source shifting based multi-height pixel super-resolution technique to mitigate 'rainbow' like colour artefacts that are typical in holographic imaging. This image processing scheme is based on transforming the colour components of an RGB image into YUV colour space, which separates colour information from brightness component of an image. The resolution of our super-resolution colour microscope was characterized using a USAF test chart to confirm sub-micron spatial resolution, even for reconstructions that employ multi-height phase recovery to handle dense and connected objects. To further demonstrate the performance of this colour microscope Papanicolaou (Pap smears were also successfully imaged. This field-portable and wide-field computational colour microscope could be useful for tele-medicine applications in resource poor settings.

  3. Super-resolved linear fluorescence localization microscopy using photostable fluorophores: A virtual microscopy study

    Science.gov (United States)

    Birk, Udo; Szczurek, Aleksander; Cremer, Christoph

    2017-12-01

    Current approaches to overcome the conventional limit of the resolution potential of light microscopy (of about 200 nm for visible light), often suffer from non-linear effects, which render the quantification of the image intensities in the reconstructions difficult, and also affect the quantification of the biological structure under investigation. As an attempt to face these difficulties, we discuss a particular method of localization microscopy which is based on photostable fluorescent dyes. The proposed method can potentially be implemented as a fast alternative for quantitative localization microscopy, circumventing the need for the acquisition of thousands of image frames and complex, highly dye-specific imaging buffers. Although the need for calibration remains in order to extract quantitative data (such as the number of emitters), multispectral approaches are largely facilitated due to the much less stringent requirements on imaging buffers. Furthermore, multispectral acquisitions can be readily obtained using commercial instrumentation such as e.g. the conventional confocal laser scanning microscope.

  4. Wavelet Filter Banks for Super-Resolution SAR Imaging

    Science.gov (United States)

    Sheybani, Ehsan O.; Deshpande, Manohar; Memarsadeghi, Nargess

    2011-01-01

    This paper discusses Innovative wavelet-based filter banks designed to enhance the analysis of super resolution Synthetic Aperture Radar (SAR) images using parametric spectral methods and signal classification algorithms, SAR finds applications In many of NASA's earth science fields such as deformation, ecosystem structure, and dynamics of Ice, snow and cold land processes, and surface water and ocean topography. Traditionally, standard methods such as Fast-Fourier Transform (FFT) and Inverse Fast-Fourier Transform (IFFT) have been used to extract Images from SAR radar data, Due to non-parametric features of these methods and their resolution limitations and observation time dependence, use of spectral estimation and signal pre- and post-processing techniques based on wavelets to process SAR radar data has been proposed. Multi-resolution wavelet transforms and advanced spectral estimation techniques have proven to offer efficient solutions to this problem.

  5. Single image super-resolution based on convolutional neural networks

    Science.gov (United States)

    Zou, Lamei; Luo, Ming; Yang, Weidong; Li, Peng; Jin, Liujia

    2018-03-01

    We present a deep learning method for single image super-resolution (SISR). The proposed approach learns end-to-end mapping between low-resolution (LR) images and high-resolution (HR) images. The mapping is represented as a deep convolutional neural network which inputs the LR image and outputs the HR image. Our network uses 5 convolution layers, which kernels size include 5×5, 3×3 and 1×1. In our proposed network, we use residual-learning and combine different sizes of convolution kernels at the same layer. The experiment results show that our proposed method performs better than the existing methods in reconstructing quality index and human visual effects on benchmarked images.

  6. Super-resolution in plenoptic cameras using FPGAs.

    Science.gov (United States)

    Pérez, Joel; Magdaleno, Eduardo; Pérez, Fernando; Rodríguez, Manuel; Hernández, David; Corrales, Jaime

    2014-05-16

    Plenoptic cameras are a new type of sensor that extend the possibilities of current commercial cameras allowing 3D refocusing or the capture of 3D depths. One of the limitations of plenoptic cameras is their limited spatial resolution. In this paper we describe a fast, specialized hardware implementation of a super-resolution algorithm for plenoptic cameras. The algorithm has been designed for field programmable graphic array (FPGA) devices using VHDL (very high speed integrated circuit (VHSIC) hardware description language). With this technology, we obtain an acceleration of several orders of magnitude using its extremely high-performance signal processing capability through parallelism and pipeline architecture. The system has been developed using generics of the VHDL language. This allows a very versatile and parameterizable system. The system user can easily modify parameters such as data width, number of microlenses of the plenoptic camera, their size and shape, and the super-resolution factor. The speed of the algorithm in FPGA has been successfully compared with the execution using a conventional computer for several image sizes and different 3D refocusing planes.

  7. Super-Resolution in Plenoptic Cameras Using FPGAs

    Directory of Open Access Journals (Sweden)

    Joel Pérez

    2014-05-01

    Full Text Available Plenoptic cameras are a new type of sensor that extend the possibilities of current commercial cameras allowing 3D refocusing or the capture of 3D depths. One of the limitations of plenoptic cameras is their limited spatial resolution. In this paper we describe a fast, specialized hardware implementation of a super-resolution algorithm for plenoptic cameras. The algorithm has been designed for field programmable graphic array (FPGA devices using VHDL (very high speed integrated circuit (VHSIC hardware description language. With this technology, we obtain an acceleration of several orders of magnitude using its extremely high-performance signal processing capability through parallelism and pipeline architecture. The system has been developed using generics of the VHDL language. This allows a very versatile and parameterizable system. The system user can easily modify parameters such as data width, number of microlenses of the plenoptic camera, their size and shape, and the super-resolution factor. The speed of the algorithm in FPGA has been successfully compared with the execution using a conventional computer for several image sizes and different 3D refocusing planes.

  8. Super-Resolution Algorithm in Cumulative Virtual Blanking

    Science.gov (United States)

    Montillet, J. P.; Meng, X.; Roberts, G. W.; Woolfson, M. S.

    2008-11-01

    The proliferation of mobile devices and the emergence of wireless location-based services have generated consumer demand for precise location. In this paper, the MUSIC super-resolution algorithm is applied to time delay estimation for positioning purposes in cellular networks. The goal is to position a Mobile Station with UMTS technology. The problem of Base-Stations herability is solved using Cumulative Virtual Blanking. A simple simulator is presented using DS-SS signal. The results show that MUSIC algorithm improves the time delay estimation in both the cases whether or not Cumulative Virtual Blanking was carried out.

  9. Super-resolution with a positive epsilon multi-quantum-well super-lens

    International Nuclear Information System (INIS)

    Bak, A. O.; Giannini, V.; Maier, S. A.; Phillips, C. C.

    2013-01-01

    We design an anisotropic and dichroic quantum metamaterial that is able to achieve super-resolution without the need for a negative permittivity. When exploring the parameters of the structure, we take into account the limits of semiconductor fabrication technology based on quantum well stacks. By heavily doping the structure with free electrons, we infer an anisotropic effective medium with a prolate ellipsoid dispersion curve which allows for near-diffractionless propagation of light (similar to an epsilon-near-zero hyperbolic lens). This, coupled with low absorption, allows us to resolve images at the sub-wavelength scale at distances 6 times greater than equivalent natural materials

  10. 3D high- and super-resolution imaging using single-objective SPIM.

    Science.gov (United States)

    Galland, Remi; Grenci, Gianluca; Aravind, Ajay; Viasnoff, Virgile; Studer, Vincent; Sibarita, Jean-Baptiste

    2015-07-01

    Single-objective selective-plane illumination microscopy (soSPIM) is achieved with micromirrored cavities combined with a laser beam-steering unit installed on a standard inverted microscope. The illumination and detection are done through the same objective. soSPIM can be used with standard sample preparations and features high background rejection and efficient photon collection, allowing for 3D single-molecule-based super-resolution imaging of whole cells or cell aggregates. Using larger mirrors enabled us to broaden the capabilities of our system to image Drosophila embryos.

  11. Improved methods for high resolution electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, J.R.

    1987-04-01

    Existing methods of making support films for high resolution transmission electron microscopy are investigated and novel methods are developed. Existing methods of fabricating fenestrated, metal reinforced specimen supports (microgrids) are evaluated for their potential to reduce beam induced movement of monolamellar crystals of C/sub 44/H/sub 90/ paraffin supported on thin carbon films. Improved methods of producing hydrophobic carbon films by vacuum evaporation, and improved methods of depositing well ordered monolamellar paraffin crystals on carbon films are developed. A novel technique for vacuum evaporation of metals is described which is used to reinforce microgrids. A technique is also developed to bond thin carbon films to microgrids with a polymer bonding agent. Unique biochemical methods are described to accomplish site specific covalent modification of membrane proteins. Protocols are given which covalently convert the carboxy terminus of papain cleaved bacteriorhodopsin to a free thiol. 53 refs., 19 figs., 1 tab.

  12. Color image guided depth image super resolution using fusion filter

    Science.gov (United States)

    He, Jin; Liang, Bin; He, Ying; Yang, Jun

    2018-04-01

    Depth cameras are currently playing an important role in many areas. However, most of them can only obtain lowresolution (LR) depth images. Color cameras can easily provide high-resolution (HR) color images. Using color image as a guide image is an efficient way to get a HR depth image. In this paper, we propose a depth image super resolution (SR) algorithm, which uses a HR color image as a guide image and a LR depth image as input. We use the fusion filter of guided filter and edge based joint bilateral filter to get HR depth image. Our experimental results on Middlebury 2005 datasets show that our method can provide better quality in HR depth images both numerically and visually.

  13. Optimized multiple linear mappings for single image super-resolution

    Science.gov (United States)

    Zhang, Kaibing; Li, Jie; Xiong, Zenggang; Liu, Xiuping; Gao, Xinbo

    2017-12-01

    Learning piecewise linear regression has been recognized as an effective way for example learning-based single image super-resolution (SR) in literature. In this paper, we employ an expectation-maximization (EM) algorithm to further improve the SR performance of our previous multiple linear mappings (MLM) based SR method. In the training stage, the proposed method starts with a set of linear regressors obtained by the MLM-based method, and then jointly optimizes the clustering results and the low- and high-resolution subdictionary pairs for regression functions by using the metric of the reconstruction errors. In the test stage, we select the optimal regressor for SR reconstruction by accumulating the reconstruction errors of m-nearest neighbors in the training set. Thorough experimental results carried on six publicly available datasets demonstrate that the proposed SR method can yield high-quality images with finer details and sharper edges in terms of both quantitative and perceptual image quality assessments.

  14. Hierarchical super-structure identified by polarized light microscopy, electron microscopy and nanoindentation: Implications for the limits of biological control over the growth mode of abalone sea shells

    Directory of Open Access Journals (Sweden)

    Schneider Andreas S

    2012-09-01

    Full Text Available Abstract Background Mollusc shells are commonly investigated using high-resolution imaging techniques based on cryo-fixation. Less detailed information is available regarding the light-optical properties. Sea shells of Haliotis pulcherina were embedded for polishing in defined orientations in order to investigate the interface between prismatic calcite and nacreous aragonite by standard materialographic methods. A polished thin section of the interface was prepared with a defined thickness of 60 μm for quantitative birefringence analysis using polarized light and LC-PolScope microscopy. Scanning electron microscopy images were obtained for comparison. In order to study structural-mechanical relationships, nanoindentation experiments were performed. Results Incident light microscopy revealed a super-structure in semi-transparent regions of the polished cross-section under a defined angle. This super-structure is not visible in transmitted birefringence analysis due to the blurred polarization of small nacre platelets and numerous organic interfaces. The relative orientation and homogeneity of calcite prisms was directly identified, some of them with their optical axes exactly normal to the imaging plane. Co-oriented "prism colonies" were identified by polarized light analyses. The nacreous super-structure was also visualized by secondary electron imaging under defined angles. The domains of the super-structure were interpreted to consist of crystallographically aligned platelet stacks. Nanoindentation experiments showed that mechanical properties changed with the same periodicity as the domain size. Conclusions In this study, we have demonstrated that insights into the growth mechanisms of nacre can be obtained by conventional light-optical methods. For example, we observed super-structures formed by co-oriented nacre platelets as previously identified using X-ray Photo-electron Emission Microscopy (X-PEEM [Gilbert et al., Journal of the

  15. Sparsity-Based Super Resolution for SEM Images.

    Science.gov (United States)

    Tsiper, Shahar; Dicker, Or; Kaizerman, Idan; Zohar, Zeev; Segev, Mordechai; Eldar, Yonina C

    2017-09-13

    The scanning electron microscope (SEM) is an electron microscope that produces an image of a sample by scanning it with a focused beam of electrons. The electrons interact with the atoms in the sample, which emit secondary electrons that contain information about the surface topography and composition. The sample is scanned by the electron beam point by point, until an image of the surface is formed. Since its invention in 1942, the capabilities of SEMs have become paramount in the discovery and understanding of the nanometer world, and today it is extensively used for both research and in industry. In principle, SEMs can achieve resolution better than one nanometer. However, for many applications, working at subnanometer resolution implies an exceedingly large number of scanning points. For exactly this reason, the SEM diagnostics of microelectronic chips is performed either at high resolution (HR) over a small area or at low resolution (LR) while capturing a larger portion of the chip. Here, we employ sparse coding and dictionary learning to algorithmically enhance low-resolution SEM images of microelectronic chips-up to the level of the HR images acquired by slow SEM scans, while considerably reducing the noise. Our methodology consists of two steps: an offline stage of learning a joint dictionary from a sequence of LR and HR images of the same region in the chip, followed by a fast-online super-resolution step where the resolution of a new LR image is enhanced. We provide several examples with typical chips used in the microelectronics industry, as well as a statistical study on arbitrary images with characteristic structural features. Conceptually, our method works well when the images have similar characteristics, as microelectronics chips do. This work demonstrates that employing sparsity concepts can greatly improve the performance of SEM, thereby considerably increasing the scanning throughput without compromising on analysis quality and resolution.

  16. Adaptive Markov Random Fields for Example-Based Super-resolution of Faces

    Science.gov (United States)

    Stephenson, Todd A.; Chen, Tsuhan

    2006-12-01

    Image enhancement of low-resolution images can be done through methods such as interpolation, super-resolution using multiple video frames, and example-based super-resolution. Example-based super-resolution, in particular, is suited to images that have a strong prior (for those frameworks that work on only a single image, it is more like image restoration than traditional, multiframe super-resolution). For example, hallucination and Markov random field (MRF) methods use examples drawn from the same domain as the image being enhanced to determine what the missing high-frequency information is likely to be. We propose to use even stronger prior information by extending MRF-based super-resolution to use adaptive observation and transition functions, that is, to make these functions region-dependent. We show with face images how we can adapt the modeling for each image patch so as to improve the resolution.

  17. Adaptive Markov Random Fields for Example-Based Super-resolution of Faces

    Directory of Open Access Journals (Sweden)

    Stephenson Todd A

    2006-01-01

    Full Text Available Image enhancement of low-resolution images can be done through methods such as interpolation, super-resolution using multiple video frames, and example-based super-resolution. Example-based super-resolution, in particular, is suited to images that have a strong prior (for those frameworks that work on only a single image, it is more like image restoration than traditional, multiframe super-resolution. For example, hallucination and Markov random field (MRF methods use examples drawn from the same domain as the image being enhanced to determine what the missing high-frequency information is likely to be. We propose to use even stronger prior information by extending MRF-based super-resolution to use adaptive observation and transition functions, that is, to make these functions region-dependent. We show with face images how we can adapt the modeling for each image patch so as to improve the resolution.

  18. Resolution Versus Error for Computational Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Luzi, Lorenzo; Stevens, Andrew; Yang, Hao; Browning, Nigel D.

    2017-07-01

    Images that are collected via scanning transmission electron microscopy (STEM) can be undersampled to avoid damage to the specimen while maintaining resolution [1, 2]. We have used BPFA to impute missing data and reduce noise [3]. The reconstruction is typically evaluated using the peak signal-to-noise ratio (PSNR). This measure is too conservative for STEM images and we propose that the Fourier ring correlation (FRC) is used instead to evaluate the reconstruction. We are not concerned with exact reconstruction of the truth image, and therefore PSNR is a conservative estimation of the quality of the reconstruction. Instead, we are concerned with the visual resolution of the image and whether atoms can be distinguished. We have evaluated the reconstruction of a simulated STEM image using the FRC and compared the results with the PSNR measurements. The FRC captures the resolution of the image and is not affected by a large MSE if the atom peaks are still distinguishable. The noisy and reconstructed images are shown in Figure 1. The simulated STEM image was sampled at 100%, 80%, 40%, and 20% of the original pixels to simulate an undersampled scan. The reconstruction was done using BPFA with a patch size of 10 x 10 and no overlapping patches. Not having overlapping patches produces inferior results but they are still acceptable. The dictionary size is 64 and 30 iterations were completed during each reconstruction. The 100% image was denoised instead of reconstructed. Poisson noise was applied to the simulated image with λ values of 500, 50, and 5 to simulate lower imaging dose. The original simulated STEM image was also included in our calculations and was generated using a dose of 1000. The simulated STEM image is 100 by 100 pixels and has essentially no high frequency components. The image reconstruction tends to smooth the data, also resulting in no high frequency components. This causes the FRC of the two images to be large at higher resolutions and may be

  19. Application of Super-Resolution Convolutional Neural Network for Enhancing Image Resolution in Chest CT.

    Science.gov (United States)

    Umehara, Kensuke; Ota, Junko; Ishida, Takayuki

    2017-10-18

    In this study, the super-resolution convolutional neural network (SRCNN) scheme, which is the emerging deep-learning-based super-resolution method for enhancing image resolution in chest CT images, was applied and evaluated using the post-processing approach. For evaluation, 89 chest CT cases were sampled from The Cancer Imaging Archive. The 89 CT cases were divided randomly into 45 training cases and 44 external test cases. The SRCNN was trained using the training dataset. With the trained SRCNN, a high-resolution image was reconstructed from a low-resolution image, which was down-sampled from an original test image. For quantitative evaluation, two image quality metrics were measured and compared to those of the conventional linear interpolation methods. The image restoration quality of the SRCNN scheme was significantly higher than that of the linear interpolation methods (p < 0.001 or p < 0.05). The high-resolution image reconstructed by the SRCNN scheme was highly restored and comparable to the original reference image, in particular, for a ×2 magnification. These results indicate that the SRCNN scheme significantly outperforms the linear interpolation methods for enhancing image resolution in chest CT images. The results also suggest that SRCNN may become a potential solution for generating high-resolution CT images from standard CT images.

  20. Performance Evaluations for Super-Resolution Mosaicing on UAS Surveillance Videos

    Directory of Open Access Journals (Sweden)

    Aldo Camargo

    2013-05-01

    Full Text Available Abstract Unmanned Aircraft Systems (UAS have been widely applied for reconnaissance and surveillance by exploiting information collected from the digital imaging payload. The super-resolution (SR mosaicing of low-resolution (LR UAS surveillance video frames has become a critical requirement for UAS video processing and is important for further effective image understanding. In this paper we develop a novel super-resolution framework, which does not require the construction of sparse matrices. The proposed method implements image operations in the spatial domain and applies an iterated back-projection to construct super-resolution mosaics from the overlapping UAS surveillance video frames. The Steepest Descent method, the Conjugate Gradient method and the Levenberg-Marquardt algorithm are used to numerically solve the nonlinear optimization problem for estimating a super-resolution mosaic. A quantitative performance comparison in terms of computation time and visual quality of the super-resolution mosaics through the three numerical techniques is presented.

  1. Learning-based compressed sensing for infrared image super resolution

    Science.gov (United States)

    Zhao, Yao; Sui, Xiubao; Chen, Qian; Wu, Shaochi

    2016-05-01

    This paper presents an infrared image super-resolution method based on compressed sensing (CS). First, the reconstruction model under the CS framework is established and a Toeplitz matrix is selected as the sensing matrix. Compared with traditional learning-based methods, the proposed method uses a set of sub-dictionaries instead of two coupled dictionaries to recover high resolution (HR) images. And Toeplitz sensing matrix allows the proposed method time-efficient. Second, all training samples are divided into several feature spaces by using the proposed adaptive k-means classification method, which is more accurate than the standard k-means method. On the basis of this approach, a complex nonlinear mapping from the HR space to low resolution (LR) space can be converted into several compact linear mappings. Finally, the relationships between HR and LR image patches can be obtained by multi-sub-dictionaries and HR infrared images are reconstructed by the input LR images and multi-sub-dictionaries. The experimental results show that the proposed method is quantitatively and qualitatively more effective than other state-of-the-art methods.

  2. Super-resolution for a point source using positive refraction

    Science.gov (United States)

    Miñano, Juan C.; Benítez, Pablo; González, Juan C.; Grabovičkić, Dejan; Ahmadpanahi, Hamed

    Leonhardt demonstrated (2009) that the 2D Maxwell Fish Eye lens (MFE) can focus perfectly 2D Helmholtz waves of arbitrary frequency, i.e., it can transport perfectly an outward (monopole) 2D Helmholtz wave field, generated by a point source, towards a receptor called "perfect drain" (PD) located at the corresponding MFE image point. The PD has the property of absorbing the complete radiation without radiation or scattering and it has been claimed as necessary to obtain super-resolution (SR) in the MFE. However, a prototype using a "drain" different from the PD has shown λ/5 resolution for microwave frequencies (Ma et al, 2010). Recently, the SR properties of a device equivalent to the MFE, called the Spherical Geodesic Waveguide (SGW) (Miñano et al, 2012) have been analyzed. The reported results show resolution up to λ /3000, for the SGW loaded with the perfect drain, and up to λ /500 for the SGW without perfect drain. The perfect drain was realized as a coaxial probe loaded with properly calculated impedance. The SGW provides SR only in a narrow band of frequencies close to the resonance Schumann frequencies. Here we analyze the SGW loaded with a small "perfect drain region" (González et al, 2011). This drain is designed as a region made of a material with complex permittivity. The comparative results show that there is no significant difference in the SR properties for both perfect drain designs.

  3. Zeolites - a high resolution electron microscopy study

    International Nuclear Information System (INIS)

    Alfredsson, V.

    1994-10-01

    High resolution transmission electron microscopy (HRTEM) has been used to investigate a number of zeolites (EMT, FAU, LTL, MFI and MOR) and a member of the mesoporous M41S family. The electron optical artefact, manifested as a dark spot in the projected centre of the large zeolite channels, caused by insufficient transfer of certain reflections in the objective lens has been explained. The artefact severely hinders observation of materials confined in the zeolite channels and cavities. It is shown how to circumvent the artefact problem and how to image confined materials in spite of disturbance caused by the artefact. Image processing by means of a Wiener filter has been applied for removal of the artefact. The detailed surface structure of FAU has been investigated. Comparison of experimental micrographs with images simulated using different surface models indicates that the surface can be terminated in different ways depending on synthesis methods. The dealuminated form of FAU (USY) is covered by an amorphous region. Platinum incorporated in FAU has a preponderance to aggregate in the (111) twin planes, probably due to a local difference in cage structure with more spacious cages. It is shown that platinum is intra-zeolitic as opposed to being located on the external surface of the zeolite crystal. This could be deduced from tomography of ultra-thin sections among observations. HRTEM studies of the mesoporous MCM-41 show that the pores have a hexagonal shape and also supports the mechanistic model proposed which involves a cooperative formation of a mesophase including the silicate species as well as the surfactant. 66 refs, 24 figs

  4. Super Resolution Fluorescence Microscopy and Tracking of Bacterial Flotillin (Reggie Paralogs Provide Evidence for Defined-Sized Protein Microdomains within the Bacterial Membrane but Absence of Clusters Containing Detergent-Resistant Proteins.

    Directory of Open Access Journals (Sweden)

    Felix Dempwolff

    2016-06-01

    Full Text Available Biological membranes have been proposed to contain microdomains of a specific lipid composition, in which distinct groups of proteins are clustered. Flotillin-like proteins are conserved between pro-and eukaryotes, play an important function in several eukaryotic and bacterial cells, and define in vertebrates a type of so-called detergent-resistant microdomains. Using STED microscopy, we show that two bacterial flotillins, FloA and FloT, form defined assemblies with an average diameter of 85 to 110 nm in the model bacterium Bacillus subtilis. Interestingly, flotillin microdomains are of similar size in eukaryotic cells. The soluble domains of FloA form higher order oligomers of up to several hundred kDa in vitro, showing that like eukaryotic flotillins, bacterial assemblies are based in part on their ability to self-oligomerize. However, B. subtilis paralogs show significantly different diffusion rates, and consequently do not colocalize into a common microdomain. Dual colour time lapse experiments of flotillins together with other detergent-resistant proteins in bacteria show that proteins colocalize for no longer than a few hundred milliseconds, and do not move together. Our data reveal that the bacterial membrane contains defined-sized protein domains rather than functional microdomains dependent on flotillins. Based on their distinct dynamics, FloA and FloT confer spatially distinguishable activities, but do not serve as molecular scaffolds.

  5. A Super-resolution Reconstruction Algorithm for Surveillance Video

    Directory of Open Access Journals (Sweden)

    Jian Shao

    2017-01-01

    Full Text Available Recent technological developments have resulted in surveillance video becoming a primary method of preserving public security. Many city crimes are observed in surveillance video. The most abundant evidence collected by the police is also acquired through surveillance video sources. Surveillance video footage offers very strong support for solving criminal cases, therefore, creating an effective policy, and applying useful methods to the retrieval of additional evidence is becoming increasingly important. However, surveillance video has had its failings, namely, video footage being captured in low resolution (LR and bad visual quality. In this paper, we discuss the characteristics of surveillance video and describe the manual feature registration – maximum a posteriori – projection onto convex sets to develop a super-resolution reconstruction method, which improves the quality of surveillance video. From this method, we can make optimal use of information contained in the LR video image, but we can also control the image edge clearly as well as the convergence of the algorithm. Finally, we make a suggestion on how to adjust the algorithm adaptability by analyzing the prior information of target image.

  6. Super-resolution for imagery from integrated microgrid polarimeters.

    Science.gov (United States)

    Hardie, Russell C; LeMaster, Daniel A; Ratliff, Bradley M

    2011-07-04

    Imagery from microgrid polarimeters is obtained by using a mosaic of pixel-wise micropolarizers on a focal plane array (FPA). Each distinct polarization image is obtained by subsampling the full FPA image. Thus, the effective pixel pitch for each polarization channel is increased and the sampling frequency is decreased. As a result, aliasing artifacts from such undersampling can corrupt the true polarization content of the scene. Here we present the first multi-channel multi-frame super-resolution (SR) algorithms designed specifically for the problem of image restoration in microgrid polarization imagers. These SR algorithms can be used to address aliasing and other degradations, without sacrificing field of view or compromising optical resolution with an anti-aliasing filter. The new SR methods are designed to exploit correlation between the polarimetric channels. One of the new SR algorithms uses a form of regularized least squares and has an iterative solution. The other is based on the faster adaptive Wiener filter SR method. We demonstrate that the new multi-channel SR algorithms are capable of providing significant enhancement of polarimetric imagery and that they outperform their independent channel counterparts.

  7. Image quality assessment for selfies with and without super resolution

    Science.gov (United States)

    Kubota, Aya; Gohshi, Seiichi

    2018-04-01

    With the advent of cellphone cameras, in particular, on smartphones, many people now take photos of themselves alone and with others in the frame; such photos are popularly known as "selfies". Most smartphones are equipped with two cameras: the front-facing and rear cameras. The camera located on the back of the smartphone is referred to as the "out-camera," whereas the one located on the front of the smartphone is called the "in-camera." In-cameras are mainly used for selfies. Some smartphones feature high-resolution cameras. However, the original image quality cannot be obtained because smartphone cameras often have low-performance lenses. Super resolution (SR) is one of the recent technological advancements that has increased image resolution. We developed a new SR technology that can be processed on smartphones. Smartphones with new SR technology are currently available in the market have already registered sales. However, the effective use of new SR technology has not yet been verified. Comparing the image quality with and without SR on smartphone display is necessary to confirm the usefulness of this new technology. Methods that are based on objective and subjective assessments are required to quantitatively measure image quality. It is known that the typical object assessment value, such as Peak Signal to Noise Ratio (PSNR), does not go together with how we feel when we assess image/video. When digital broadcast started, the standard was determined using subjective assessment. Although subjective assessment usually comes at high cost because of personnel expenses for observers, the results are highly reproducible when they are conducted under right conditions and statistical analysis. In this study, the subjective assessment results for selfie images are reported.

  8. Atomic resolution three-dimensional electron diffraction microscopy

    International Nuclear Information System (INIS)

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

    2002-01-01

    We report the development of a novel form of diffraction-based 3D microscopy to overcome resolution barriers inherent in high-resolution electron microscopy and tomography. By combining coherent electron diffraction with the oversampling phasing method, we show that the 3D structure of a nanocrystal can be determined ab initio at a resolution of 1 Angstrom from 29 simulated noisy diffraction patterns. This new form of microscopy can be used to image the 3D structures of nanocrystals and noncrystalline samples, with resolution limited only by the quality of sample diffraction

  9. Super-resolution with an optically-addressable liquid crystal spatial light modulator

    International Nuclear Information System (INIS)

    McOrist, J.; Sharma, M.D.; Sheppard, C.J.R.

    2002-01-01

    Full text: An optically-addressable liquid crystal spatial light modulator has been used to generate super-resolving masks. This approach avoids problems of low efficiency and coupling between amplitude and phase modulation, that occur when using conventional liquid crystal modulators. When addressed by a programmed light intensity distribution, it allows filters to be changed rapidly to modify the response of a system or permit the investigation of different filter designs. The device used is not pixellated, with a spatial resolution of 30 line pairs/mm over an area 18mm X 18mm, and can achieve continuously-variable phase modulation up to 1.5 wavelengths. The system consists of a write-beam that is collimated from a white-light source. An input mask was used in our experiments determines the modulation pattern of the read-beam. The read-beam from a HeNe laser reflects from the modulator and is focused by a microscope objective. The value of the phase change induced by the transparent regions of the mask can be altered continuously by adjusting the brightness of the write-beam. We have used this system to attain super-resolution by simple Toraldo filters, consisting of arrays of rings. Copyright (2002) Australian Society for Electron Microscopy Inc

  10. Resolution enhancement of tri-stereo remote sensing images by super resolution methods

    Science.gov (United States)

    Tuna, Caglayan; Akoguz, Alper; Unal, Gozde; Sertel, Elif

    2016-10-01

    Super resolution (SR) refers to generation of a High Resolution (HR) image from a decimated, blurred, low-resolution (LR) image set, which can be either a single frame or multi-frame that contains a collection of several images acquired from slightly different views of the same observation area. In this study, we propose a novel application of tri-stereo Remote Sensing (RS) satellite images to the super resolution problem. Since the tri-stereo RS images of the same observation area are acquired from three different viewing angles along the flight path of the satellite, these RS images are properly suited to a SR application. We first estimate registration between the chosen reference LR image and other LR images to calculate the sub pixel shifts among the LR images. Then, the warping, blurring and down sampling matrix operators are created as sparse matrices to avoid high memory and computational requirements, which would otherwise make the RS-SR solution impractical. Finally, the overall system matrix, which is constructed based on the obtained operator matrices is used to obtain the estimate HR image in one step in each iteration of the SR algorithm. Both the Laplacian and total variation regularizers are incorporated separately into our algorithm and the results are presented to demonstrate an improved quantitative performance against the standard interpolation method as well as improved qualitative results due expert evaluations.

  11. Overcoming Registration Uncertainty in Image Super-Resolution: Maximize or Marginalize?

    Directory of Open Access Journals (Sweden)

    Andrew Zisserman

    2007-01-01

    Full Text Available In multiple-image super-resolution, a high-resolution image is estimated from a number of lower-resolution images. This usually involves computing the parameters of a generative imaging model (such as geometric and photometric registration, and blur and obtaining a MAP estimate by minimizing a cost function including an appropriate prior. Two alternative approaches are examined. First, both registrations and the super-resolution image are found simultaneously using a joint MAP optimization. Second, we perform Bayesian integration over the unknown image registration parameters, deriving a cost function whose only variables of interest are the pixel values of the super-resolution image. We also introduce a scheme to learn the parameters of the image prior as part of the super-resolution algorithm. We show examples on a number of real sequences including multiple stills, digital video, and DVDs of movies.

  12. Quantitative transmission electron microscopy at atomic resolution

    International Nuclear Information System (INIS)

    Allen, L J; D'Alfonso, A J; Forbes, B D; Findlay, S D; LeBeau, J M; Stemmer, S

    2012-01-01

    In scanning transmission electron microscopy (STEM) it is possible to operate the microscope in bright-field mode under conditions which, by the quantum mechanical principle of reciprocity, are equivalent to those in conventional transmission electron microscopy (CTEM). The results of such an experiment will be presented which are in excellent quantitative agreement with theory for specimens up to 25 nm thick. This is at variance with the large contrast mismatch (typically between two and five) noted in equivalent CTEM experiments. The implications of this will be discussed.

  13. The tetraspanin web revisited by super-resolution microscopy

    NARCIS (Netherlands)

    Zuidscherwoude, M.C.M.; Gottfert, F.; Dunlock, V.M.; Figdor, C.G.; Bogaart, G. van den; Spriel, A.B. van

    2015-01-01

    The spatial organization of membrane proteins in the plasma membrane is critical for signal transduction, cell communication and membrane trafficking. Tetraspanins organize functional higher-order protein complexes called 'tetraspanin-enriched microdomains (TEMs)' via interactions with partner

  14. Super-resolution imaging and tracking of protein-protein interactions in sub-diffraction cellular space

    Science.gov (United States)

    Liu, Zhen; Xing, Dong; Su, Qian Peter; Zhu, Yun; Zhang, Jiamei; Kong, Xinyu; Xue, Boxin; Wang, Sheng; Sun, Hao; Tao, Yile; Sun, Yujie

    2014-07-01

    Imaging the location and dynamics of individual interacting protein pairs is essential but often difficult because of the fluorescent background from other paired and non-paired molecules, particularly in the sub-diffraction cellular space. Here we develop a new method combining bimolecular fluorescence complementation and photoactivated localization microscopy for super-resolution imaging and single-molecule tracking of specific protein-protein interactions. The method is used to study the interaction of two abundant proteins, MreB and EF-Tu, in Escherichia coli cells. The super-resolution imaging shows interesting distribution and domain sizes of interacting MreB-EF-Tu pairs as a subpopulation of total EF-Tu. The single-molecule tracking of MreB, EF-Tu and MreB-EF-Tu pairs reveals intriguing localization-dependent heterogonous dynamics and provides valuable insights to understanding the roles of MreB-EF-Tu interactions.

  15. Super-resolution imaging and tracking of protein–protein interactions in sub-diffraction cellular space

    Science.gov (United States)

    Liu, Zhen; Xing, Dong; Su, Qian Peter; Zhu, Yun; Zhang, Jiamei; Kong, Xinyu; Xue, Boxin; Wang, Sheng; Sun, Hao; Tao, Yile; Sun, Yujie

    2014-01-01

    Imaging the location and dynamics of individual interacting protein pairs is essential but often difficult because of the fluorescent background from other paired and non-paired molecules, particularly in the sub-diffraction cellular space. Here we develop a new method combining bimolecular fluorescence complementation and photoactivated localization microscopy for super-resolution imaging and single-molecule tracking of specific protein–protein interactions. The method is used to study the interaction of two abundant proteins, MreB and EF-Tu, in Escherichia coli cells. The super-resolution imaging shows interesting distribution and domain sizes of interacting MreB–EF-Tu pairs as a subpopulation of total EF-Tu. The single-molecule tracking of MreB, EF-Tu and MreB–EF-Tu pairs reveals intriguing localization-dependent heterogonous dynamics and provides valuable insights to understanding the roles of MreB–EF-Tu interactions. PMID:25030837

  16. Magnetic Resonance Super-resolution Imaging Measurement with Dictionary-optimized Sparse Learning

    Directory of Open Access Journals (Sweden)

    Li Jun-Bao

    2017-06-01

    Full Text Available Magnetic Resonance Super-resolution Imaging Measurement (MRIM is an effective way of measuring materials. MRIM has wide applications in physics, chemistry, biology, geology, medical and material science, especially in medical diagnosis. It is feasible to improve the resolution of MR imaging through increasing radiation intensity, but the high radiation intensity and the longtime of magnetic field harm the human body. Thus, in the practical applications the resolution of hardware imaging reaches the limitation of resolution. Software-based super-resolution technology is effective to improve the resolution of image. This work proposes a framework of dictionary-optimized sparse learning based MR super-resolution method. The framework is to solve the problem of sample selection for dictionary learning of sparse reconstruction. The textural complexity-based image quality representation is proposed to choose the optimal samples for dictionary learning. Comprehensive experiments show that the dictionary-optimized sparse learning improves the performance of sparse representation.

  17. High-resolution and super stacking of time-reversal mirrors in locating seismic sources

    KAUST Repository

    Cao, Weiping; Hanafy, Sherif M.; Schuster, Gerard T.; Zhan, Ge; Boonyasiriwat, Chaiwoot

    2011-01-01

    Time reversal mirrors can be used to backpropagate and refocus incident wavefields to their actual source location, with the subsequent benefits of imaging with high-resolution and super-stacking properties. These benefits of time reversal mirrors

  18. Super-resolution imaging with Pontamine Fast Scarlet 4BS enables direct visualization of cellulose orientation and cell connection architecture in onion epidermis cells

    DEFF Research Database (Denmark)

    Liesche, Johannes; Ziomkiewicz, Iwona; Schulz, Alexander

    2013-01-01

    of cellulose fibril orientation and growth. The fluorescent dye Pontamine Fast Scarlet 4BS (PFS) was shown to stain cellulose with high specificity and could be used to visualize cellulose bundles in cell walls of Arabidopsis root epidermal cells with confocal microscopy. The resolution limit of confocal...... present the first super-resolution images of cellulose bundles in the plant cell wall produced by direct stochastic optical reconstruction microscopy (dSTORM) in combination with total internal reflection fluorescence (TIRF) microscopy. Since TIRF limits observation to the cell surface, we tested...... as alternatives 3D-structured illumination microscopy (3D-SIM) and confocal microscopy, combined with image deconvolution. Both methods offer lower resolution than STORM, but enable 3D imaging. While 3D-SIM produced strong artifacts, deconvolution gave good results. The resolution was improved over conventional...

  19. Evaluation of deep neural networks for single image super-resolution in a maritime context

    NARCIS (Netherlands)

    Nieuwenhuizen, R.P.J.; Kruithof, M.; Schutte, K.

    2017-01-01

    High resolution imagery is of crucial importance for the performance on visual recognition tasks. Super-resolution (SR) reconstruction algorithms aim to enhance the image resolution beyond the capability of the image sensor being used. Traditional SR algorithms approach this inverse problem using

  20. Signal Characteristics of Super-Resolution Near-Field Structure Disks with 100 GB Capacity

    Science.gov (United States)

    Kim, Jooho; Hwang, Inoh; Kim, Hyunki; Park, Insik; Tominaga, Junji

    2005-05-01

    We report the basic characteristics of super resolution near-field structure (Super-RENS) media at a blue laser optical system (laser wavelength 405 nm, numerical aperture 0.85). Using a novel write once read many (WORM) structure for a blue laser system, we obtained a carrier-to-noise ratio (CNR) above 33 dB from the signal of the 37.5 nm mark length, which is equivalent to a 100 GB capacity with a 0.32 micrometer track pitch, and an eye pattern for 50 GB (2T: 75 nm) capacity using a patterned signal. Using a novel super-resolution material (tellurium, Te) with low super-resolution readout power, we also improved the read stability.

  1. Coherent optical adaptive technique improves the spatial resolution of STED microscopy in thick samples

    Science.gov (United States)

    Yan, Wei; Yang, Yanlong; Tan, Yu; Chen, Xun; Li, Yang; Qu, Junle; Ye, Tong

    2018-01-01

    Stimulated emission depletion microscopy (STED) is one of far-field optical microscopy techniques that can provide sub-diffraction spatial resolution. The spatial resolution of the STED microscopy is determined by the specially engineered beam profile of the depletion beam and its power. However, the beam profile of the depletion beam may be distorted due to aberrations of optical systems and inhomogeneity of specimens’ optical properties, resulting in a compromised spatial resolution. The situation gets deteriorated when thick samples are imaged. In the worst case, the sever distortion of the depletion beam profile may cause complete loss of the super resolution effect no matter how much depletion power is applied to specimens. Previously several adaptive optics approaches have been explored to compensate aberrations of systems and specimens. However, it is hard to correct the complicated high-order optical aberrations of specimens. In this report, we demonstrate that the complicated distorted wavefront from a thick phantom sample can be measured by using the coherent optical adaptive technique (COAT). The full correction can effectively maintain and improve the spatial resolution in imaging thick samples. PMID:29400356

  2. Super-Resolution Enhancement From Multiple Overlapping Images: A Fractional Area Technique

    Science.gov (United States)

    Michaels, Joshua A.

    With the availability of large quantities of relatively low-resolution data from several decades of space borne imaging, methods of creating an accurate, higher-resolution image from the multiple lower-resolution images (i.e. super-resolution), have been developed almost since such imagery has been around. The fractional-area super-resolution technique developed in this thesis has never before been documented. Satellite orbits, like Landsat, have a quantifiable variation, which means each image is not centered on the exact same spot more than once and the overlapping information from these multiple images may be used for super-resolution enhancement. By splitting a single initial pixel into many smaller, desired pixels, a relationship can be created between them using the ratio of the area within the initial pixel. The ideal goal for this technique is to obtain smaller pixels with exact values and no error, yielding a better potential result than those methods that yield interpolated pixel values with consequential loss of spatial resolution. A Fortran 95 program was developed to perform all calculations associated with the fractional-area super-resolution technique. The fractional areas are calculated using traditional trigonometry and coordinate geometry and Linear Algebra Package (LAPACK; Anderson et al., 1999) is used to solve for the higher-resolution pixel values. In order to demonstrate proof-of-concept, a synthetic dataset was created using the intrinsic Fortran random number generator and Adobe Illustrator CS4 (for geometry). To test the real-life application, digital pictures from a Sony DSC-S600 digital point-and-shoot camera with a tripod were taken of a large US geological map under fluorescent lighting. While the fractional-area super-resolution technique works in perfect synthetic conditions, it did not successfully produce a reasonable or consistent solution in the digital photograph enhancement test. The prohibitive amount of processing time (up to

  3. Introduction to the virtual special issue on super-resolution imaging techniques

    Science.gov (United States)

    Cao, Liangcai; Liu, Zhengjun

    2017-12-01

    Until quite recently, the resolution of optical imaging instruments, including telescopes, cameras and microscopes, was considered to be limited by the diffraction of light and by image sensors. In the past few years, many exciting super-resolution approaches have emerged that demonstrate intriguing ways to bypass the classical limit in optics and detectors. More and more research groups are engaged in the study of advanced super-resolution schemes, devices, algorithms, systems, and applications [1-6]. Super-resolution techniques involve new methods in science and engineering of optics [7,8], measurements [9,10], chemistry [11,12] and information [13,14]. Promising applications, particularly in biomedical research and semiconductor industry, have been successfully demonstrated.

  4. Low-Cost Super-Resolution Algorithms Implementation Over a HW/SW Video Compression Platform

    Directory of Open Access Journals (Sweden)

    Llopis Rafael Peset

    2006-01-01

    Full Text Available Two approaches are presented in this paper to improve the quality of digital images over the sensor resolution using super-resolution techniques: iterative super-resolution (ISR and noniterative super-resolution (NISR algorithms. The results show important improvements in the image quality, assuming that sufficient sample data and a reasonable amount of aliasing are available at the input images. These super-resolution algorithms have been implemented over a codesign video compression platform developed by Philips Research, performing minimal changes on the overall hardware architecture. In this way, a novel and feasible low-cost implementation has been obtained by using the resources encountered in a generic hybrid video encoder. Although a specific video codec platform has been used, the methodology presented in this paper is easily extendable to any other video encoder architectures. Finally a comparison in terms of memory, computational load, and image quality for both algorithms, as well as some general statements about the final impact of the sampling process on the quality of the super-resolved (SR image, are also presented.

  5. A graphene oxide-carbon nanotube grid for high-resolution transmission electron microscopy of nanomaterials

    International Nuclear Information System (INIS)

    Zhang Lina; Zhang Haoxu; Zhou Ruifeng; Chen Zhuo; Li Qunqing; Fan Shoushan; Jiang Kaili; Ge Guanglu; Liu Renxiao

    2011-01-01

    A novel grid for use in transmission electron microscopy is developed. The supporting film of the grid is composed of thin graphene oxide films overlying a super-aligned carbon nanotube network. The composite film combines the advantages of graphene oxide and carbon nanotube networks and has the following properties: it is ultra-thin, it has a large flat and smooth effective supporting area with a homogeneous amorphous appearance, high stability, and good conductivity. The graphene oxide-carbon nanotube grid has a distinct advantage when characterizing the fine structure of a mass of nanomaterials over conventional amorphous carbon grids. Clear high-resolution transmission electron microscopy images of various nanomaterials are obtained easily using the new grids.

  6. Fluorescence microscopy.

    Science.gov (United States)

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

    2014-10-01

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  7. Single Image Super-Resolution Based on Multi-Scale Competitive Convolutional Neural Network.

    Science.gov (United States)

    Du, Xiaofeng; Qu, Xiaobo; He, Yifan; Guo, Di

    2018-03-06

    Deep convolutional neural networks (CNNs) are successful in single-image super-resolution. Traditional CNNs are limited to exploit multi-scale contextual information for image reconstruction due to the fixed convolutional kernel in their building modules. To restore various scales of image details, we enhance the multi-scale inference capability of CNNs by introducing competition among multi-scale convolutional filters, and build up a shallow network under limited computational resources. The proposed network has the following two advantages: (1) the multi-scale convolutional kernel provides the multi-context for image super-resolution, and (2) the maximum competitive strategy adaptively chooses the optimal scale of information for image reconstruction. Our experimental results on image super-resolution show that the performance of the proposed network outperforms the state-of-the-art methods.

  8. Multiband super-resolution imaging of graded-index photonic crystal flat lens

    Science.gov (United States)

    Xie, Jianlan; Wang, Junzhong; Ge, Rui; Yan, Bei; Liu, Exian; Tan, Wei; Liu, Jianjun

    2018-05-01

    Multiband super-resolution imaging of point source is achieved by a graded-index photonic crystal flat lens. With the calculations of six bands in common photonic crystal (CPC) constructed with scatterers of different refractive indices, it can be found that the super-resolution imaging of point source can be realized by different physical mechanisms in three different bands. In the first band, the imaging of point source is based on far-field condition of spherical wave while in the second band, it is based on the negative effective refractive index and exhibiting higher imaging quality than that of the CPC. However, in the fifth band, the imaging of point source is mainly based on negative refraction of anisotropic equi-frequency surfaces. The novel method of employing different physical mechanisms to achieve multiband super-resolution imaging of point source is highly meaningful for the field of imaging.

  9. Ultrafast terahertz scanning tunneling microscopy with atomic resolution

    DEFF Research Database (Denmark)

    Jelic, Vedran; Iwaszczuk, Krzysztof; Nguyen, Peter H.

    2016-01-01

    We demonstrate that ultrafast terahertz scanning tunneling microscopy (THz-STM) can probe single atoms on a silicon surface with simultaneous sub-nanometer and sub-picosecond spatio-temporal resolution. THz-STM is established as a new technique for exploring high-field non-equilibrium tunneling...

  10. Fast resolution change in neutral helium atom microscopy

    Science.gov (United States)

    Flatabø, R.; Eder, S. D.; Ravn, A. K.; Samelin, B.; Greve, M. M.; Reisinger, T.; Holst, B.

    2018-05-01

    In neutral helium atom microscopy, a beam of atoms is scanned across a surface. Though still in its infancy, neutral helium microscopy has seen a rapid development over the last few years. The inertness and low energy of the helium atoms (less than 0.1 eV) combined with a very large depth of field and the fact that the helium atoms do not penetrate any solid material at low energies open the possibility for a non-destructive instrument that can measure topology on the nanoscale even on fragile and insulating surfaces. The resolution is determined by the beam spot size on the sample. Fast resolution change is an attractive property of a microscope because it allows different aspects of a sample to be investigated and makes it easier to identify specific features. However up till now it has not been possible to change the resolution of a helium microscope without breaking the vacuum and changing parts of the atom source. Here we present a modified source design, which allows fast, step wise resolution change. The basic design idea is to insert a moveable holder with a series of collimating apertures in front of the source, thus changing the effective source size of the beam and thereby the spot size on the surface and thus the microscope resolution. We demonstrate a design with 3 resolution steps. The number of resolution steps can easily be extended.

  11. PALM and STORM: unlocking live-cell super-resolution

    CSIR Research Space (South Africa)

    Henriques, R

    2011-05-01

    Full Text Available microscopy methods such as PALM and STORM to achieve this important goal. ? 2011 Wiley Periodicals, Inc. Biopolymers 95: 322?331, 2011. http://onlinelibrary.wiley.com/doi/10.1002/bip.21586/abstract ...

  12. Resolution enhancement in integral microscopy by physical interpolation.

    Science.gov (United States)

    Llavador, Anabel; Sánchez-Ortiga, Emilio; Barreiro, Juan Carlos; Saavedra, Genaro; Martínez-Corral, Manuel

    2015-08-01

    Integral-imaging technology has demonstrated its capability for computing depth images from the microimages recorded after a single shot. This capability has been shown in macroscopic imaging and also in microscopy. Despite the possibility of refocusing different planes from one snap-shot is crucial for the study of some biological processes, the main drawback in integral imaging is the substantial reduction of the spatial resolution. In this contribution we report a technique, which permits to increase the two-dimensional spatial resolution of the computed depth images in integral microscopy by a factor of √2. This is made by a double-shot approach, carried out by means of a rotating glass plate, which shifts the microimages in the sensor plane. We experimentally validate the resolution enhancement as well as we show the benefit of applying the technique to biological specimens.

  13. Correction of a Depth-Dependent Lateral Distortion in 3D Super-Resolution Imaging.

    Directory of Open Access Journals (Sweden)

    Lina Carlini

    Full Text Available Three-dimensional (3D localization-based super-resolution microscopy (SR requires correction of aberrations to accurately represent 3D structure. Here we show how a depth-dependent lateral shift in the apparent position of a fluorescent point source, which we term `wobble`, results in warped 3D SR images and provide a software tool to correct this distortion. This system-specific, lateral shift is typically > 80 nm across an axial range of ~ 1 μm. A theoretical analysis based on phase retrieval data from our microscope suggests that the wobble is caused by non-rotationally symmetric phase and amplitude aberrations in the microscope's pupil function. We then apply our correction to the bacterial cytoskeletal protein FtsZ in live bacteria and demonstrate that the corrected data more accurately represent the true shape of this vertically-oriented ring-like structure. We also include this correction method in a registration procedure for dual-color, 3D SR data and show that it improves target registration error (TRE at the axial limits over an imaging depth of 1 μm, yielding TRE values of < 20 nm. This work highlights the importance of correcting aberrations in 3D SR to achieve high fidelity between the measurements and the sample.

  14. Structural analysis of herpes simplex virus by optical super-resolution imaging

    Science.gov (United States)

    Laine, Romain F.; Albecka, Anna; van de Linde, Sebastian; Rees, Eric J.; Crump, Colin M.; Kaminski, Clemens F.

    2015-01-01

    Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.

  15. Super-resolution mapping of scaffold nucleoporins in the nuclear pore complex.

    Science.gov (United States)

    Ma, Jiong; Kelich, Joseph M; Junod, Samuel L; Yang, Weidong

    2017-04-01

    The nuclear pore complex (NPC), composed of ∼30 different nucleoporins (Nups), is one of the largest supramolecular structures in eukaryotic cells. Its octagonal ring scaffold perforates the nuclear envelope and features a unique molecular machinery that regulates nucleocytoplasmic transport. However, the precise copy number and the spatial location of each Nup in the native NPC remain obscure due to the inherent difficulty of counting and localizing proteins inside of the sub-micrometer supramolecular complex. Here, we combined super-resolution single-point edge-excitation subdiffraction (SPEED) microscopy and nanobody-specific labeling to reveal the spatial distribution of scaffold Nups within three separate layers in the native NPC with a precision of ∼3 nm. Our data reveal both the radial and axial spatial distributions for Pom121, Nup37 and Nup35 and provide evidence for their copy numbers of 8, 32 and 16, respectively, per NPC. This approach can help pave the path for mapping the entirety of Nups in native NPCs and also other structural components of macromolecular complexes. © 2017. Published by The Company of Biologists Ltd.

  16. Multi-example feature-constrained back-projection method for image super-resolution

    Institute of Scientific and Technical Information of China (English)

    Junlei Zhang; Dianguang Gai; Xin Zhang; Xuemei Li

    2017-01-01

    Example-based super-resolution algorithms,which predict unknown high-resolution image information using a relationship model learnt from known high- and low-resolution image pairs, have attracted considerable interest in the field of image processing. In this paper, we propose a multi-example feature-constrained back-projection method for image super-resolution. Firstly, we take advantage of a feature-constrained polynomial interpolation method to enlarge the low-resolution image. Next, we consider low-frequency images of different resolutions to provide an example pair. Then, we use adaptive k NN search to find similar patches in the low-resolution image for every image patch in the high-resolution low-frequency image, leading to a regression model between similar patches to be learnt. The learnt model is applied to the low-resolution high-frequency image to produce high-resolution high-frequency information. An iterative back-projection algorithm is used as the final step to determine the final high-resolution image.Experimental results demonstrate that our method improves the visual quality of the high-resolution image.

  17. Human enamel structure studied by high resolution electron microscopy

    International Nuclear Information System (INIS)

    Wen, S.L.

    1989-01-01

    Human enamel structural features are characterized by high resolution electron microscopy. The human enamel consists of polycrystals with a structure similar to Ca10(PO4)6(OH)2. This article describes the structural features of human enamel crystal at atomic and nanometer level. Besides the structural description, a great number of high resolution images are included. Research into the carious process in human enamel is very important for human beings. This article firstly describes the initiation of caries in enamel crystal at atomic and unit-cell level and secondly describes the further steps of caries with structural and chemical demineralization. The demineralization in fact, is the origin of caries in human enamel. The remineralization of carious areas in human enamel has drawn more and more attention as its potential application is realized. This process has been revealed by high resolution electron microscopy in detail in this article. On the other hand, the radiation effects on the structure of human enamel are also characterized by high resolution electron microscopy. In order to reveal this phenomenon clearly, a great number of electron micrographs have been shown, and a physical mechanism is proposed. 26 references

  18. Restoration and Super-Resolution of Diffraction-Limited Imagery Data by Bayesian and Set-Theoretic Approaches

    National Research Council Canada - National Science Library

    Sundareshan, Malur

    2001-01-01

    This project was primarily aimed at the design of novel algorithms for the restoration and super-resolution processing of imagery data to improve the resolution in images acquired from practical sensing operations...

  19. Progress in high-resolution x-ray holographic microscopy

    International Nuclear Information System (INIS)

    Jacobsen, C.; Kirz, J.; Howells, M.; McQuaid, K.; Rothman, S.; Feder, R.; Sayre, D.

    1987-07-01

    Among the various types of x-ray microscopes that have been demonstrated, the holographic microscope has had the largest gap between promise and performance. The difficulties of fabricating x-ray optical elements have led some to view holography as the most attractive method for obtaining the ultimate in high resolution x-ray micrographs; however, we know of no investigations prior to 1987 that clearly demonstrated submicron resolution in reconstructed images. Previous efforts suffered from problems such as limited resolution and dynamic range in the recording media, low coherent x-ray flux, and aberrations and diffraction limits in visible light reconstruction. We have addressed the recording limitations through the use of an undulator x-ray source and high-resolution photoresist recording media. For improved results in the readout and reconstruction steps, we have employed metal shadowing and transmission electron microscopy, along with numerical reconstruction techniques. We believe that this approach will allow holography to emerge as a practical method of high-resolution x-ray microscopy. 30 refs., 4 figs

  20. Progress in high-resolution x-ray holographic microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Jacobsen, C.; Kirz, J.; Howells, M.; McQuaid, K.; Rothman, S.; Feder, R.; Sayre, D.

    1987-07-01

    Among the various types of x-ray microscopes that have been demonstrated, the holographic microscope has had the largest gap between promise and performance. The difficulties of fabricating x-ray optical elements have led some to view holography as the most attractive method for obtaining the ultimate in high resolution x-ray micrographs; however, we know of no investigations prior to 1987 that clearly demonstrated submicron resolution in reconstructed images. Previous efforts suffered from problems such as limited resolution and dynamic range in the recording media, low coherent x-ray flux, and aberrations and diffraction limits in visible light reconstruction. We have addressed the recording limitations through the use of an undulator x-ray source and high-resolution photoresist recording media. For improved results in the readout and reconstruction steps, we have employed metal shadowing and transmission electron microscopy, along with numerical reconstruction techniques. We believe that this approach will allow holography to emerge as a practical method of high-resolution x-ray microscopy. 30 refs., 4 figs.

  1. CO tip functionalization in subatomic resolution atomic force microscopy

    International Nuclear Information System (INIS)

    Kim, Minjung; Chelikowsky, James R.

    2015-01-01

    Noncontact atomic force microscopy (nc-AFM) employing a CO-functionalized tip displays dramatically enhanced resolution wherein covalent bonds of polycyclic aromatic hydrocarbon can be imaged. Employing real-space pseudopotential first-principles calculations, we examine the role of CO in functionalizing the nc-AFM tip. Our calculations allow us to simulate full AFM images and ascertain the enhancement mechanism of the CO molecule. We consider two approaches: one with an explicit inclusion of the CO molecule and one without. By comparing our simulations to existing experimental images, we ascribe the enhanced resolution of the CO functionalized tip to the special orbital characteristics of the CO molecule

  2. Optical super-resolution effect induced by nonlinear characteristics of graphene oxide films

    Science.gov (United States)

    Zhao, Yong-chuang; Nie, Zhong-quan; Zhai, Ai-ping; Tian, Yan-ting; Liu, Chao; Shi, Chang-kun; Jia, Bao-hua

    2018-01-01

    In this work, we focus on the optical super-resolution effect induced by strong nonlinear saturation absorption (NSA) of graphene oxide (GO) membranes. The third-order optical nonlinearities are characterized by the canonical Z-scan technique under femtosecond laser (wavelength: 800 nm, pulse width: 100 fs) excitation. Through controlling the applied femtosecond laser energy, NSA of the GO films can be tuned continuously. The GO film is placed at the focal plane as a unique amplitude filter to improve the resolution of the focused field. A multi-layer system model is proposed to present the generation of a deep sub-wavelength spot associated with the nonlinearity of GO films. Moreover, the parameter conditions to achieve the best resolution (˜λ/6) are determined entirely. The demonstrated results here are useful for high density optical recoding and storage, nanolithography, and super-resolution optical imaging.

  3. Single image super-resolution via regularized extreme learning regression for imagery from microgrid polarimeters

    Science.gov (United States)

    Sargent, Garrett C.; Ratliff, Bradley M.; Asari, Vijayan K.

    2017-08-01

    The advantage of division of focal plane imaging polarimeters is their ability to obtain temporally synchronized intensity measurements across a scene; however, they sacrifice spatial resolution in doing so due to their spatially modulated arrangement of the pixel-to-pixel polarizers and often result in aliased imagery. Here, we propose a super-resolution method based upon two previously trained extreme learning machines (ELM) that attempt to recover missing high frequency and low frequency content beyond the spatial resolution of the sensor. This method yields a computationally fast and simple way of recovering lost high and low frequency content from demosaicing raw microgrid polarimetric imagery. The proposed method outperforms other state-of-the-art single-image super-resolution algorithms in terms of structural similarity and peak signal-to-noise ratio.

  4. High-resolution and super stacking of time-reversal mirrors in locating seismic sources

    KAUST Repository

    Cao, Weiping

    2011-07-08

    Time reversal mirrors can be used to backpropagate and refocus incident wavefields to their actual source location, with the subsequent benefits of imaging with high-resolution and super-stacking properties. These benefits of time reversal mirrors have been previously verified with computer simulations and laboratory experiments but not with exploration-scale seismic data. We now demonstrate the high-resolution and the super-stacking properties in locating seismic sources with field seismic data that include multiple scattering. Tests on both synthetic data and field data show that a time reversal mirror has the potential to exceed the Rayleigh resolution limit by factors of 4 or more. Results also show that a time reversal mirror has a significant resilience to strong Gaussian noise and that accurate imaging of source locations from passive seismic data can be accomplished with traces having signal-to-noise ratios as low as 0.001. Synthetic tests also demonstrate that time reversal mirrors can sometimes enhance the signal by a factor proportional to the square root of the product of the number of traces, denoted as N and the number of events in the traces. This enhancement property is denoted as super-stacking and greatly exceeds the classical signal-to-noise enhancement factor of. High-resolution and super-stacking are properties also enjoyed by seismic interferometry and reverse-time migration with the exact velocity model. © 2011 European Association of Geoscientists & Engineers.

  5. High resolution FISH on super-stretched flow-sorted plant chromosomes.

    NARCIS (Netherlands)

    Valárik, M.; Bartos, J.; Kovarova, P.; Kubalakova, M.; Jong, de J.H.S.G.M.; Dolezel, J.

    2004-01-01

    A novel high-resolution fluorescence in situ hybridisation (FISH) strategy, using super-stretched flow-sorted plant chromosomes as targets, is described. The technique that allows longitudinal extension of chromosomes of more than 100 times their original metaphase size is especially attractive for

  6. Super-resolution links vinculin localization to function in focal adhesions.

    Science.gov (United States)

    Giannone, Grégory

    2015-07-01

    Integrin-based focal adhesions integrate biochemical and biomechanical signals from the extracellular matrix and the actin cytoskeleton. The combination of three-dimensional super-resolution imaging and loss- or gain-of-function protein mutants now links the nanoscale dynamic localization of proteins to their activation and function within focal adhesions.

  7. Low-Power Super-resolution Readout with Antimony Bismuth Alloy Film as Mask layer

    International Nuclear Information System (INIS)

    Lai-Xin, Jiang; Yi-Qun, Wu; Yang, Wang; Jing-Song, Wei; Fu-Xi, Gan

    2009-01-01

    Sb–Bi alloy films are proposed as a new kind of super-resolution mask layer with low readout threshold power. Using the Sb–Bi alloy film as a mask layer and SiN as a protective layer in a read-only memory disc, the super-resolution pits with diameters of 380 nm are read out by a dynamic setup, the laser wavelength is 780 nm and the numerical aperture of pickup lens is 0.45. The effects of the Sb–Bi thin film thickness, laser readout power and disc rotating velocity on the readout signal are investigated. The results show that the threshold laser power of super-resolution readout of the Sb–Bi mask layer is about 0.5 mW, and the corresponding carrier-to-noise ratio is about 20 dB at the film thickness of 50 nm. The super-resolution mechanism of the Sb–Bi alloy mask layer is discussed based on its temperature dependence of reflection

  8. A super-resolution approach for uncertainty estimation of PIV measurements

    NARCIS (Netherlands)

    Sciacchitano, A.; Wieneke, B.; Scarano, F.

    2012-01-01

    A super-resolution approach is proposed for the a posteriori uncertainty estimation of PIV measurements. The measured velocity field is employed to determine the displacement of individual particle images. A disparity set is built from the residual distance between paired particle images of

  9. High spatial resolution Kelvin probe force microscopy with coaxial probes

    International Nuclear Information System (INIS)

    Brown, Keith A; Westervelt, Robert M; Satzinger, Kevin J

    2012-01-01

    Kelvin probe force microscopy (KPFM) is a widely used technique to measure the local contact potential difference (CPD) between an AFM probe and the sample surface via the electrostatic force. The spatial resolution of KPFM is intrinsically limited by the long range of the electrostatic interaction, which includes contributions from the macroscopic cantilever and the conical tip. Here, we present coaxial AFM probes in which the cantilever and cone are shielded by a conducting shell, confining the tip–sample electrostatic interaction to a small region near the end of the tip. We have developed a technique to measure the true CPD despite the presence of the shell electrode. We find that the behavior of these probes agrees with an electrostatic model of the force, and we observe a factor of five improvement in spatial resolution relative to unshielded probes. Our discussion centers on KPFM, but the field confinement offered by these probes may improve any variant of electrostatic force microscopy. (paper)

  10. High spatial resolution soft-x-ray microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Meyer-Ilse, W.; Medecki, H.; Brown, J.T. [Ernest Orlando Lawrence Berkeley National Lab., CA (United States)] [and others

    1997-04-01

    A new soft x-ray microscope (XM-1) with high spatial resolution has been constructed by the Center for X-ray Optics. It uses bending magnet radiation from beamline 6.1 at the Advanced Light Source, and is used in a variety of projects and applications in the life and physical sciences. Most of these projects are ongoing. The instrument uses zone plate lenses and achieves a resolution of 43 nm, measured over 10% to 90% intensity with a knife edge test sample. X-ray microscopy permits the imaging of relatively thick samples, up to 10 {mu}m thick, in water. XM-1 has an easy to use interface, that utilizes visible light microscopy to precisely position and focus the specimen. The authors describe applications of this device in the biological sciences, as well as in studying industrial applications including structured polymer samples.

  11. Towards breaking the spatial resolution barriers: An optical flow and super-resolution approach for sea ice motion estimation

    Science.gov (United States)

    Petrou, Zisis I.; Xian, Yang; Tian, YingLi

    2018-04-01

    Estimation of sea ice motion at fine scales is important for a number of regional and local level applications, including modeling of sea ice distribution, ocean-atmosphere and climate dynamics, as well as safe navigation and sea operations. In this study, we propose an optical flow and super-resolution approach to accurately estimate motion from remote sensing images at a higher spatial resolution than the original data. First, an external example learning-based super-resolution method is applied on the original images to generate higher resolution versions. Then, an optical flow approach is applied on the higher resolution images, identifying sparse correspondences and interpolating them to extract a dense motion vector field with continuous values and subpixel accuracies. Our proposed approach is successfully evaluated on passive microwave, optical, and Synthetic Aperture Radar data, proving appropriate for multi-sensor applications and different spatial resolutions. The approach estimates motion with similar or higher accuracy than the original data, while increasing the spatial resolution of up to eight times. In addition, the adopted optical flow component outperforms a state-of-the-art pattern matching method. Overall, the proposed approach results in accurate motion vectors with unprecedented spatial resolutions of up to 1.5 km for passive microwave data covering the entire Arctic and 20 m for radar data, and proves promising for numerous scientific and operational applications.

  12. Multi-frame super-resolution with quality self-assessment for retinal fundus videos.

    Science.gov (United States)

    Köhler, Thomas; Brost, Alexander; Mogalle, Katja; Zhang, Qianyi; Köhler, Christiane; Michelson, Georg; Hornegger, Joachim; Tornow, Ralf P

    2014-01-01

    This paper proposes a novel super-resolution framework to reconstruct high-resolution fundus images from multiple low-resolution video frames in retinal fundus imaging. Natural eye movements during an examination are used as a cue for super-resolution in a robust maximum a-posteriori scheme. In order to compensate heterogeneous illumination on the fundus, we integrate retrospective illumination correction for photometric registration to the underlying imaging model. Our method utilizes quality self-assessment to provide objective quality scores for reconstructed images as well as to select regularization parameters automatically. In our evaluation on real data acquired from six human subjects with a low-cost video camera, the proposed method achieved considerable enhancements of low-resolution frames and improved noise and sharpness characteristics by 74%. In terms of image analysis, we demonstrate the importance of our method for the improvement of automatic blood vessel segmentation as an example application, where the sensitivity was increased by 13% using super-resolution reconstruction.

  13. Super-resolution processing for pulsed neutron imaging system using a high-speed camera

    International Nuclear Information System (INIS)

    Ishizuka, Ken; Kai, Tetsuya; Shinohara, Takenao; Segawa, Mariko; Mochiki, Koichi

    2015-01-01

    Super-resolution and center-of-gravity processing improve the resolution of neutron-transmitted images. These processing methods calculate the center-of-gravity pixel or sub-pixel of the neutron point converted into light by a scintillator. The conventional neutron-transmitted image is acquired using a high-speed camera by integrating many frames when a transmitted image with one frame is not provided. It succeeds in acquiring the transmitted image and calculating a spectrum by integrating frames of the same energy. However, because a high frame rate is required for neutron resonance absorption imaging, the number of pixels of the transmitted image decreases, and the resolution decreases to the limit of the camera performance. Therefore, we attempt to improve the resolution by integrating the frames after applying super-resolution or center-of-gravity processing. The processed results indicate that center-of-gravity processing can be effective in pulsed-neutron imaging with a high-speed camera. In addition, the results show that super-resolution processing is effective indirectly. A project to develop a real-time image data processing system has begun, and this system will be used at J-PARC in JAEA. (author)

  14. New learning based super-resolution: use of DWT and IGMRF prior.

    Science.gov (United States)

    Gajjar, Prakash P; Joshi, Manjunath V

    2010-05-01

    In this paper, we propose a new learning-based approach for super-resolving an image captured at low spatial resolution. Given the low spatial resolution test image and a database consisting of low and high spatial resolution images, we obtain super-resolution for the test image. We first obtain an initial high-resolution (HR) estimate by learning the high-frequency details from the available database. A new discrete wavelet transform (DWT) based approach is proposed for learning that uses a set of low-resolution (LR) images and their corresponding HR versions. Since the super-resolution is an ill-posed problem, we obtain the final solution using a regularization framework. The LR image is modeled as the aliased and noisy version of the corresponding HR image, and the aliasing matrix entries are estimated using the test image and the initial HR estimate. The prior model for the super-resolved image is chosen as an Inhomogeneous Gaussian Markov random field (IGMRF) and the model parameters are estimated using the same initial HR estimate. A maximum a posteriori (MAP) estimation is used to arrive at the cost function which is minimized using a simple gradient descent approach. We demonstrate the effectiveness of the proposed approach by conducting the experiments on gray scale as well as on color images. The method is compared with the standard interpolation technique and also with existing learning-based approaches. The proposed approach can be used in applications such as wildlife sensor networks, remote surveillance where the memory, the transmission bandwidth, and the camera cost are the main constraints.

  15. 3D super-resolution imaging with blinking quantum dots

    Science.gov (United States)

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

    2013-01-01

    Quantum dots are promising candidates for single molecule imaging due to their exceptional photophysical properties, including their intense brightness and resistance to photobleaching. They are also notorious for their blinking. Here we report a novel way to take advantage of quantum dot blinking to develop an imaging technique in three-dimensions with nanometric resolution. We first applied this method to simulated images of quantum dots, and then to quantum dots immobilized on microspheres. We achieved imaging resolutions (FWHM) of 8–17 nm in the x-y plane and 58 nm (on coverslip) or 81 nm (deep in solution) in the z-direction, approximately 3–7 times better than what has been achieved previously with quantum dots. This approach was applied to resolve the 3D distribution of epidermal growth factor receptor (EGFR) molecules at, and inside of, the plasma membrane of resting basal breast cancer cells. PMID:24093439

  16. Video-to-Video Dynamic Super-Resolution for Grayscale and Color Sequences

    Directory of Open Access Journals (Sweden)

    Elad Michael

    2006-01-01

    Full Text Available We address the dynamic super-resolution (SR problem of reconstructing a high-quality set of monochromatic or color super-resolved images from low-quality monochromatic, color, or mosaiced frames. Our approach includes a joint method for simultaneous SR, deblurring, and demosaicing, this way taking into account practical color measurements encountered in video sequences. For the case of translational motion and common space-invariant blur, the proposed method is based on a very fast and memory efficient approximation of the Kalman filter (KF. Experimental results on both simulated and real data are supplied, demonstrating the presented algorithms, and their strength.

  17. An Example-Based Super-Resolution Algorithm for Selfie Images

    Directory of Open Access Journals (Sweden)

    Jino Hans William

    2016-01-01

    Full Text Available A selfie is typically a self-portrait captured using the front camera of a smartphone. Most state-of-the-art smartphones are equipped with a high-resolution (HR rear camera and a low-resolution (LR front camera. As selfies are captured by front camera with limited pixel resolution, the fine details in it are explicitly missed. This paper aims to improve the resolution of selfies by exploiting the fine details in HR images captured by rear camera using an example-based super-resolution (SR algorithm. HR images captured by rear camera carry significant fine details and are used as an exemplar to train an optimal matrix-value regression (MVR operator. The MVR operator serves as an image-pair priori which learns the correspondence between the LR-HR patch-pairs and is effectively used to super-resolve LR selfie images. The proposed MVR algorithm avoids vectorization of image patch-pairs and preserves image-level information during both learning and recovering process. The proposed algorithm is evaluated for its efficiency and effectiveness both qualitatively and quantitatively with other state-of-the-art SR algorithms. The results validate that the proposed algorithm is efficient as it requires less than 3 seconds to super-resolve LR selfie and is effective as it preserves sharp details without introducing any counterfeit fine details.

  18. Super-resolution of facial images in forensics scenarios

    DEFF Research Database (Denmark)

    Satiro, Joao; Nasrollahi, Kamal; Correia, Paulo

    2015-01-01

    -resolution (SR) algorithms might be used. But, the problem with these algorithms is that they mostly require motion estimation between LR and low-quality images which is not always practical. To deal with this, we first simply interpolate the LR input images and then perform motion estimation. The estimated...... motion parameters are then used in a non-local mean-based SR algorithm to produce a higher quality image. This image is further fused with the interpolated version of the reference image via an alpha-blending approach. The experimental results on benchmark datasets and locally collected videos from...

  19. An angle encoder for super-high resolution and super-high accuracy using SelfA

    Science.gov (United States)

    Watanabe, Tsukasa; Kon, Masahito; Nabeshima, Nobuo; Taniguchi, Kayoko

    2014-06-01

    Angular measurement technology at high resolution for applications such as in hard disk drive manufacturing machines, precision measurement equipment and aspherical process machines requires a rotary encoder with high accuracy, high resolution and high response speed. However, a rotary encoder has angular deviation factors during operation due to scale error or installation error. It has been assumed to be impossible to achieve accuracy below 0.1″ in angular measurement or control after the installation onto the rotating axis. Self-calibration (Lu and Trumper 2007 CIRP Ann. 56 499; Kim et al 2011 Proc. MacroScale; Probst 2008 Meas. Sci. Technol. 19 015101; Probst et al Meas. Sci. Technol. 9 1059; Tadashi and Makoto 1993 J. Robot. Mechatronics 5 448; Ralf et al 2006 Meas. Sci. Technol. 17 2811) and cross-calibration (Probst et al 1998 Meas. Sci. Technol. 9 1059; Just et al 2009 Precis. Eng. 33 530; Burnashev 2013 Quantum Electron. 43 130) technologies for a rotary encoder have been actively discussed on the basis of the principle of circular closure. This discussion prompted the development of rotary tables which achieve reliable and high accuracy angular verification. We apply these technologies for the development of a rotary encoder not only to meet the requirement of super-high accuracy but also to meet that of super-high resolution. This paper presents the development of an encoder with 221 = 2097 152 resolutions per rotation (360°), that is, corresponding to a 0.62″ signal period, achieved by the combination of a laser rotary encoder supplied by Magnescale Co., Ltd and a self-calibratable encoder (SelfA) supplied by The National Institute of Advanced Industrial Science & Technology (AIST). In addition, this paper introduces the development of a rotary encoder to guarantee ±0.03″ accuracy at any point of the interpolated signal, with respect to the encoder at the minimum resolution of 233, that is, corresponding to a 0.0015″ signal period after

  20. An angle encoder for super-high resolution and super-high accuracy using SelfA

    International Nuclear Information System (INIS)

    Watanabe, Tsukasa; Kon, Masahito; Nabeshima, Nobuo; Taniguchi, Kayoko

    2014-01-01

    Angular measurement technology at high resolution for applications such as in hard disk drive manufacturing machines, precision measurement equipment and aspherical process machines requires a rotary encoder with high accuracy, high resolution and high response speed. However, a rotary encoder has angular deviation factors during operation due to scale error or installation error. It has been assumed to be impossible to achieve accuracy below 0.1″ in angular measurement or control after the installation onto the rotating axis. Self-calibration (Lu and Trumper 2007 CIRP Ann. 56 499; Kim et al 2011 Proc. MacroScale; Probst 2008 Meas. Sci. Technol. 19 015101; Probst et al Meas. Sci. Technol. 9 1059; Tadashi and Makoto 1993 J. Robot. Mechatronics 5 448; Ralf et al 2006 Meas. Sci. Technol. 17 2811) and cross-calibration (Probst et al 1998 Meas. Sci. Technol. 9 1059; Just et al 2009 Precis. Eng. 33 530; Burnashev 2013 Quantum Electron. 43 130) technologies for a rotary encoder have been actively discussed on the basis of the principle of circular closure. This discussion prompted the development of rotary tables which achieve reliable and high accuracy angular verification. We apply these technologies for the development of a rotary encoder not only to meet the requirement of super-high accuracy but also to meet that of super-high resolution. This paper presents the development of an encoder with 2 21 = 2097 152 resolutions per rotation (360°), that is, corresponding to a 0.62″ signal period, achieved by the combination of a laser rotary encoder supplied by Magnescale Co., Ltd and a self-calibratable encoder (SelfA) supplied by The National Institute of Advanced Industrial Science and Technology (AIST). In addition, this paper introduces the development of a rotary encoder to guarantee ±0.03″ accuracy at any point of the interpolated signal, with respect to the encoder at the minimum resolution of 2 33 , that is, corresponding to a 0.0015″ signal period

  1. Energy Resolution of the CMS ECAL Barrel Super-Module Using MGPA Electronics

    CERN Document Server

    Dewhirst, G

    2004-01-01

    A method for extracting the amplitude of a pulse has been developed in order to optimise the energy resolution of a CMS ECAL super-module in the 2003 test beam. The method consists of using a set of weights to extract the amplitude as well as subtract the baseline level event by event. For an incident beam impact restricted to an area of 4times 4~mm^2 the resolution of the ECAL, which uses the final prototype version of the MGPA electronics, is shown to match the target resolution.

  2. Super-Resolution Image Reconstruction Applied to Medical Ultrasound

    Science.gov (United States)

    Ellis, Michael

    Ultrasound is the preferred imaging modality for many diagnostic applications due to its real-time image reconstruction and low cost. Nonetheless, conventional ultrasound is not used in many applications because of limited spatial resolution and soft tissue contrast. Most commercial ultrasound systems reconstruct images using a simple delay-and-sum architecture on receive, which is fast and robust but does not utilize all information available in the raw data. Recently, more sophisticated image reconstruction methods have been developed that make use of far more information in the raw data to improve resolution and contrast. One such method is the Time-Domain Optimized Near-Field Estimator (TONE), which employs a maximum a priori estimation to solve a highly underdetermined problem, given a well-defined system model. TONE has been shown to significantly improve both the contrast and resolution of ultrasound images when compared to conventional methods. However, TONE's lack of robustness to variations from the system model and extremely high computational cost hinder it from being readily adopted in clinical scanners. This dissertation aims to reduce the impact of TONE's shortcomings, transforming it from an academic construct to a clinically viable image reconstruction algorithm. By altering the system model from a collection of individual hypothetical scatterers to a collection of weighted, diffuse regions, dTONE is able to achieve much greater robustness to modeling errors. A method for efficient parallelization of dTONE is presented that reduces reconstruction time by more than an order of magnitude with little loss in image fidelity. An alternative reconstruction algorithm, called qTONE, is also developed and is able to reduce reconstruction times by another two orders of magnitude while simultaneously improving image contrast. Each of these methods for improving TONE are presented, their limitations are explored, and all are used in concert to reconstruct in

  3. High-resolution electron microscopy and its applications.

    Science.gov (United States)

    Li, F H

    1987-12-01

    A review of research on high-resolution electron microscopy (HREM) carried out at the Institute of Physics, the Chinese Academy of Sciences, is presented. Apart from the direct observation of crystal and quasicrystal defects for some alloys, oxides, minerals, etc., and the structure determination for some minute crystals, an approximate image-contrast theory named pseudo-weak-phase object approximation (PWPOA), which shows the image contrast change with crystal thickness, is described. Within the framework of PWPOA, the image contrast of lithium ions in the crystal of R-Li2Ti3O7 has been observed. The usefulness of diffraction analysis techniques such as the direct method and Patterson method in HREM is discussed. Image deconvolution and resolution enhancement for weak-phase objects by use of the direct method are illustrated. In addition, preliminary results of image restoration for thick crystals are given.

  4. Nanometric depth resolution from multi-focal images in microscopy.

    Science.gov (United States)

    Dalgarno, Heather I C; Dalgarno, Paul A; Dada, Adetunmise C; Towers, Catherine E; Gibson, Gavin J; Parton, Richard M; Davis, Ilan; Warburton, Richard J; Greenaway, Alan H

    2011-07-06

    We describe a method for tracking the position of small features in three dimensions from images recorded on a standard microscope with an inexpensive attachment between the microscope and the camera. The depth-measurement accuracy of this method is tested experimentally on a wide-field, inverted microscope and is shown to give approximately 8 nm depth resolution, over a specimen depth of approximately 6 µm, when using a 12-bit charge-coupled device (CCD) camera and very bright but unresolved particles. To assess low-flux limitations a theoretical model is used to derive an analytical expression for the minimum variance bound. The approximations used in the analytical treatment are tested using numerical simulations. It is concluded that approximately 14 nm depth resolution is achievable with flux levels available when tracking fluorescent sources in three dimensions in live-cell biology and that the method is suitable for three-dimensional photo-activated localization microscopy resolution. Sub-nanometre resolution could be achieved with photon-counting techniques at high flux levels.

  5. High-resolution electron microscopy of advanced materials

    Energy Technology Data Exchange (ETDEWEB)

    Mitchell, T.E.; Kung, H.H.; Sickafus, K.E.; Gray, G.T. III; Field, R.D.; Smith, J.F. [Los Alamos National Lab., NM (United States). Materials Science and Technology Div.

    1997-11-01

    This final report chronicles a three-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). The High-Resolution Electron Microscopy Facility has doubled in size and tripled in quality since the beginning of the three-year period. The facility now includes a field-emission scanning electron microscope, a 100 kV field-emission scanning transmission electron microscope (FE-STEM), a 300 kV field-emission high-resolution transmission electron microscope (FE-HRTEM), and a 300 kV analytical transmission electron microscope. A new orientation imaging microscope is being installed. X-ray energy dispersive spectrometers for chemical analysis are available on all four microscopes; parallel electron energy loss spectrometers are operational on the FE-STEM and FE-HRTEM. These systems enable evaluation of local atomic bonding, as well as chemical composition in nanometer-scale regions. The FE-HRTEM has a point-to-point resolution of 1.6 {angstrom}, but the resolution can be pushed to its information limit of 1 {angstrom} by computer reconstruction of a focal series of images. HRTEM has been used to image the atomic structure of defects such as dislocations, grain boundaries, and interfaces in a variety of materials from superconductors and ferroelectrics to structural ceramics and intermetallics.

  6. In situ high-resolution thermal microscopy on integrated circuits.

    Science.gov (United States)

    Zhuo, Guan-Yu; Su, Hai-Ching; Wang, Hsien-Yi; Chan, Ming-Che

    2017-09-04

    The miniaturization of metal tracks in integrated circuits (ICs) can cause abnormal heat dissipation, resulting in electrostatic discharge, overvoltage breakdown, and other unwanted issues. Unfortunately, locating areas of abnormal heat dissipation is limited either by the spatial resolution or imaging acquisition speed of current thermal analytical techniques. A rapid, non-contact approach to the thermal imaging of ICs with sub-μm resolution could help to alleviate this issue. In this work, based on the intensity of the temperature-dependent two-photon fluorescence (TPF) of Rhodamine 6G (R6G) material, we developed a novel fast and non-invasive thermal microscopy with a sub-μm resolution. Its application to the location of hotspots that may evolve into thermally induced defects in ICs was also demonstrated. To the best of our knowledge, this is the first study to present high-resolution 2D thermal microscopic images of ICs, showing the generation, propagation, and distribution of heat during its operation. According to the demonstrated results, this scheme has considerable potential for future in situ hotspot analysis during the optimization stage of IC development.

  7. Super-resolution using a light inception layer in convolutional neural network

    Science.gov (United States)

    Mou, Qinyang; Guo, Jun

    2018-04-01

    Recently, several models based on CNN architecture have achieved great result on Single Image Super-Resolution (SISR) problem. In this paper, we propose an image super-resolution method (SR) using a light inception layer in convolutional network (LICN). Due to the strong representation ability of our well-designed inception layer that can learn richer representation with less parameters, we can build our model with shallow architecture that can reduce the effect of vanishing gradients problem and save computational costs. Our model strike a balance between computational speed and the quality of the result. Compared with state-of-the-art result, we produce comparable or better results with faster computational speed.

  8. Multiple-image hiding using super resolution reconstruction in high-frequency domains

    Science.gov (United States)

    Li, Xiao-Wei; Zhao, Wu-Xiang; Wang, Jun; Wang, Qiong-Hua

    2017-12-01

    In this paper, a robust multiple-image hiding method using the computer-generated integral imaging and the modified super-resolution reconstruction algorithm is proposed. In our work, the host image is first transformed into frequency domains by cellular automata (CA), to assure the quality of the stego-image, the secret images are embedded into the CA high-frequency domains. The proposed method has the following advantages: (1) robustness to geometric attacks because of the memory-distributed property of elemental images, (2) increasing quality of the reconstructed secret images as the scheme utilizes the modified super-resolution reconstruction algorithm. The simulation results show that the proposed multiple-image hiding method outperforms other similar hiding methods and is robust to some geometric attacks, e.g., Gaussian noise and JPEG compression attacks.

  9. Application of regularization technique in image super-resolution algorithm via sparse representation

    Science.gov (United States)

    Huang, De-tian; Huang, Wei-qin; Huang, Hui; Zheng, Li-xin

    2017-11-01

    To make use of the prior knowledge of the image more effectively and restore more details of the edges and structures, a novel sparse coding objective function is proposed by applying the principle of the non-local similarity and manifold learning on the basis of super-resolution algorithm via sparse representation. Firstly, the non-local similarity regularization term is constructed by using the similar image patches to preserve the edge information. Then, the manifold learning regularization term is constructed by utilizing the locally linear embedding approach to enhance the structural information. The experimental results validate that the proposed algorithm has a significant improvement compared with several super-resolution algorithms in terms of the subjective visual effect and objective evaluation indices.

  10. A simple model explaining super-resolution in absolute optical instruments

    Science.gov (United States)

    Leonhardt, Ulf; Sahebdivan, Sahar; Kogan, Alex; Tyc, Tomáš

    2015-05-01

    We develop a simple, one-dimensional model for super-resolution in absolute optical instruments that is able to describe the interplay between sources and detectors. Our model explains the subwavelength sensitivity of a point detector to a point source reported in previous computer simulations and experiments (Miñano 2011 New J. Phys.13 125009; Miñano 2014 New J. Phys.16 033015).

  11. Windowed time-reversal music technique for super-resolution ultrasound imaging

    Science.gov (United States)

    Huang, Lianjie; Labyed, Yassin

    2018-05-01

    Systems and methods for super-resolution ultrasound imaging using a windowed and generalized TR-MUSIC algorithm that divides the imaging region into overlapping sub-regions and applies the TR-MUSIC algorithm to the windowed backscattered ultrasound signals corresponding to each sub-region. The algorithm is also structured to account for the ultrasound attenuation in the medium and the finite-size effects of ultrasound transducer elements.

  12. MIiSR: Molecular Interactions in Super-Resolution Imaging Enables the Analysis of Protein Interactions, Dynamics and Formation of Multi-protein Structures.

    Directory of Open Access Journals (Sweden)

    Fabiana A Caetano

    2015-12-01

    Full Text Available Our current understanding of the molecular mechanisms which regulate cellular processes such as vesicular trafficking has been enabled by conventional biochemical and microscopy techniques. However, these methods often obscure the heterogeneity of the cellular environment, thus precluding a quantitative assessment of the molecular interactions regulating these processes. Herein, we present Molecular Interactions in Super Resolution (MIiSR software which provides quantitative analysis tools for use with super-resolution images. MIiSR combines multiple tools for analyzing intermolecular interactions, molecular clustering and image segmentation. These tools enable quantification, in the native environment of the cell, of molecular interactions and the formation of higher-order molecular complexes. The capabilities and limitations of these analytical tools are demonstrated using both modeled data and examples derived from the vesicular trafficking system, thereby providing an established and validated experimental workflow capable of quantitatively assessing molecular interactions and molecular complex formation within the heterogeneous environment of the cell.

  13. Super-resolution for a point source better than λ/500 using positive refraction

    International Nuclear Information System (INIS)

    Miñano, Juan C; González, Juan C; Benítez, Pablo; Grabovickic, Dejan; Marqués, Ricardo; Delgado, Vicente; Freire, Manuel

    2011-01-01

    Leonhardt (2009 New J. Phys. 11 093040) demonstrated that the two-dimensional (2D) Maxwell fish eye (MFE) lens can focus perfectly 2D Helmholtz waves of arbitrary frequency; that is, it can transport perfectly an outward (monopole) 2D Helmholtz wave field, generated by a point source, towards a ‘perfect point drain’ located at the corresponding image point. Moreover, a prototype with λ/5 super-resolution property for one microwave frequency has been manufactured and tested (Ma et al 2010 arXiv:1007.2530v1; Ma et al 2010 New J. Phys. 13 033016). However, neither software simulations nor experimental measurements for a broad band of frequencies have yet been reported. Here, we present steady-state simulations with a non-perfect drain for a device equivalent to the MFE, called the spherical geodesic waveguide (SGW), which predicts up to λ/500 super-resolution close to discrete frequencies. Out of these frequencies, the SGW does not show super-resolution in the analysis carried out. (paper)

  14. A Novel Method to Implement the Matrix Pencil Super Resolution Algorithm for Indoor Positioning

    Directory of Open Access Journals (Sweden)

    Tariq Jamil Saifullah Khanzada

    2011-10-01

    Full Text Available This article highlights the estimation of the results for the algorithms implemented in order to estimate the delays and distances for the indoor positioning system. The data sets for the transmitted and received signals are captured at a typical outdoor and indoor area. The estimation super resolution algorithms are applied. Different state of art and super resolution techniques based algorithms are applied to avail the optimal estimates of the delays and distances between the transmitted and received signals and a novel method for matrix pencil algorithm is devised. The algorithms perform variably at different scenarios of transmitted and received positions. Two scenarios are experienced, for the single antenna scenario the super resolution techniques like ESPRIT (Estimation of Signal Parameters via Rotational Invariance Technique and theMatrix Pencil algorithms give optimal performance compared to the conventional techniques. In two antenna scenario RootMUSIC and Matrix Pencil algorithm performed better than other algorithms for the distance estimation, however, the accuracy of all the algorithms is worst than the single antenna scenario. In all cases our devised Matrix Pencil algorithm achieved the best estimation results.

  15. Super-resolution for a point source better than λ/500 using positive refraction

    Science.gov (United States)

    Miñano, Juan C.; Marqués, Ricardo; González, Juan C.; Benítez, Pablo; Delgado, Vicente; Grabovickic, Dejan; Freire, Manuel

    2011-12-01

    Leonhardt (2009 New J. Phys. 11 093040) demonstrated that the two-dimensional (2D) Maxwell fish eye (MFE) lens can focus perfectly 2D Helmholtz waves of arbitrary frequency; that is, it can transport perfectly an outward (monopole) 2D Helmholtz wave field, generated by a point source, towards a ‘perfect point drain’ located at the corresponding image point. Moreover, a prototype with λ/5 super-resolution property for one microwave frequency has been manufactured and tested (Ma et al 2010 arXiv:1007.2530v1; Ma et al 2010 New J. Phys. 13 033016). However, neither software simulations nor experimental measurements for a broad band of frequencies have yet been reported. Here, we present steady-state simulations with a non-perfect drain for a device equivalent to the MFE, called the spherical geodesic waveguide (SGW), which predicts up to λ/500 super-resolution close to discrete frequencies. Out of these frequencies, the SGW does not show super-resolution in the analysis carried out.

  16. Large-area super-resolution optical imaging by using core-shell microfibers

    Science.gov (United States)

    Liu, Cheng-Yang; Lo, Wei-Chieh

    2017-09-01

    We first numerically and experimentally report large-area super-resolution optical imaging achieved by using core-shell microfibers. The particular spatial electromagnetic waves for different core-shell microfibers are studied by using finite-difference time-domain and ray tracing calculations. The focusing properties of photonic nanojets are evaluated in terms of intensity profile and full width at half-maximum along propagation and transversal directions. In experiment, the general optical fiber is chemically etched down to 6 μm diameter and coated with different metallic thin films by using glancing angle deposition. The direct imaging of photonic nanojets for different core-shell microfibers is performed with a scanning optical microscope system. We show that the intensity distribution of a photonic nanojet is highly related to the metallic shell due to the surface plasmon polaritons. Furthermore, large-area super-resolution optical imaging is performed by using different core-shell microfibers placed over the nano-scale grating with 150 nm line width. The core-shell microfiber-assisted imaging is achieved with super-resolution and hundreds of times the field-of-view in contrast to microspheres. The possible applications of these core-shell optical microfibers include real-time large-area micro-fluidics and nano-structure inspections.

  17. High resolution helium ion scanning microscopy of the rat kidney.

    Directory of Open Access Journals (Sweden)

    William L Rice

    Full Text Available Helium ion scanning microscopy is a novel imaging technology with the potential to provide sub-nanometer resolution images of uncoated biological tissues. So far, however, it has been used mainly in materials science applications. Here, we took advantage of helium ion microscopy to explore the epithelium of the rat kidney with unsurpassed image quality and detail. In addition, we evaluated different tissue preparation methods for their ability to preserve tissue architecture. We found that high contrast, high resolution imaging of the renal tubule surface is possible with a relatively simple processing procedure that consists of transcardial perfusion with aldehyde fixatives, vibratome tissue sectioning, tissue dehydration with graded methanol solutions and careful critical point drying. Coupled with the helium ion system, fine details such as membrane texture and membranous nanoprojections on the glomerular podocytes were visualized, and pores within the filtration slit diaphragm could be seen in much greater detail than in previous scanning EM studies. In the collecting duct, the extensive and striking apical microplicae of the intercalated cells were imaged without the shrunken or distorted appearance that is typical with conventional sample processing and scanning electron microscopy. Membrane depressions visible on principal cells suggest possible endo- or exocytotic events, and central cilia on these cells were imaged with remarkable preservation and clarity. We also demonstrate the use of colloidal gold probes for highlighting specific cell-surface proteins and find that 15 nm gold labels are practical and easily distinguishable, indicating that external labels of various sizes can be used to detect multiple targets in the same tissue. We conclude that this technology represents a technical breakthrough in imaging the topographical ultrastructure of animal tissues. Its use in future studies should allow the study of fine cellular details

  18. Development of the super high angular resolution principle for X-ray imaging

    International Nuclear Information System (INIS)

    Zhang Chen; Zhang Shuangnan

    2011-01-01

    Development of the Super High Angular Resolution Principle (SHARP) for coded-mask X-ray imaging is presented. We prove that SHARP can be considered as a generalized coded mask imaging method with a coding pattern comprised of diffraction-interference fringes in the mask pattern. The angular resolution of SHARP can be improved by detecting the fringes more precisely than the mask's element size, i.e. by using a detector with a pixel size smaller than the mask's element size. The proposed mission SHARP-X for solar X-ray observations is also briefly discussed. (research papers)

  19. Robust isotropic super-resolution by maximizing a Laplace posterior for MRI volumes

    Science.gov (United States)

    Han, Xian-Hua; Iwamoto, Yutaro; Shiino, Akihiko; Chen, Yen-Wei

    2014-03-01

    Magnetic resonance imaging can only acquire volume data with finite resolution due to various factors. In particular, the resolution in one direction (such as the slice direction) is much lower than others (such as the in-plane direction), yielding un-realistic visualizations. This study explores to reconstruct MRI isotropic resolution volumes from three orthogonal scans. This proposed super- resolution reconstruction is formulated as a maximum a posterior (MAP) problem, which relies on the generation model of the acquired scans from the unknown high-resolution volumes. Generally, the deviation ensemble of the reconstructed high-resolution (HR) volume from the available LR ones in the MAP is represented as a Gaussian distribution, which usually results in some noise and artifacts in the reconstructed HR volume. Therefore, this paper investigates a robust super-resolution by formulating the deviation set as a Laplace distribution, which assumes sparsity in the deviation ensemble based on the possible insight of the appeared large values only around some unexpected regions. In addition, in order to achieve reliable HR MRI volume, we integrates the priors such as bilateral total variation (BTV) and non-local mean (NLM) into the proposed MAP framework for suppressing artifacts and enriching visual detail. We validate the proposed robust SR strategy using MRI mouse data with high-definition resolution in two direction and low-resolution in one direction, which are imaged in three orthogonal scans: axial, coronal and sagittal planes. Experiments verifies that the proposed strategy can achieve much better HR MRI volumes than the conventional MAP method even with very high-magnification factor: 10.

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

    Science.gov (United States)

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

    2017-07-01

    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. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Super-resolution fluorescence imaging of nanoimprinted polymer patterns by selective fluorophore adsorption combined with redox switching

    KAUST Repository

    Yabiku, Y.; Kubo, S.; Nakagawa, M.; Vacha, M.; Habuchi, Satoshi

    2013-01-01

    We applied a super-resolution fluorescence imaging based on selective adsorption and redox switching of the fluorescent dye molecules for studying polymer nanostructures. We demonstrate that nano-scale structures of polymer thin films can

  2. Super-Resolution Reconstruction of Remote Sensing Images Using Multifractal Analysis

    Directory of Open Access Journals (Sweden)

    Mao-Gui Hu

    2009-10-01

    Full Text Available Satellite remote sensing (RS is an important contributor to Earth observation, providing various kinds of imagery every day, but low spatial resolution remains a critical bottleneck in a lot of applications, restricting higher spatial resolution analysis (e.g., intraurban. In this study, a multifractal-based super-resolution reconstruction method is proposed to alleviate this problem. The multifractal characteristic is common in Nature. The self-similarity or self-affinity presented in the image is useful to estimate details at larger and smaller scales than the original. We first look for the presence of multifractal characteristics in the images. Then we estimate parameters of the information transfer function and noise of the low resolution image. Finally, a noise-free, spatial resolutionenhanced image is generated by a fractal coding-based denoising and downscaling method. The empirical case shows that the reconstructed super-resolution image performs well indetail enhancement. This method is not only useful for remote sensing in investigating Earth, but also for other images with multifractal characteristics.

  3. Secondary electron spectroscopy and Auger microscopy at high spatial resolution. Application to scanning electron microscopy

    International Nuclear Information System (INIS)

    Le Gressus, Claude; Massignon, Daniel; Sopizet, Rene

    1979-01-01

    Secondary electron spectroscopy (SES), Auger electron spectroscopy (AES) and electron energy loss spectroscopy (ELS) are combined with ultra high vacuum scanning microscopy (SEM) for surface analysis at high spatial resolution. Reliability tests for the optical column for the vacuum and for the spectrometer are discussed. Furthermore the sensitivity threshold in AES which is compatible with a non destructive surface analysis at high spatial resolution is evaluated. This combination of all spectroscopies is used in the study of the beam damage correlated with the well known secondary electron image (SEI) darkening still observed in ultra high vacuum. The darkening is explained as a bulk decontamination of the sample rather than as a surface contamination from the residual vacuum gas [fr

  4. Development of a super-resolution optical microscope for directional dark matter search experiment

    International Nuclear Information System (INIS)

    Alexandrov, A.; Asada, T.; Consiglio, L.; D'Ambrosio, N.; De Lellis, G.; Di Crescenzo, A.; Di Marco, N.; Furuya, S.; Hakamata, K.; Ishikawa, M.; Katsuragawa, T.; Kuwabara, K.; Machii, S.; Naka, T.; Pupilli, F.; Sirignano, C.; Tawara, Y.; Tioukov, V.; Umemoto, A.; Yoshimoto, M.

    2016-01-01

    Nuclear emulsion is a perfect choice for a detector for directional DM search because of its high density and excellent position accuracy. The minimal detectable track length of a recoil nucleus in emulsion is required to be at least 100 nm, making the resolution of conventional optical microscopes insufficient to resolve them. Here we report about the R&D on a super-resolution optical microscope to be used in future directional DM search experiments with nuclear emulsion as a detector media. The microscope will be fully automatic, will use novel image acquisition and analysis techniques, will achieve the spatial resolution of the order of few tens of nm and will be capable of reconstructing recoil tracks with the length of at least 100 nm with high angular resolution.

  5. Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

    Science.gov (United States)

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

    2015-01-01

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

  6. Machine Learning Based Single-Frame Super-Resolution Processing for Lensless Blood Cell Counting

    Directory of Open Access Journals (Sweden)

    Xiwei Huang

    2016-11-01

    Full Text Available A lensless blood cell counting system integrating microfluidic channel and a complementary metal oxide semiconductor (CMOS image sensor is a promising technique to miniaturize the conventional optical lens based imaging system for point-of-care testing (POCT. However, such a system has limited resolution, making it imperative to improve resolution from the system-level using super-resolution (SR processing. Yet, how to improve resolution towards better cell detection and recognition with low cost of processing resources and without degrading system throughput is still a challenge. In this article, two machine learning based single-frame SR processing types are proposed and compared for lensless blood cell counting, namely the Extreme Learning Machine based SR (ELMSR and Convolutional Neural Network based SR (CNNSR. Moreover, lensless blood cell counting prototypes using commercial CMOS image sensors and custom designed backside-illuminated CMOS image sensors are demonstrated with ELMSR and CNNSR. When one captured low-resolution lensless cell image is input, an improved high-resolution cell image will be output. The experimental results show that the cell resolution is improved by 4×, and CNNSR has 9.5% improvement over the ELMSR on resolution enhancing performance. The cell counting results also match well with a commercial flow cytometer. Such ELMSR and CNNSR therefore have the potential for efficient resolution improvement in lensless blood cell counting systems towards POCT applications.

  7. Modeling noncontact atomic force microscopy resolution on corrugated surfaces

    Directory of Open Access Journals (Sweden)

    Kristen M. Burson

    2012-03-01

    Full Text Available Key developments in NC-AFM have generally involved atomically flat crystalline surfaces. However, many surfaces of technological interest are not atomically flat. We discuss the experimental difficulties in obtaining high-resolution images of rough surfaces, with amorphous SiO2 as a specific case. We develop a quasi-1-D minimal model for noncontact atomic force microscopy, based on van der Waals interactions between a spherical tip and the surface, explicitly accounting for the corrugated substrate (modeled as a sinusoid. The model results show an attenuation of the topographic contours by ~30% for tip distances within 5 Å of the surface. Results also indicate a deviation from the Hamaker force law for a sphere interacting with a flat surface.

  8. Advanced Microscopy of Microbial Cells

    DEFF Research Database (Denmark)

    Haagensen, Janus Anders Juul; Regenberg, Birgitte; Sternberg, Claus

    2011-01-01

    microscopy, super-resolution optical microscopy (STED, SIM, PALM) as well as atomic force microscopy and Raman spectroscopy. Using examples of bistability in microbial populations as well as biofilm development and differentiation in bacterial and yeast consortia, we demonstrate the importance of microscopy...

  9. Long-distance super-resolution imaging assisted by enhanced spatial Fourier transform.

    Science.gov (United States)

    Tang, Heng-He; Liu, Pu-Kun

    2015-09-07

    A new gradient-index (GRIN) lens that can realize enhanced spatial Fourier transform (FT) over optically long distances is demonstrated. By using an anisotropic GRIN metamaterial with hyperbolic dispersion, evanescent wave in free space can be transformed into propagating wave in the metamaterial and then focused outside due to negative-refraction. Both the results based on the ray tracing and the finite element simulation show that the spatial frequency bandwidth of the spatial FT can be extended to 2.7k(0) (k(0) is the wave vector in free space). Furthermore, assisted by the enhanced spatial FT, a new long-distance (in the optical far-field region) super-resolution imaging scheme is also proposed and the super resolved capability of λ/5 (λ is the wavelength in free space) is verified. The work may provide technical support for designing new-type high-speed microscopes with long working distances.

  10. Super Resolution and Interference Suppression Technique applied to SHARAD Radar Data

    Science.gov (United States)

    Raguso, M. C.; Mastrogiuseppe, M.; Seu, R.; Piazzo, L.

    2017-12-01

    We will present a super resolution and interference suppression technique applied to the data acquired by the SHAllow RADar (SHARAD) on board the NASA's 2005 Mars Reconnaissance Orbiter (MRO) mission, currently operating around Mars [1]. The algorithms allow to improve the range resolution roughly by a factor of 3 and the Signal to Noise Ratio (SNR) by a several decibels. Range compression algorithms usually adopt conventional Fourier transform techniques, which are limited in the resolution by the transmitted signal bandwidth, analogous to the Rayleigh's criterion in optics. In this work, we investigate a super resolution method based on autoregressive models and linear prediction techniques [2]. Starting from the estimation of the linear prediction coefficients from the spectral data, the algorithm performs the radar bandwidth extrapolation (BWE), thereby improving the range resolution of the pulse-compressed coherent radar data. Moreover, the EMIs (ElectroMagnetic Interferences) are detected and the spectra is interpolated in order to reconstruct an interference free spectrum, thereby improving the SNR. The algorithm can be applied to the single complex look image after synthetic aperture processing (SAR). We apply the proposed algorithm to simulated as well as to real radar data. We will demonstrate the effective enhancement on vertical resolution with respect to the classical spectral estimator. We will show that the imaging of the subsurface layered structures observed in radargrams is improved, allowing additional insights for the scientific community in the interpretation of the SHARAD radar data, which will help to further our understanding of the formation and evolution of known geological features on Mars. References: [1] Seu et al. 2007, Science, 2007, 317, 1715-1718 [2] K.M. Cuomo, "A Bandwidth Extrapolation Technique for Improved Range Resolution of Coherent Radar Data", Project Report CJP-60, Revision 1, MIT Lincoln Laboratory (4 Dec. 1992).

  11. High resolution, large deformation 3D traction force microscopy.

    Directory of Open Access Journals (Sweden)

    Jennet Toyjanova

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

  12. Observations of silicon carbide by high resolution transmission electron microscopy

    International Nuclear Information System (INIS)

    Smith, D.J.; Jepps, N.W.; Page, T.F.

    1978-01-01

    High resolution transmission electron microscopy techniques, principally involving direct lattice imaging, have been used as part of a study of the crystallography and phase transformation mechanics of silicon carbide polytypes. In particular, the 3C (cubic) and 6H (hexagonal) polytypes have been examined together with partially transformed structural mixtures. Although direct observation of two-dimensional atomic structures was not possible at an operating voltage of 100 kV, considerable microstructural information has been obtained by careful choice of the experimental conditions. In particular, tilted beam observations of the 0.25 nm lattice fringes have been made in the 3C polytype for two different brace 111 brace plane arrays in order to study the dimensions and coherency of finely-twinned regions together with brace 0006 brace and brace 1 0 bar1 2 brace lattice images in the 6H polytype which allow the detailed stacking operations to be resolved. Lower resolution lattice images formed with axial illumination have also been used to study the nature of the 3C → 6H transformation and results are presented showing that the transformation interface may originate with fine twinning of the 3C structure followed by growth of the resultant 6H regions. Observations have been made of the detailed stepped structure of this interface together with the stacking fault distribution in the resultant 6H material. (author)

  13. Super-resolution imaging of ESCRT-proteins at HIV-1 assembly sites.

    Directory of Open Access Journals (Sweden)

    Jens Prescher

    2015-02-01

    Full Text Available The cellular endosomal sorting complex required for transport (ESCRT machinery is involved in membrane budding processes, such as multivesicular biogenesis and cytokinesis. In HIV-infected cells, HIV-1 hijacks the ESCRT machinery to drive HIV release. Early in the HIV-1 assembly process, the ESCRT-I protein Tsg101 and the ESCRT-related protein ALIX are recruited to the assembly site. Further downstream, components such as the ESCRT-III proteins CHMP4 and CHMP2 form transient membrane associated lattices, which are involved in virus-host membrane fission. Although various geometries of ESCRT-III assemblies could be observed, the actual membrane constriction and fission mechanism is not fully understood. Fission might be driven from inside the HIV-1 budding neck by narrowing the membranes from the outside by larger lattices surrounding the neck, or from within the bud. Here, we use super-resolution fluorescence microscopy to elucidate the size and structure of the ESCRT components Tsg101, ALIX, CHMP4B and CHMP2A during HIV-1 budding below the diffraction limit. To avoid the deleterious effects of using fusion proteins attached to ESCRT components, we performed measurements on the endogenous protein or, in the case of CHMP4B, constructs modified with the small HA tag. Due to the transient nature of the ESCRT interactions, the fraction of HIV-1 assembly sites with colocalizing ESCRT complexes was low (1.5%-3.4%. All colocalizing ESCRT clusters exhibited closed, circular structures with an average size (full-width at half-maximum between 45 and 60 nm or a diameter (determined using a Ripley's L-function analysis of roughly 60 to 100 nm. The size distributions for colocalizing clusters were narrower than for non-colocalizing clusters, and significantly smaller than the HIV-1 bud. Hence, our results support a membrane scission process driven by ESCRT protein assemblies inside a confined structure, such as the bud neck, rather than by large lattices

  14. Effects of whispering gallery mode in microsphere super-resolution imaging

    Science.gov (United States)

    Zhou, Song; Deng, Yongbo; Zhou, Wenchao; Yu, Muxin; Urbach, H. P.; Wu, Yihui

    2017-09-01

    Whispering Gallery modes have been presented in microscopic glass spheres or toruses with many applications. In this paper, the possible approaches to enhance the imaging resolution by Whispering Gallery modes are discussed, including evanescent waves coupling, transformed and illustration by Whispering Gallery modes. It shows that the high-order scattering modes play the dominant role in the reconstructed virtual image when the Whispering Gallery modes exist. Furthermore, we find that the high image resolution of electric dipoles can be achieved, when the out-of-phase components exist from the illustration of Whispering Gallery modes. Those results of our simulation could contribute to the knowledge of microsphere-assisted super-resolution imaging and its potential applications.

  15. A Total Variation Regularization Based Super-Resolution Reconstruction Algorithm for Digital Video

    Directory of Open Access Journals (Sweden)

    Zhang Liangpei

    2007-01-01

    Full Text Available Super-resolution (SR reconstruction technique is capable of producing a high-resolution image from a sequence of low-resolution images. In this paper, we study an efficient SR algorithm for digital video. To effectively deal with the intractable problems in SR video reconstruction, such as inevitable motion estimation errors, noise, blurring, missing regions, and compression artifacts, the total variation (TV regularization is employed in the reconstruction model. We use the fixed-point iteration method and preconditioning techniques to efficiently solve the associated nonlinear Euler-Lagrange equations of the corresponding variational problem in SR. The proposed algorithm has been tested in several cases of motion and degradation. It is also compared with the Laplacian regularization-based SR algorithm and other TV-based SR algorithms. Experimental results are presented to illustrate the effectiveness of the proposed algorithm.

  16. Comparison between beamforming and super resolution imaging algorithms for non-destructive evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Chengguang [College of Mechatronic Engineering and Automation, National University of Defense Technology, Changsha 410073, PR China and Department of Mechanical Engineering, University of Bristol, Queen' s Building, University Walk, Bristol BS8 1TR (United Kingdom); Drinkwater, Bruce W. [Department of Mechanical Engineering, University of Bristol, Queen' s Building, University Walk, Bristol BS8 1TR (United Kingdom)

    2014-02-18

    In this paper the performance of total focusing method is compared with the widely used time-reversal MUSIC super resolution technique. The algorithms are tested with simulated and experimental ultrasonic array data, each containing different noise levels. The simulated time domain signals allow the effects of array geometry, frequency, scatterer location, scatterer size, scatterer separation and random noise to be carefully controlled. The performance of the imaging algorithms is evaluated in terms of resolution and sensitivity to random noise. It is shown that for the low noise situation, time-reversal MUSIC provides enhanced lateral resolution when compared to the total focusing method. However, for higher noise levels, the total focusing method shows robustness, whilst the performance of time-reversal MUSIC is significantly degraded.

  17. Comparison between beamforming and super resolution imaging algorithms for non-destructive evaluation

    International Nuclear Information System (INIS)

    Fan, Chengguang; Drinkwater, Bruce W.

    2014-01-01

    In this paper the performance of total focusing method is compared with the widely used time-reversal MUSIC super resolution technique. The algorithms are tested with simulated and experimental ultrasonic array data, each containing different noise levels. The simulated time domain signals allow the effects of array geometry, frequency, scatterer location, scatterer size, scatterer separation and random noise to be carefully controlled. The performance of the imaging algorithms is evaluated in terms of resolution and sensitivity to random noise. It is shown that for the low noise situation, time-reversal MUSIC provides enhanced lateral resolution when compared to the total focusing method. However, for higher noise levels, the total focusing method shows robustness, whilst the performance of time-reversal MUSIC is significantly degraded

  18. Droplet Image Super Resolution Based on Sparse Representation and Kernel Regression

    Science.gov (United States)

    Zou, Zhenzhen; Luo, Xinghong; Yu, Qiang

    2018-05-01

    Microgravity and containerless conditions, which are produced via electrostatic levitation combined with a drop tube, are important when studying the intrinsic properties of new metastable materials. Generally, temperature and image sensors can be used to measure the changes of sample temperature, morphology and volume. Then, the specific heat, surface tension, viscosity changes and sample density can be obtained. Considering that the falling speed of the material sample droplet is approximately 31.3 m/s when it reaches the bottom of a 50-meter-high drop tube, a high-speed camera with a collection rate of up to 106 frames/s is required to image the falling droplet. However, at the high-speed mode, very few pixels, approximately 48-120, will be obtained in each exposure time, which results in low image quality. Super-resolution image reconstruction is an algorithm that provides finer details than the sampling grid of a given imaging device by increasing the number of pixels per unit area in the image. In this work, we demonstrate the application of single image-resolution reconstruction in the microgravity and electrostatic levitation for the first time. Here, using the image super-resolution method based on sparse representation, a low-resolution droplet image can be reconstructed. Employed Yang's related dictionary model, high- and low-resolution image patches were combined with dictionary training, and high- and low-resolution-related dictionaries were obtained. The online double-sparse dictionary training algorithm was used in the study of related dictionaries and overcome the shortcomings of the traditional training algorithm with small image patch. During the stage of image reconstruction, the algorithm of kernel regression is added, which effectively overcomes the shortcomings of the Yang image's edge blurs.

  19. LFNet: A Novel Bidirectional Recurrent Convolutional Neural Network for Light-Field Image Super-Resolution.

    Science.gov (United States)

    Wang, Yunlong; Liu, Fei; Zhang, Kunbo; Hou, Guangqi; Sun, Zhenan; Tan, Tieniu

    2018-09-01

    The low spatial resolution of light-field image poses significant difficulties in exploiting its advantage. To mitigate the dependency of accurate depth or disparity information as priors for light-field image super-resolution, we propose an implicitly multi-scale fusion scheme to accumulate contextual information from multiple scales for super-resolution reconstruction. The implicitly multi-scale fusion scheme is then incorporated into bidirectional recurrent convolutional neural network, which aims to iteratively model spatial relations between horizontally or vertically adjacent sub-aperture images of light-field data. Within the network, the recurrent convolutions are modified to be more effective and flexible in modeling the spatial correlations between neighboring views. A horizontal sub-network and a vertical sub-network of the same network structure are ensembled for final outputs via stacked generalization. Experimental results on synthetic and real-world data sets demonstrate that the proposed method outperforms other state-of-the-art methods by a large margin in peak signal-to-noise ratio and gray-scale structural similarity indexes, which also achieves superior quality for human visual systems. Furthermore, the proposed method can enhance the performance of light field applications such as depth estimation.

  20. Super-Resolution Imaging of Protein Secretion Systems and the Cell Surface of Gram-Negative Bacteria

    Directory of Open Access Journals (Sweden)

    Sachith D. Gunasinghe

    2017-05-01

    Full Text Available Gram-negative bacteria have a highly evolved cell wall with two membranes composed of complex arrays of integral and peripheral proteins, as well as phospholipids and glycolipids. In order to sense changes in, respond to, and exploit their environmental niches, bacteria rely on structures assembled into or onto the outer membrane. Protein secretion across the cell wall is a key process in virulence and other fundamental aspects of bacterial cell biology. The final stage of protein secretion in Gram-negative bacteria, translocation across the outer membrane, is energetically challenging so sophisticated nanomachines have evolved to meet this challenge. Advances in fluorescence microscopy now allow for the direct visualization of the protein secretion process, detailing the dynamics of (i outer membrane biogenesis and the assembly of protein secretion systems into the outer membrane, (ii the spatial distribution of these and other membrane proteins on the bacterial cell surface, and (iii translocation of effector proteins, toxins and enzymes by these protein secretion systems. Here we review the frontier research imaging the process of secretion, particularly new studies that are applying various modes of super-resolution microscopy.

  1. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy.

    Science.gov (United States)

    Schulz, Olaf; Pieper, Christoph; Clever, Michaela; Pfaff, Janine; Ruhlandt, Aike; Kehlenbach, Ralph H; Wouters, Fred S; Großhans, Jörg; Bunt, Gertrude; Enderlein, Jörg

    2013-12-24

    We demonstrate how a conventional confocal spinning-disk (CSD) microscope can be converted into a doubly resolving image scanning microscopy (ISM) system without changing any part of its optical or mechanical elements. Making use of the intrinsic properties of a CSD microscope, we illuminate stroboscopically, generating an array of excitation foci that are moved across the sample by varying the phase between stroboscopic excitation and rotation of the spinning disk. ISM then generates an image with nearly doubled resolution. Using conventional fluorophores, we have imaged single nuclear pore complexes in the nuclear membrane and aggregates of GFP-conjugated Tau protein in three dimensions. Multicolor ISM was shown on cytoskeletal-associated structural proteins and on 3D four-color images including MitoTracker and Hoechst staining. The simple adaptation of conventional CSD equipment allows superresolution investigations of a broad variety of cell biological questions.

  2. Ordinal Regression Based Subpixel Shift Estimation for Video Super-Resolution

    Directory of Open Access Journals (Sweden)

    Petrovic Nemanja

    2007-01-01

    Full Text Available We present a supervised learning-based approach for subpixel motion estimation which is then used to perform video super-resolution. The novelty of this work is the formulation of the problem of subpixel motion estimation in a ranking framework. The ranking formulation is a variant of classification and regression formulation, in which the ordering present in class labels namely, the shift between patches is explicitly taken into account. Finally, we demonstrate the applicability of our approach on superresolving synthetically generated images with global subpixel shifts and enhancing real video frames by accounting for both local integer and subpixel shifts.

  3. Accelerating cross-validation with total variation and its application to super-resolution imaging.

    Directory of Open Access Journals (Sweden)

    Tomoyuki Obuchi

    Full Text Available We develop an approximation formula for the cross-validation error (CVE of a sparse linear regression penalized by ℓ1-norm and total variation terms, which is based on a perturbative expansion utilizing the largeness of both the data dimensionality and the model. The developed formula allows us to reduce the necessary computational cost of the CVE evaluation significantly. The practicality of the formula is tested through application to simulated black-hole image reconstruction on the event-horizon scale with super resolution. The results demonstrate that our approximation reproduces the CVE values obtained via literally conducted cross-validation with reasonably good precision.

  4. Time reversal and phase coherent music techniques for super-resolution ultrasound imaging

    Science.gov (United States)

    Huang, Lianjie; Labyed, Yassin

    2018-05-01

    Systems and methods for super-resolution ultrasound imaging using a windowed and generalized TR-MUSIC algorithm that divides the imaging region into overlapping sub-regions and applies the TR-MUSIC algorithm to the windowed backscattered ultrasound signals corresponding to each sub-region. The algorithm is also structured to account for the ultrasound attenuation in the medium and the finite-size effects of ultrasound transducer elements. A modified TR-MUSIC imaging algorithm is used to account for ultrasound scattering from both density and compressibility contrasts. The phase response of ultrasound transducer elements is accounted for in a PC-MUSIC system.

  5. Super-Resolution of Plant Disease Images for the Acceleration of Image-based Phenotyping and Vigor Diagnosis in Agriculture.

    Science.gov (United States)

    Yamamoto, Kyosuke; Togami, Takashi; Yamaguchi, Norio

    2017-11-06

    Unmanned aerial vehicles (UAVs or drones) are a very promising branch of technology, and they have been utilized in agriculture-in cooperation with image processing technologies-for phenotyping and vigor diagnosis. One of the problems in the utilization of UAVs for agricultural purposes is the limitation in flight time. It is necessary to fly at a high altitude to capture the maximum number of plants in the limited time available, but this reduces the spatial resolution of the captured images. In this study, we applied a super-resolution method to the low-resolution images of tomato diseases to recover detailed appearances, such as lesions on plant organs. We also conducted disease classification using high-resolution, low-resolution, and super-resolution images to evaluate the effectiveness of super-resolution methods in disease classification. Our results indicated that the super-resolution method outperformed conventional image scaling methods in spatial resolution enhancement of tomato disease images. The results of disease classification showed that the accuracy attained was also better by a large margin with super-resolution images than with low-resolution images. These results indicated that our approach not only recovered the information lost in low-resolution images, but also exerted a beneficial influence on further image analysis. The proposed approach will accelerate image-based phenotyping and vigor diagnosis in the field, because it not only saves time to capture images of a crop in a cultivation field but also secures the accuracy of these images for further analysis.

  6. Super-Resolution of Plant Disease Images for the Acceleration of Image-based Phenotyping and Vigor Diagnosis in Agriculture

    Directory of Open Access Journals (Sweden)

    Kyosuke Yamamoto

    2017-11-01

    Full Text Available Unmanned aerial vehicles (UAVs or drones are a very promising branch of technology, and they have been utilized in agriculture—in cooperation with image processing technologies—for phenotyping and vigor diagnosis. One of the problems in the utilization of UAVs for agricultural purposes is the limitation in flight time. It is necessary to fly at a high altitude to capture the maximum number of plants in the limited time available, but this reduces the spatial resolution of the captured images. In this study, we applied a super-resolution method to the low-resolution images of tomato diseases to recover detailed appearances, such as lesions on plant organs. We also conducted disease classification using high-resolution, low-resolution, and super-resolution images to evaluate the effectiveness of super-resolution methods in disease classification. Our results indicated that the super-resolution method outperformed conventional image scaling methods in spatial resolution enhancement of tomato disease images. The results of disease classification showed that the accuracy attained was also better by a large margin with super-resolution images than with low-resolution images. These results indicated that our approach not only recovered the information lost in low-resolution images, but also exerted a beneficial influence on further image analysis. The proposed approach will accelerate image-based phenotyping and vigor diagnosis in the field, because it not only saves time to capture images of a crop in a cultivation field but also secures the accuracy of these images for further analysis.

  7. Super resolution PLIF demonstrated in turbulent jet flows seeded with I2

    Science.gov (United States)

    Xu, Wenjiang; Liu, Ning; Ma, Lin

    2018-05-01

    Planar laser induced fluorescence (PLIF) represents an indispensable tool for flow and flame imaging. However, the PLIF technique suffers from limited spatial resolution or blurring in many situations, which restricts its applicability and capability. This work describes a new method, named SR-PLIF (super-resolution PLIF), to overcome these limitations and enhance the capability of PLIF. The method uses PLIF images captured simultaneously from two (or more) orientations to reconstruct a final PLIF image with resolution enhanced or blurring removed. This paper reports the development of the reconstruction algorithm, and the experimental demonstration of the SR-PLIF method both with controlled samples and with turbulent flows seeded with iodine vapor. Using controlled samples with two cameras, the spatial resolution in the best case was improved from 0.06 mm in the projections to 0.03 mm in the SR image, in terms of the spreading width of a sharp edge. With turbulent flows, an image sharpness measure was developed to quantify the spatial resolution, and SR reconstruction with two cameras can effectively improve the spatial resolution compared to the projections in terms of the sharpness measure.

  8. Detecting breast microcalcifications using super-resolution and wave-equation ultrasound imaging: a numerical phantom study

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Lianjie [Los Alamos National Laboratory; Simonetti, Francesco [IMPERIAL COLLEGE LONDON; Huthwaite, Peter [IMPERIAL COLLEGE LONDON; Rosenberg, Robert [UNM; Williamson, Michael [UNM

    2010-01-01

    Ultrasound image resolution and quality need to be significantly improved for breast microcalcification detection. Super-resolution imaging with the factorization method has recently been developed as a promising tool to break through the resolution limit of conventional imaging. In addition, wave-equation reflection imaging has become an effective method to reduce image speckles by properly handling ultrasound scattering/diffraction from breast heterogeneities during image reconstruction. We explore the capabilities of a novel super-resolution ultrasound imaging method and a wave-equation reflection imaging scheme for detecting breast microcalcifications. Super-resolution imaging uses the singular value decomposition and a factorization scheme to achieve an image resolution that is not possible for conventional ultrasound imaging. Wave-equation reflection imaging employs a solution to the acoustic-wave equation in heterogeneous media to backpropagate ultrasound scattering/diffraction waves to scatters and form images of heterogeneities. We construct numerical breast phantoms using in vivo breast images, and use a finite-difference wave-equation scheme to generate ultrasound data scattered from inclusions that mimic microcalcifications. We demonstrate that microcalcifications can be detected at full spatial resolution using the super-resolution ultrasound imaging and wave-equation reflection imaging methods.

  9. A theoretical investigation of super-resolution CARS imaging via coherent and incoherent saturation of transitions

    NARCIS (Netherlands)

    Beeker, W.P.; Beeker, Willem; Lee, Christopher James; Boller, Klaus J.; Gross, Petra; Gross, P.; Cleff, Carsten; Fallnich, Carsten; Offerhaus, Herman L.; Herek, Jennifer Lynn

    2011-01-01

    We review two approaches to achieving sub-diffraction-limited resolution coherent anti-Stokes Raman scattering (CARS) microscopy (Beeker et al., Opt. Express, 2009, 17, 22632 and Beeker et al., J. Herek, Phys. Rev. A, 2010, 81, 012507). We performed a numerical investigation, based on the density

  10. Single Image Super-Resolution Using Global Regression Based on Multiple Local Linear Mappings.

    Science.gov (United States)

    Choi, Jae-Seok; Kim, Munchurl

    2017-03-01

    Super-resolution (SR) has become more vital, because of its capability to generate high-quality ultra-high definition (UHD) high-resolution (HR) images from low-resolution (LR) input images. Conventional SR methods entail high computational complexity, which makes them difficult to be implemented for up-scaling of full-high-definition input images into UHD-resolution images. Nevertheless, our previous super-interpolation (SI) method showed a good compromise between Peak-Signal-to-Noise Ratio (PSNR) performances and computational complexity. However, since SI only utilizes simple linear mappings, it may fail to precisely reconstruct HR patches with complex texture. In this paper, we present a novel SR method, which inherits the large-to-small patch conversion scheme from SI but uses global regression based on local linear mappings (GLM). Thus, our new SR method is called GLM-SI. In GLM-SI, each LR input patch is divided into 25 overlapped subpatches. Next, based on the local properties of these subpatches, 25 different local linear mappings are applied to the current LR input patch to generate 25 HR patch candidates, which are then regressed into one final HR patch using a global regressor. The local linear mappings are learned cluster-wise in our off-line training phase. The main contribution of this paper is as follows: Previously, linear-mapping-based conventional SR methods, including SI only used one simple yet coarse linear mapping to each patch to reconstruct its HR version. On the contrary, for each LR input patch, our GLM-SI is the first to apply a combination of multiple local linear mappings, where each local linear mapping is found according to local properties of the current LR patch. Therefore, it can better approximate nonlinear LR-to-HR mappings for HR patches with complex texture. Experiment results show that the proposed GLM-SI method outperforms most of the state-of-the-art methods, and shows comparable PSNR performance with much lower

  11. Super-resolution pupil filtering for visual performance enhancement using adaptive optics

    Science.gov (United States)

    Zhao, Lina; Dai, Yun; Zhao, Junlei; Zhou, Xiaojun

    2018-05-01

    Ocular aberration correction can significantly improve visual function of the human eye. However, even under ideal aberration correction conditions, pupil diffraction restricts the resolution of retinal images. Pupil filtering is a simple super-resolution (SR) method that can overcome this diffraction barrier. In this study, a 145-element piezoelectric deformable mirror was used as a pupil phase filter because of its programmability and high fitting accuracy. Continuous phase-only filters were designed based on Zernike polynomial series and fitted through closed-loop adaptive optics. SR results were validated using double-pass point spread function images. Contrast sensitivity was further assessed to verify the SR effect on visual function. An F-test was conducted for nested models to statistically compare different CSFs. These results indicated CSFs for the proposed SR filter were significantly higher than the diffraction correction (p vision optical correction of the human eye.

  12. Remote classification from an airborne camera using image super-resolution.

    Science.gov (United States)

    Woods, Matthew; Katsaggelos, Aggelos

    2017-02-01

    The image processing technique known as super-resolution (SR), which attempts to increase the effective pixel sampling density of a digital imager, has gained rapid popularity over the last decade. The majority of literature focuses on its ability to provide results that are visually pleasing to a human observer. In this paper, we instead examine the ability of SR to improve the resolution-critical capability of an imaging system to perform a classification task from a remote location, specifically from an airborne camera. In order to focus the scope of the study, we address and quantify results for the narrow case of text classification. However, we expect the results generalize to a large set of related, remote classification tasks. We generate theoretical results through simulation, which are corroborated by experiments with a camera mounted on a DJI Phantom 3 quadcopter.

  13. Single image super-resolution based on approximated Heaviside functions and iterative refinement

    Science.gov (United States)

    Wang, Xin-Yu; Huang, Ting-Zhu; Deng, Liang-Jian

    2018-01-01

    One method of solving the single-image super-resolution problem is to use Heaviside functions. This has been done previously by making a binary classification of image components as “smooth” and “non-smooth”, describing these with approximated Heaviside functions (AHFs), and iteration including l1 regularization. We now introduce a new method in which the binary classification of image components is extended to different degrees of smoothness and non-smoothness, these components being represented by various classes of AHFs. Taking into account the sparsity of the non-smooth components, their coefficients are l1 regularized. In addition, to pick up more image details, the new method uses an iterative refinement for the residuals between the original low-resolution input and the downsampled resulting image. Experimental results showed that the new method is superior to the original AHF method and to four other published methods. PMID:29329298

  14. MAP-MRF-Based Super-Resolution Reconstruction Approach for Coded Aperture Compressive Temporal Imaging

    Directory of Open Access Journals (Sweden)

    Tinghua Zhang

    2018-02-01

    Full Text Available Coded Aperture Compressive Temporal Imaging (CACTI can afford low-cost temporal super-resolution (SR, but limits are imposed by noise and compression ratio on reconstruction quality. To utilize inter-frame redundant information from multiple observations and sparsity in multi-transform domains, a robust reconstruction approach based on maximum a posteriori probability and Markov random field (MAP-MRF model for CACTI is proposed. The proposed approach adopts a weighted 3D neighbor system (WNS and the coordinate descent method to perform joint estimation of model parameters, to achieve the robust super-resolution reconstruction. The proposed multi-reconstruction algorithm considers both total variation (TV and ℓ 2 , 1 norm in wavelet domain to address the minimization problem for compressive sensing, and solves it using an accelerated generalized alternating projection algorithm. The weighting coefficient for different regularizations and frames is resolved by the motion characteristics of pixels. The proposed approach can provide high visual quality in the foreground and background of a scene simultaneously and enhance the fidelity of the reconstruction results. Simulation results have verified the efficacy of our new optimization framework and the proposed reconstruction approach.

  15. Hyperspectral Super-Resolution of Locally Low Rank Images From Complementary Multisource Data.

    Science.gov (United States)

    Veganzones, Miguel A; Simoes, Miguel; Licciardi, Giorgio; Yokoya, Naoto; Bioucas-Dias, Jose M; Chanussot, Jocelyn

    2016-01-01

    Remote sensing hyperspectral images (HSIs) are quite often low rank, in the sense that the data belong to a low dimensional subspace/manifold. This has been recently exploited for the fusion of low spatial resolution HSI with high spatial resolution multispectral images in order to obtain super-resolution HSI. Most approaches adopt an unmixing or a matrix factorization perspective. The derived methods have led to state-of-the-art results when the spectral information lies in a low-dimensional subspace/manifold. However, if the subspace/manifold dimensionality spanned by the complete data set is large, i.e., larger than the number of multispectral bands, the performance of these methods mainly decreases because the underlying sparse regression problem is severely ill-posed. In this paper, we propose a local approach to cope with this difficulty. Fundamentally, we exploit the fact that real world HSIs are locally low rank, that is, pixels acquired from a given spatial neighborhood span a very low-dimensional subspace/manifold, i.e., lower or equal than the number of multispectral bands. Thus, we propose to partition the image into patches and solve the data fusion problem independently for each patch. This way, in each patch the subspace/manifold dimensionality is low enough, such that the problem is not ill-posed anymore. We propose two alternative approaches to define the hyperspectral super-resolution through local dictionary learning using endmember induction algorithms. We also explore two alternatives to define the local regions, using sliding windows and binary partition trees. The effectiveness of the proposed approaches is illustrated with synthetic and semi real data.

  16. Image Super-Resolution Algorithm Based on an Improved Sparse Autoencoder

    Directory of Open Access Journals (Sweden)

    Detian Huang

    2018-01-01

    Full Text Available Due to the limitations of the resolution of the imaging system and the influence of scene changes and other factors, sometimes only low-resolution images can be acquired, which cannot satisfy the practical application’s requirements. To improve the quality of low-resolution images, a novel super-resolution algorithm based on an improved sparse autoencoder is proposed. Firstly, in the training set preprocessing stage, the high- and low-resolution image training sets are constructed, respectively, by using high-frequency information of the training samples as the characterization, and then the zero-phase component analysis whitening technique is utilized to decorrelate the formed joint training set to reduce its redundancy. Secondly, a constructed sparse regularization term is added to the cost function of the traditional sparse autoencoder to further strengthen the sparseness constraint on the hidden layer. Finally, in the dictionary learning stage, the improved sparse autoencoder is adopted to achieve unsupervised dictionary learning to improve the accuracy and stability of the dictionary. Experimental results validate that the proposed algorithm outperforms the existing algorithms both in terms of the subjective visual perception and the objective evaluation indices, including the peak signal-to-noise ratio and the structural similarity measure.

  17. Spatiotemporal Super-Resolution Reconstruction Based on Robust Optical Flow and Zernike Moment for Video Sequences

    Directory of Open Access Journals (Sweden)

    Meiyu Liang

    2013-01-01

    Full Text Available In order to improve the spatiotemporal resolution of the video sequences, a novel spatiotemporal super-resolution reconstruction model (STSR based on robust optical flow and Zernike moment is proposed in this paper, which integrates the spatial resolution reconstruction and temporal resolution reconstruction into a unified framework. The model does not rely on accurate estimation of subpixel motion and is robust to noise and rotation. Moreover, it can effectively overcome the problems of hole and block artifacts. First we propose an efficient robust optical flow motion estimation model based on motion details preserving, then we introduce the biweighted fusion strategy to implement the spatiotemporal motion compensation. Next, combining the self-adaptive region correlation judgment strategy, we construct a fast fuzzy registration scheme based on Zernike moment for better STSR with higher efficiency, and then the final video sequences with high spatiotemporal resolution can be obtained by fusion of the complementary and redundant information with nonlocal self-similarity between the adjacent video frames. Experimental results demonstrate that the proposed method outperforms the existing methods in terms of both subjective visual and objective quantitative evaluations.

  18. High resolution magnetic force microscopy using focussed ion beam modified tips

    NARCIS (Netherlands)

    Phillips, G.N.; Siekman, Martin Herman; Abelmann, Leon; Lodder, J.C.

    2002-01-01

    Summary form only given. Magnetic force microscopy (MFM) is well established for imaging surface magnetic stray fields. With commercial microscopes and magnetic tips, images with 50 nm resolution are quite routine; however, obtaining higher resolutions is experimentally more demanding. Higher

  19. Super-resolution imaging of a 2.5 kb non-repetitive DNA in situ in the nuclear genome using molecular beacon probes

    Science.gov (United States)

    Ni, Yanxiang; Cao, Bo; Ma, Tszshan; Niu, Gang; Huo, Yingdong; Huang, Jiandong; Chen, Danni; Liu, Yi; Yu, Bin; Zhang, Michael Q; Niu, Hanben

    2017-01-01

    High-resolution visualization of short non-repetitive DNA in situ in the nuclear genome is essential for studying looping interactions and chromatin organization in single cells. Recent advances in fluorescence in situ hybridization (FISH) using Oligopaint probes have enabled super-resolution imaging of genomic domains with a resolution limit of 4.9 kb. To target shorter elements, we developed a simple FISH method that uses molecular beacon (MB) probes to facilitate the probe-target binding, while minimizing non-specific fluorescence. We used three-dimensional stochastic optical reconstruction microscopy (3D-STORM) with optimized imaging conditions to efficiently distinguish sparsely distributed Alexa-647 from background cellular autofluorescence. Utilizing 3D-STORM and only 29–34 individual MB probes, we observed 3D fine-scale nanostructures of 2.5 kb integrated or endogenous unique DNA in situ in human or mouse genome, respectively. We demonstrated our MB-based FISH method was capable of visualizing the so far shortest non-repetitive genomic sequence in 3D at super-resolution. DOI: http://dx.doi.org/10.7554/eLife.21660.001 PMID:28485713

  20. A novel algorithm of super-resolution image reconstruction based on multi-class dictionaries for natural scene

    Science.gov (United States)

    Wu, Wei; Zhao, Dewei; Zhang, Huan

    2015-12-01

    Super-resolution image reconstruction is an effective method to improve the image quality. It has important research significance in the field of image processing. However, the choice of the dictionary directly affects the efficiency of image reconstruction. A sparse representation theory is introduced into the problem of the nearest neighbor selection. Based on the sparse representation of super-resolution image reconstruction method, a super-resolution image reconstruction algorithm based on multi-class dictionary is analyzed. This method avoids the redundancy problem of only training a hyper complete dictionary, and makes the sub-dictionary more representatives, and then replaces the traditional Euclidean distance computing method to improve the quality of the whole image reconstruction. In addition, the ill-posed problem is introduced into non-local self-similarity regularization. Experimental results show that the algorithm is much better results than state-of-the-art algorithm in terms of both PSNR and visual perception.

  1. Localization microscopy of DNA in situ using Vybrant{sup ®} DyeCycle™ Violet fluorescent probe: A new approach to study nuclear nanostructure at single molecule resolution

    Energy Technology Data Exchange (ETDEWEB)

    Żurek-Biesiada, Dominika [Laboratory of Cell Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków (Poland); Szczurek, Aleksander T. [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Prakash, Kirti [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Institute for Pharmacy and Molecular Biotechnology (IPMB), University of Heidelberg, Im Neuenheimer Feld 364, D-69120 Heidelberg (Germany); Mohana, Giriram K. [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Lee, Hyun-Keun [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Department of Physics, University of Mainz (JGU), Staudingerweg 7, 55128 Mainz (Germany); Roignant, Jean-Yves [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Birk, Udo J. [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Department of Physics, University of Mainz (JGU), Staudingerweg 7, 55128 Mainz (Germany); Dobrucki, Jurek W., E-mail: jerzy.dobrucki@uj.edu.pl [Laboratory of Cell Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Kraków (Poland); Cremer, Christoph, E-mail: c.cremer@imb-mainz.de [Institute of Molecular Biology (IMB), Ackermannweg 4, 55128 Mainz (Germany); Institute for Pharmacy and Molecular Biotechnology (IPMB), University of Heidelberg, Im Neuenheimer Feld 364, D-69120 Heidelberg (Germany); Department of Physics, University of Mainz (JGU), Staudingerweg 7, 55128 Mainz (Germany)

    2016-05-01

    Higher order chromatin structure is not only required to compact and spatially arrange long chromatids within a nucleus, but have also important functional roles, including control of gene expression and DNA processing. However, studies of chromatin nanostructures cannot be performed using conventional widefield and confocal microscopy because of the limited optical resolution. Various methods of superresolution microscopy have been described to overcome this difficulty, like structured illumination and single molecule localization microscopy. We report here that the standard DNA dye Vybrant{sup ®} DyeCycle™ Violet can be used to provide single molecule localization microscopy (SMLM) images of DNA in nuclei of fixed mammalian cells. This SMLM method enabled optical isolation and localization of large numbers of DNA-bound molecules, usually in excess of 10{sup 6} signals in one cell nucleus. The technique yielded high-quality images of nuclear DNA density, revealing subdiffraction chromatin structures of the size in the order of 100 nm; the interchromatin compartment was visualized at unprecedented optical resolution. The approach offers several advantages over previously described high resolution DNA imaging methods, including high specificity, an ability to record images using a single wavelength excitation, and a higher density of single molecule signals than reported in previous SMLM studies. The method is compatible with DNA/multicolor SMLM imaging which employs simple staining methods suited also for conventional optical microscopy. - Highlights: • Super-resolution imaging of nuclear DNA with Vybrant Violet and blue excitation. • 90nm resolution images of DNA structures in optically thick eukaryotic nuclei. • Enhanced resolution confirms the existence of DNA-free regions inside the nucleus. • Optimized imaging conditions enable multicolor super-resolution imaging.

  2. Super-resolution mapping using multi-viewing CHRIS/PROBA data

    Science.gov (United States)

    Dwivedi, Manish; Kumar, Vinay

    2016-04-01

    High-spatial resolution Remote Sensing (RS) data provides detailed information which ensures high-definition visual image analysis of earth surface features. These data sets also support improved information extraction capabilities at a fine scale. In order to improve the spatial resolution of coarser resolution RS data, the Super Resolution Reconstruction (SRR) technique has become widely acknowledged which focused on multi-angular image sequences. In this study multi-angle CHRIS/PROBA data of Kutch area is used for SR image reconstruction to enhance the spatial resolution from 18 m to 6m in the hope to obtain a better land cover classification. Various SR approaches like Projection onto Convex Sets (POCS), Robust, Iterative Back Projection (IBP), Non-Uniform Interpolation and Structure-Adaptive Normalized Convolution (SANC) chosen for this study. Subjective assessment through visual interpretation shows substantial improvement in land cover details. Quantitative measures including peak signal to noise ratio and structural similarity are used for the evaluation of the image quality. It was observed that SANC SR technique using Vandewalle algorithm for the low resolution image registration outperformed the other techniques. After that SVM based classifier is used for the classification of SRR and data resampled to 6m spatial resolution using bi-cubic interpolation. A comparative analysis is carried out between classified data of bicubic interpolated and SR derived images of CHRIS/PROBA and SR derived classified data have shown a significant improvement of 10-12% in the overall accuracy. The results demonstrated that SR methods is able to improve spatial detail of multi-angle images as well as the classification accuracy.

  3. All-passive pixel super-resolution of time-stretch imaging

    Science.gov (United States)

    Chan, Antony C. S.; Ng, Ho-Cheung; Bogaraju, Sharat C. V.; So, Hayden K. H.; Lam, Edmund Y.; Tsia, Kevin K.

    2017-03-01

    Based on image encoding in a serial-temporal format, optical time-stretch imaging entails a stringent requirement of state-of-the-art fast data acquisition unit in order to preserve high image resolution at an ultrahigh frame rate — hampering the widespread utilities of such technology. Here, we propose a pixel super-resolution (pixel-SR) technique tailored for time-stretch imaging that preserves pixel resolution at a relaxed sampling rate. It harnesses the subpixel shifts between image frames inherently introduced by asynchronous digital sampling of the continuous time-stretch imaging process. Precise pixel registration is thus accomplished without any active opto-mechanical subpixel-shift control or other additional hardware. Here, we present the experimental pixel-SR image reconstruction pipeline that restores high-resolution time-stretch images of microparticles and biological cells (phytoplankton) at a relaxed sampling rate (≈2-5 GSa/s)—more than four times lower than the originally required readout rate (20 GSa/s) — is thus effective for high-throughput label-free, morphology-based cellular classification down to single-cell precision. Upon integration with the high-throughput image processing technology, this pixel-SR time-stretch imaging technique represents a cost-effective and practical solution for large scale cell-based phenotypic screening in biomedical diagnosis and machine vision for quality control in manufacturing.

  4. Super-Resolution for “Jilin-1” Satellite Video Imagery via a Convolutional Network

    Directory of Open Access Journals (Sweden)

    Aoran Xiao

    2018-04-01

    Full Text Available Super-resolution for satellite video attaches much significance to earth observation accuracy, and the special imaging and transmission conditions on the video satellite pose great challenges to this task. The existing deep convolutional neural-network-based methods require pre-processing or post-processing to be adapted to a high-resolution size or pixel format, leading to reduced performance and extra complexity. To this end, this paper proposes a five-layer end-to-end network structure without any pre-processing and post-processing, but imposes a reshape or deconvolution layer at the end of the network to retain the distribution of ground objects within the image. Meanwhile, we formulate a joint loss function by combining the output and high-dimensional features of a non-linear mapping network to precisely learn the desirable mapping relationship between low-resolution images and their high-resolution counterparts. Also, we use satellite video data itself as a training set, which favors consistency between training and testing images and promotes the method’s practicality. Experimental results on “Jilin-1” satellite video imagery show that this method demonstrates a superior performance in terms of both visual effects and measure metrics over competing methods.

  5. Super-Resolution for "Jilin-1" Satellite Video Imagery via a Convolutional Network.

    Science.gov (United States)

    Xiao, Aoran; Wang, Zhongyuan; Wang, Lei; Ren, Yexian

    2018-04-13

    Super-resolution for satellite video attaches much significance to earth observation accuracy, and the special imaging and transmission conditions on the video satellite pose great challenges to this task. The existing deep convolutional neural-network-based methods require pre-processing or post-processing to be adapted to a high-resolution size or pixel format, leading to reduced performance and extra complexity. To this end, this paper proposes a five-layer end-to-end network structure without any pre-processing and post-processing, but imposes a reshape or deconvolution layer at the end of the network to retain the distribution of ground objects within the image. Meanwhile, we formulate a joint loss function by combining the output and high-dimensional features of a non-linear mapping network to precisely learn the desirable mapping relationship between low-resolution images and their high-resolution counterparts. Also, we use satellite video data itself as a training set, which favors consistency between training and testing images and promotes the method's practicality. Experimental results on "Jilin-1" satellite video imagery show that this method demonstrates a superior performance in terms of both visual effects and measure metrics over competing methods.

  6. Internet-Enabled High-Resolution Brain Mapping and Virtual Microscopy

    OpenAIRE

    Mikula, Shawn; Trotts, Issac; Stone, James M.; Jones, Edward G.

    2007-01-01

    Virtual microscopy involves the conversion of histological sections mounted on glass microscope slides to high resolution digital images. Virtual microscopy offers several advantages over traditional microscopy, including remote viewing and data-sharing, annotation, and various forms of data-mining.

  7. Optimization of super-resolution processing using incomplete image sets in PET imaging.

    Science.gov (United States)

    Chang, Guoping; Pan, Tinsu; Clark, John W; Mawlawi, Osama R

    2008-12-01

    Super-resolution (SR) techniques are used in PET imaging to generate a high-resolution image by combining multiple low-resolution images that have been acquired from different points of view (POVs). The number of low-resolution images used defines the processing time and memory storage necessary to generate the SR image. In this paper, the authors propose two optimized SR implementations (ISR-1 and ISR-2) that require only a subset of the low-resolution images (two sides and diagonal of the image matrix, respectively), thereby reducing the overall processing time and memory storage. In an N x N matrix of low-resolution images, ISR-1 would be generated using images from the two sides of the N x N matrix, while ISR-2 would be generated from images across the diagonal of the image matrix. The objective of this paper is to investigate whether the two proposed SR methods can achieve similar performance in contrast and signal-to-noise ratio (SNR) as the SR image generated from a complete set of low-resolution images (CSR) using simulation and experimental studies. A simulation, a point source, and a NEMA/IEC phantom study were conducted for this investigation. In each study, 4 (2 x 2) or 16 (4 x 4) low-resolution images were reconstructed from the same acquired data set while shifting the reconstruction grid to generate images from different POVs. SR processing was then applied in each study to combine all as well as two different subsets of the low-resolution images to generate the CSR, ISR-1, and ISR-2 images, respectively. For reference purpose, a native reconstruction (NR) image using the same matrix size as the three SR images was also generated. The resultant images (CSR, ISR-1, ISR-2, and NR) were then analyzed using visual inspection, line profiles, SNR plots, and background noise spectra. The simulation study showed that the contrast and the SNR difference between the two ISR images and the CSR image were on average 0.4% and 0.3%, respectively. Line profiles of

  8. Optimized labeling of membrane proteins for applications to super-resolution imaging in confined cellular environments using monomeric streptavidin.

    Science.gov (United States)

    Chamma, Ingrid; Rossier, Olivier; Giannone, Grégory; Thoumine, Olivier; Sainlos, Matthieu

    2017-04-01

    Recent progress in super-resolution imaging (SRI) has created a strong need to improve protein labeling with probes of small size that minimize the target-to-label distance, increase labeling density, and efficiently penetrate thick biological tissues. This protocol describes a method for labeling genetically modified proteins incorporating a small biotin acceptor peptide with a 3-nm fluorescent probe, monomeric streptavidin. We show how to express, purify, and conjugate the probe to organic dyes with different fluorescent properties, and how to label selectively biotinylated membrane proteins for SRI techniques (point accumulation in nanoscale topography (PAINT), stimulated emission depletion (STED), stochastic optical reconstruction microscopy (STORM)). This method is complementary to the previously described anti-GFP-nanobody/SNAP-tag strategies, with the main advantage being that it requires only a short 15-amino-acid tag, and can thus be used with proteins resistant to fusion with large tags and for multicolor imaging. The protocol requires standard molecular biology/biochemistry equipment, making it easily accessible for laboratories with only basic skills in cell biology and biochemistry. The production/purification/conjugation steps take ∼5 d, and labeling takes a few minutes to an hour.

  9. Super-resolution by elliptical bubble formation with PtOx and AgInSbTe layers

    International Nuclear Information System (INIS)

    Kim, Jooho; Hwang, Inoh; Yoon, Duseop; Park, Insik; Shin, Dongho; Kikukawa, Takashi; Shima, Takayuki; Tominaga, Junji

    2003-01-01

    The recording and retrieval of signals below 100 nm mark length were attempted with elliptical bubble-type super-resolution technology with platinum oxide (PtO x ) and ductile AgInSbTe layers, using the same optical system as that of a digital versatile disk (a 635 nm wavelength red laser system). The carrier-to-noise ratio (CNR) of over 47 dB for 100 nm mark length signals (over 43 dB for 80 nm mark length signals) was obtained, which can be considered as a commercially acceptable level of CNR. The recording mechanism of the sample disk was shown through the transmission electron microscopy cross-section image observation to be by rigid elliptical bubble formation at the PtO x layer located between the AgInSbTe layers. The results of this report represent the potential for a much higher-density storage using the red laser system and a subterabyte optical storage using the blue laser system

  10. Deep Learning- and Transfer Learning-Based Super Resolution Reconstruction from Single Medical Image

    Directory of Open Access Journals (Sweden)

    YiNan Zhang

    2017-01-01

    Full Text Available Medical images play an important role in medical diagnosis and research. In this paper, a transfer learning- and deep learning-based super resolution reconstruction method is introduced. The proposed method contains one bicubic interpolation template layer and two convolutional layers. The bicubic interpolation template layer is prefixed by mathematics deduction, and two convolutional layers learn from training samples. For saving training medical images, a SIFT feature-based transfer learning method is proposed. Not only can medical images be used to train the proposed method, but also other types of images can be added into training dataset selectively. In empirical experiments, results of eight distinctive medical images show improvement of image quality and time reduction. Further, the proposed method also produces slightly sharper edges than other deep learning approaches in less time and it is projected that the hybrid architecture of prefixed template layer and unfixed hidden layers has potentials in other applications.

  11. From local pixel structure to global image super-resolution: a new face hallucination framework.

    Science.gov (United States)

    Hu, Yu; Lam, Kin-Man; Qiu, Guoping; Shen, Tingzhi

    2011-02-01

    We have developed a new face hallucination framework termed from local pixel structure to global image super-resolution (LPS-GIS). Based on the assumption that two similar face images should have similar local pixel structures, the new framework first uses the input low-resolution (LR) face image to search a face database for similar example high-resolution (HR) faces in order to learn the local pixel structures for the target HR face. It then uses the input LR face and the learned pixel structures as priors to estimate the target HR face. We present a three-step implementation procedure for the framework. Step 1 searches the database for K example faces that are the most similar to the input, and then warps the K example images to the input using optical flow. Step 2 uses the warped HR version of the K example faces to learn the local pixel structures for the target HR face. An effective method for learning local pixel structures from an individual face, and an adaptive procedure for fusing the local pixel structures of different example faces to reduce the influence of warping errors, have been developed. Step 3 estimates the target HR face by solving a constrained optimization problem by means of an iterative procedure. Experimental results show that our new method can provide good performances for face hallucination, both in terms of reconstruction error and visual quality; and that it is competitive with existing state-of-the-art methods.

  12. Robust Single Image Super-Resolution via Deep Networks With Sparse Prior.

    Science.gov (United States)

    Liu, Ding; Wang, Zhaowen; Wen, Bihan; Yang, Jianchao; Han, Wei; Huang, Thomas S

    2016-07-01

    Single image super-resolution (SR) is an ill-posed problem, which tries to recover a high-resolution image from its low-resolution observation. To regularize the solution of the problem, previous methods have focused on designing good priors for natural images, such as sparse representation, or directly learning the priors from a large data set with models, such as deep neural networks. In this paper, we argue that domain expertise from the conventional sparse coding model can be combined with the key ingredients of deep learning to achieve further improved results. We demonstrate that a sparse coding model particularly designed for SR can be incarnated as a neural network with the merit of end-to-end optimization over training data. The network has a cascaded structure, which boosts the SR performance for both fixed and incremental scaling factors. The proposed training and testing schemes can be extended for robust handling of images with additional degradation, such as noise and blurring. A subjective assessment is conducted and analyzed in order to thoroughly evaluate various SR techniques. Our proposed model is tested on a wide range of images, and it significantly outperforms the existing state-of-the-art methods for various scaling factors both quantitatively and perceptually.

  13. Single image super-resolution based on compressive sensing and improved TV minimization sparse recovery

    Science.gov (United States)

    Vishnukumar, S.; Wilscy, M.

    2017-12-01

    In this paper, we propose a single image Super-Resolution (SR) method based on Compressive Sensing (CS) and Improved Total Variation (TV) Minimization Sparse Recovery. In the CS framework, low-resolution (LR) image is treated as the compressed version of high-resolution (HR) image. Dictionary Training and Sparse Recovery are the two phases of the method. K-Singular Value Decomposition (K-SVD) method is used for dictionary training and the dictionary represents HR image patches in a sparse manner. Here, only the interpolated version of the LR image is used for training purpose and thereby the structural self similarity inherent in the LR image is exploited. In the sparse recovery phase the sparse representation coefficients with respect to the trained dictionary for LR image patches are derived using Improved TV Minimization method. HR image can be reconstructed by the linear combination of the dictionary and the sparse coefficients. The experimental results show that the proposed method gives better results quantitatively as well as qualitatively on both natural and remote sensing images. The reconstructed images have better visual quality since edges and other sharp details are preserved.

  14. Multisensor Super Resolution Using Directionally-Adaptive Regularization for UAV Images.

    Science.gov (United States)

    Kang, Wonseok; Yu, Soohwan; Ko, Seungyong; Paik, Joonki

    2015-05-22

    In various unmanned aerial vehicle (UAV) imaging applications, the multisensor super-resolution (SR) technique has become a chronic problem and attracted increasing attention. Multisensor SR algorithms utilize multispectral low-resolution (LR) images to make a higher resolution (HR) image to improve the performance of the UAV imaging system. The primary objective of the paper is to develop a multisensor SR method based on the existing multispectral imaging framework instead of using additional sensors. In order to restore image details without noise amplification or unnatural post-processing artifacts, this paper presents an improved regularized SR algorithm by combining the directionally-adaptive constraints and multiscale non-local means (NLM) filter. As a result, the proposed method can overcome the physical limitation of multispectral sensors by estimating the color HR image from a set of multispectral LR images using intensity-hue-saturation (IHS) image fusion. Experimental results show that the proposed method provides better SR results than existing state-of-the-art SR methods in the sense of objective measures.

  15. Super-resolution reconstruction of MR image with a novel residual learning network algorithm

    Science.gov (United States)

    Shi, Jun; Liu, Qingping; Wang, Chaofeng; Zhang, Qi; Ying, Shihui; Xu, Haoyu

    2018-04-01

    Spatial resolution is one of the key parameters of magnetic resonance imaging (MRI). The image super-resolution (SR) technique offers an alternative approach to improve the spatial resolution of MRI due to its simplicity. Convolutional neural networks (CNN)-based SR algorithms have achieved state-of-the-art performance, in which the global residual learning (GRL) strategy is now commonly used due to its effectiveness for learning image details for SR. However, the partial loss of image details usually happens in a very deep network due to the degradation problem. In this work, we propose a novel residual learning-based SR algorithm for MRI, which combines both multi-scale GRL and shallow network block-based local residual learning (LRL). The proposed LRL module works effectively in capturing high-frequency details by learning local residuals. One simulated MRI dataset and two real MRI datasets have been used to evaluate our algorithm. The experimental results show that the proposed SR algorithm achieves superior performance to all of the other compared CNN-based SR algorithms in this work.

  16. Single-Image Super-Resolution Based on Rational Fractal Interpolation.

    Science.gov (United States)

    Zhang, Yunfeng; Fan, Qinglan; Bao, Fangxun; Liu, Yifang; Zhang, Caiming

    2018-08-01

    This paper presents a novel single-image super-resolution (SR) procedure, which upscales a given low-resolution (LR) input image to a high-resolution image while preserving the textural and structural information. First, we construct a new type of bivariate rational fractal interpolation model and investigate its analytical properties. This model has different forms of expression with various values of the scaling factors and shape parameters; thus, it can be employed to better describe image features than current interpolation schemes. Furthermore, this model combines the advantages of rational interpolation and fractal interpolation, and its effectiveness is validated through theoretical analysis. Second, we develop a single-image SR algorithm based on the proposed model. The LR input image is divided into texture and non-texture regions, and then, the image is interpolated according to the characteristics of the local structure. Specifically, in the texture region, the scaling factor calculation is the critical step. We present a method to accurately calculate scaling factors based on local fractal analysis. Extensive experiments and comparisons with the other state-of-the-art methods show that our algorithm achieves competitive performance, with finer details and sharper edges.

  17. Super-resolution fluorescence imaging of membrane nanoscale architectures of hematopoietic stem cell homing and migration molecules

    KAUST Repository

    AbuZineh, Karmen

    2017-12-01

    Recent development of super-resolution (SR) fluorescence microscopy techniques has provided a new tool for direct visualization of subcellular structures and their dynamics in cells. The homing of Hematopoietic stem/progenitor cells (HSPCs) to bone marrow is a multistep process that is initiated by tethering of HSPCs to endothelium and mediated by spatiotemporally organised ligand-receptor interactions of selectins expressed on endothelial cells to their ligands expressed on HSPCs which occurs against the shear stress exerted by blood flow. Although molecules and biological processes involved in this multi-step cellular interaction have been studied extensively, molecular mechanisms of the homing, in particular the nanoscale spatiotemporal behaviour of ligand-receptor interactions and their role in the cellular interaction, remain elusive. Using our new method of microfluidics-based super-resolution fluorescence imaging platform we can now characterize the correlation between both nanoscale ligand-receptor interactions and tethering/rolling of cells under external shear stress. We found that cell rolling on E-selectin caused significant reorganization of the nanoscale clustering behavior of CD44 and CD43, from a patchy clusters of ~ 200 nm in size to an elongated network-like structures where for PSGL-1 the clustering size did not change significantly as it was 85 nm and after cell rolling the PSGL-1 aggregated to one side or even exhibited an increase in the footprint. Furthermore, I have established the use of 3D SR images that indicated that the patchy clusters of CD44 localize to protruding structures of the cell surface. On the other hand, a significant amount of the network-like elongated CD44 clusters observed after the rolling were located in the close proximity to the E-selectin surface. The effect of the nanoscale reorganization of the clusters on the HSPC rolling over selectins is still an open question at this stage. Nevertheless, my results further

  18. Synthetic biology's tall order: Reconstruction of 3D, super resolution images of single molecules in real-time

    CSIR Research Space (South Africa)

    Henriques, R

    2010-08-31

    Full Text Available -to-use reconstruction software coupled with image acquisition. Here, we present QuickPALM, an Image plugin, enabling real-time reconstruction of 3D super-resolution images during acquisition and drift correction. We illustrate its application by reconstructing Cy5...

  19. Ensemble variational Bayes tensor factorization for super resolution of CFRP debond detection

    Science.gov (United States)

    Lu, Peng; Gao, Bin; Feng, Qizhi; Yang, Yang; Woo, W. L.; Tian, Gui Yun

    2017-09-01

    The carbon fiber reinforced polymer (CFRP) is widely used in aircraft and wind turbine blades. The common type of CFRP defect is debond. Optical pulse thermographic nondestructive evaluation (OPTNDE) and relevant thermal feature extraction algorithms are generally used to detect the debond. However, the resolution of detection performance remain as challenges. In this paper, the ensemble variational Bayes tensor factorization has been proposed to conduct super resolution of the debond detection. The algorithm is based on the framework of variational Bayes tensor factorization and it constructs spatial-transient multi-layer mining structure which can significantly enhance the contrast ratio between the defective regions and sound regions. In order to quantitatively evaluate the results, the event based F-score is computed. The different information regions of the extracted thermal patterns are considered as different events and the purpose is to objectively evaluate the detectability for different algorithms. Experimental tests and comparative studies have been conducted to prove the efficacy of the proposed method.

  20. Super resolution reconstruction of μ-CT image of rock sample using neighbour embedding algorithm

    Science.gov (United States)

    Wang, Yuzhu; Rahman, Sheik S.; Arns, Christoph H.

    2018-03-01

    X-ray computed tomography (μ-CT) is considered to be the most effective way to obtain the inner structure of rock sample without destructions. However, its limited resolution hampers its ability to probe sub-micro structures which is critical for flow transportation of rock sample. In this study, we propose an innovative methodology to improve the resolution of μ-CT image using neighbour embedding algorithm where low frequency information is provided by μ-CT image itself while high frequency information is supplemented by high resolution scanning electron microscopy (SEM) image. In order to obtain prior for reconstruction, a large number of image patch pairs contain high- and low- image patches are extracted from the Gaussian image pyramid generated by SEM image. These image patch pairs contain abundant information about tomographic evolution of local porous structures under different resolution spaces. Relying on the assumption of self-similarity of porous structure, this prior information can be used to supervise the reconstruction of high resolution μ-CT image effectively. The experimental results show that the proposed method is able to achieve the state-of-the-art performance.

  1. Robust super-resolution by fusion of interpolated frames for color and grayscale images

    Directory of Open Access Journals (Sweden)

    Barry eKarch

    2015-04-01

    Full Text Available Multi-frame super-resolution (SR processing seeks to overcome undersampling issues that can lead to undesirable aliasing artifacts. The key to effective multi-frame SR is accurate subpixel inter-frame registration. This accurate registration is challenging when the motion does not obey a simple global translational model and may include local motion. SR processing is further complicated when the camera uses a division-of-focal-plane (DoFP sensor, such as the Bayer color filter array. Various aspects of these SR challenges have been previously investigated. Fast SR algorithms tend to have difficulty accommodating complex motion and DoFP sensors. Furthermore, methods that can tolerate these complexities tend to be iterative in nature and may not be amenable to real-time processing. In this paper, we present a new fast approach for performing SR in the presence of these challenging imaging conditions. We refer to the new approach as Fusion of Interpolated Frames (FIF SR. The FIF SR method decouples the demosaicing, interpolation, and restoration steps to simplify the algorithm. Frames are first individually demosaiced and interpolated to the desired resolution. Next, FIF uses a novel weighted sum of the interpolated frames to fuse them into an improved resolution estimate. Finally, restoration is applied to deconvolve the modeled system PSF. The proposed FIF approach has a lower computational complexity than most iterative methods, making it a candidate for real-time implementation. We provide a detailed description of the FIF SR method and show experimental results using synthetic and real datasets in both constrained and complex imaging scenarios. The experiments include airborne grayscale imagery and Bayer color array images with affine background motion plus local motion.

  2. Super-resolution imaging based on the temperature-dependent electron-phonon collision frequency effect of metal thin films

    Science.gov (United States)

    Ding, Chenliang; Wei, Jingsong; Xiao, Mufei

    2018-05-01

    We herein propose a far-field super-resolution imaging with metal thin films based on the temperature-dependent electron-phonon collision frequency effect. In the proposed method, neither fluorescence labeling nor any special properties are required for the samples. The 100 nm lands and 200 nm grooves on the Blu-ray disk substrates were clearly resolved and imaged through a laser scanning microscope of wavelength 405 nm. The spot size was approximately 0.80 μm , and the imaging resolution of 1/8 of the laser spot size was experimentally obtained. This work can be applied to the far-field super-resolution imaging of samples with neither fluorescence labeling nor any special properties.

  3. Image inpainting and super-resolution using non-local recursive deep convolutional network with skip connections

    Science.gov (United States)

    Liu, Miaofeng

    2017-07-01

    In recent years, deep convolutional neural networks come into use in image inpainting and super-resolution in many fields. Distinct to most of the former methods requiring to know beforehand the local information for corrupted pixels, we propose a 20-depth fully convolutional network to learn an end-to-end mapping a dataset of damaged/ground truth subimage pairs realizing non-local blind inpainting and super-resolution. As there often exist image with huge corruptions or inpainting on a low-resolution image that the existing approaches unable to perform well, we also share parameters in local area of layers to achieve spatial recursion and enlarge the receptive field. To avoid the difficulty of training this deep neural network, skip-connections between symmetric convolutional layers are designed. Experimental results shows that the proposed method outperforms state-of-the-art methods for diverse corrupting and low-resolution conditions, it works excellently when realizing super-resolution and image inpainting simultaneously

  4. Atomic Force Microscopy and Real Atomic Resolution. Simple Computer Simulations

    NARCIS (Netherlands)

    Koutsos, V.; Manias, E.; Brinke, G. ten; Hadziioannou, G.

    1994-01-01

    Using a simple computer simulation for AFM imaging in the contact mode, pictures with true and false atomic resolution are demonstrated. The surface probed consists of two f.c.c. (111) planes and an atomic vacancy is introduced in the upper layer. Changing the size of the effective tip and its

  5. High-resolution light microscopy using luminescent nanoparticles.

    Science.gov (United States)

    Ohulchanskyy, Tymish Y; Roy, Indrajit; Yong, Ken-Tye; Pudavar, Haridas E; Prasad, Paras N

    2010-01-01

    This review presents recent progress in the development of the luminescent nanoparticles for confocal and multiphoton microscopy. Four classes of nanomaterials are discussed: (1) silica-based nanoparticles doped with fluorescent molecules, (2) gold nanoparticles, (3) semiconductor nanocrystals (quantum dots/rods), and (4) nanophosphors. Special considerations are given to recently developed imaging nanoprobes, such as (1) organically modified silica (ORMOSIL) nanoparticles doped with two-photon absorbing fluorophores, which exhibit aggregation-enhanced fluorescence (AEF), and (2) nanophosphors (ceramic nanoparticles containing luminescent lanthanoid ions). Advantages and disadvantages of every class of nanomaterials and their specific applications are briefly discussed.

  6. Nobel Prize Recipient Eric Betzig Presents Lecture on Efforts to Improve High-Resolution Microscopy | Poster

    Science.gov (United States)

    Eric Betzig, Ph.D., a 2014 recipient of the Nobel Prize in Chemistry and a scientist at Janelia Research Campus (JRC), Howard Hughes Medical Institute, in Ashburn, Va., visited NCI at Frederick on Sept. 10 to present a Distinguished Scientist lecture and discuss the latest high-resolution microscopy techniques. Betzig co-invented photoactivation localization microscopy (PALM)

  7. Environmental high resolution electron microscopy and applications to chemical science

    OpenAIRE

    Boyes, Edward; Gai, Pratibha

    2017-01-01

    An environmental cell high resolution electron microscope (EHREM) has been developed for in situ studies of dynamic chemical reactions on the atomic scale. It allows access to metastable intermediate phases of catalysts and to sequences of reversible microstructural and chemical development associated with the activation, deactivation and poisoning of a catalyst. Materials transported through air can be restored or recreated and samples damaged, e.g. by dehydration, by the usual vacuum enviro...

  8. An Optofluidic Lens Array Microchip for High Resolution Stereo Microscopy

    Directory of Open Access Journals (Sweden)

    Mayurachat Ning Gulari

    2014-08-01

    Full Text Available We report the development of an add-on, chip-based, optical module—termed the Microfluidic-based Oil-immersion Lenses (μOIL chip—which transforms any stereo microscope into a high-resolution, large field of view imaging platform. The μOIL chip consists of an array of ball mini-lenses that are assembled onto a microfluidic silicon chip. The mini-lenses are made out of high refractive index material (sapphire and they are half immersed in oil. Those two key features enable submicron resolution and a maximum numerical aperture of ~1.2. The μOIL chip is reusable and easy to operate as it can be placed directly on top of any biological sample. It improves the resolution of a stereo microscope by an order of magnitude without compromising the field of view; therefore, we believe it could become a versatile tool for use in various research studies and clinical applications.

  9. Impact of detector-element active-area shape and fill factor on super-resolution

    Directory of Open Access Journals (Sweden)

    Russell Craig Hardie

    2015-05-01

    Full Text Available In many undersampled imaging systems, spatial integration from the individual detector elements is the dominant component of the system point spread function (PSF. Conventional focal plane arrays (FPAs utilize square detector elements with a nearly 100% fill factor, where fill factor is defined as the fraction of the detector element area that is active in light detection. A large fill factor is generally considered to be desirable because more photons are collected for a given pitch, and this leads to a higher signal-to-noise-ratio (SNR. However, the large active area works against super-resolution (SR image restoration by acting as an additional low pass filter in the overall PSF when modeled on the SR sampling grid. A high fill factor also tends to increase blurring from pixel cross-talk. In this paper, we study the impact of FPA detector-element shape and fill factor on SR. A detailed modulation transfer function analysis is provided along with a number of experimental results with both simulated data and real data acquired with a midwave infrared (MWIR imaging system. We demonstrate the potential advantage of low fill factor detector elements when combined with SR image restoration. Our results suggest that low fill factor circular detector elements may be the best choice. New video results are presented using robust adaptive Wiener filter SR processing applied to data from a commercial MWIR imaging system with both high and low detector element fill factors.

  10. Image super-resolution reconstruction based on regularization technique and guided filter

    Science.gov (United States)

    Huang, De-tian; Huang, Wei-qin; Gu, Pei-ting; Liu, Pei-zhong; Luo, Yan-min

    2017-06-01

    In order to improve the accuracy of sparse representation coefficients and the quality of reconstructed images, an improved image super-resolution algorithm based on sparse representation is presented. In the sparse coding stage, the autoregressive (AR) regularization and the non-local (NL) similarity regularization are introduced to improve the sparse coding objective function. A group of AR models which describe the image local structures are pre-learned from the training samples, and one or several suitable AR models can be adaptively selected for each image patch to regularize the solution space. Then, the image non-local redundancy is obtained by the NL similarity regularization to preserve edges. In the process of computing the sparse representation coefficients, the feature-sign search algorithm is utilized instead of the conventional orthogonal matching pursuit algorithm to improve the accuracy of the sparse coefficients. To restore image details further, a global error compensation model based on weighted guided filter is proposed to realize error compensation for the reconstructed images. Experimental results demonstrate that compared with Bicubic, L1SR, SISR, GR, ANR, NE + LS, NE + NNLS, NE + LLE and A + (16 atoms) methods, the proposed approach has remarkable improvement in peak signal-to-noise ratio, structural similarity and subjective visual perception.

  11. Cell-specific STORM super-resolution imaging reveals nanoscale organization of cannabinoid signaling.

    Science.gov (United States)

    Dudok, Barna; Barna, László; Ledri, Marco; Szabó, Szilárd I; Szabadits, Eszter; Pintér, Balázs; Woodhams, Stephen G; Henstridge, Christopher M; Balla, Gyula Y; Nyilas, Rita; Varga, Csaba; Lee, Sang-Hun; Matolcsi, Máté; Cervenak, Judit; Kacskovics, Imre; Watanabe, Masahiko; Sagheddu, Claudia; Melis, Miriam; Pistis, Marco; Soltesz, Ivan; Katona, István

    2015-01-01

    A major challenge in neuroscience is to determine the nanoscale position and quantity of signaling molecules in a cell type- and subcellular compartment-specific manner. We developed a new approach to this problem by combining cell-specific physiological and anatomical characterization with super-resolution imaging and studied the molecular and structural parameters shaping the physiological properties of synaptic endocannabinoid signaling in the mouse hippocampus. We found that axon terminals of perisomatically projecting GABAergic interneurons possessed increased CB1 receptor number, active-zone complexity and receptor/effector ratio compared with dendritically projecting interneurons, consistent with higher efficiency of cannabinoid signaling at somatic versus dendritic synapses. Furthermore, chronic Δ(9)-tetrahydrocannabinol administration, which reduces cannabinoid efficacy on GABA release, evoked marked CB1 downregulation in a dose-dependent manner. Full receptor recovery required several weeks after the cessation of Δ(9)-tetrahydrocannabinol treatment. These findings indicate that cell type-specific nanoscale analysis of endogenous protein distribution is possible in brain circuits and identify previously unknown molecular properties controlling endocannabinoid signaling and cannabis-induced cognitive dysfunction.

  12. 3D Super-Resolution Motion-Corrected MRI: Validation of Fetal Posterior Fossa Measurements.

    Science.gov (United States)

    Pier, Danielle B; Gholipour, Ali; Afacan, Onur; Velasco-Annis, Clemente; Clancy, Sean; Kapur, Kush; Estroff, Judy A; Warfield, Simon K

    2016-09-01

    Current diagnosis of fetal posterior fossa anomalies by sonography and conventional MRI is limited by fetal position, motion, and by two-dimensional (2D), rather than three-dimensional (3D), representation. In this study, we aimed to validate the use of a novel magnetic resonance imaging (MRI) technique, 3D super-resolution motion-corrected MRI, to image the fetal posterior fossa. From a database of pregnant women who received fetal MRIs at our institution, images of 49 normal fetal brains were reconstructed. Six measurements of the cerebellum, vermis, and pons were obtained for all cases on 2D conventional and 3D reconstructed MRI, and the agreement between the two methods was determined using concordance correlation coefficients. Concordance of axial and coronal measurements of the transcerebellar diameter was also assessed within each method. Between the two methods, the concordance of measurements was high for all six structures (P fetal motion and orthogonal slice acquisition. This technique will facilitate further study of fetal abnormalities of the posterior fossa. Copyright © 2016 by the American Society of Neuroimaging.

  13. All-optical control and super-resolution imaging of quantum emitters in layered materials.

    Science.gov (United States)

    Kianinia, Mehran; Bradac, Carlo; Sontheimer, Bernd; Wang, Fan; Tran, Toan Trong; Nguyen, Minh; Kim, Sejeong; Xu, Zai-Quan; Jin, Dayong; Schell, Andreas W; Lobo, Charlene J; Aharonovich, Igor; Toth, Milos

    2018-02-28

    Layered van der Waals materials are emerging as compelling two-dimensional platforms for nanophotonics, polaritonics, valleytronics and spintronics, and have the potential to transform applications in sensing, imaging and quantum information processing. Among these, hexagonal boron nitride (hBN) is known to host ultra-bright, room-temperature quantum emitters, whose nature is yet to be fully understood. Here we present a set of measurements that give unique insight into the photophysical properties and level structure of hBN quantum emitters. Specifically, we report the existence of a class of hBN quantum emitters with a fast-decaying intermediate and a long-lived metastable state accessible from the first excited electronic state. Furthermore, by means of a two-laser repumping scheme, we show an enhanced photoluminescence and emission intensity, which can be utilized to realize a new modality of far-field super-resolution imaging. Our findings expand current understanding of quantum emitters in hBN and show new potential ways of harnessing their nonlinear optical properties in sub-diffraction nanoscopy.

  14. Improved Wallis Dodging Algorithm for Large-Scale Super-Resolution Reconstruction Remote Sensing Images

    Directory of Open Access Journals (Sweden)

    Chong Fan

    2017-03-01

    Full Text Available A sub-block algorithm is usually applied in the super-resolution (SR reconstruction of images because of limitations in computer memory. However, the sub-block SR images can hardly achieve a seamless image mosaicking because of the uneven distribution of brightness and contrast among these sub-blocks. An effectively improved weighted Wallis dodging algorithm is proposed, aiming at the characteristic that SR reconstructed images are gray images with the same size and overlapping region. This algorithm can achieve consistency of image brightness and contrast. Meanwhile, a weighted adjustment sequence is presented to avoid the spatial propagation and accumulation of errors and the loss of image information caused by excessive computation. A seam line elimination method can share the partial dislocation in the seam line to the entire overlapping region with a smooth transition effect. Subsequently, the improved method is employed to remove the uneven illumination for 900 SR reconstructed images of ZY-3. Then, the overlapping image mosaic method is adopted to accomplish a seamless image mosaic based on the optimal seam line.

  15. Newmark-Beta-FDTD method for super-resolution analysis of time reversal waves

    Science.gov (United States)

    Shi, Sheng-Bing; Shao, Wei; Ma, Jing; Jin, Congjun; Wang, Xiao-Hua

    2017-09-01

    In this work, a new unconditionally stable finite-difference time-domain (FDTD) method with the split-field perfectly matched layer (PML) is proposed for the analysis of time reversal (TR) waves. The proposed method is very suitable for multiscale problems involving microstructures. The spatial and temporal derivatives in this method are discretized by the central difference technique and Newmark-Beta algorithm, respectively, and the derivation results in the calculation of a banded-sparse matrix equation. Since the coefficient matrix keeps unchanged during the whole simulation process, the lower-upper (LU) decomposition of the matrix needs to be performed only once at the beginning of the calculation. Moreover, the reverse Cuthill-Mckee (RCM) technique, an effective preprocessing technique in bandwidth compression of sparse matrices, is used to improve computational efficiency. The super-resolution focusing of TR wave propagation in two- and three-dimensional spaces is included to validate the accuracy and efficiency of the proposed method.

  16. Extending the Capture Volume of an Iris Recognition System Using Wavefront Coding and Super-Resolution.

    Science.gov (United States)

    Hsieh, Sheng-Hsun; Li, Yung-Hui; Tien, Chung-Hao; Chang, Chin-Chen

    2016-12-01

    Iris recognition has gained increasing popularity over the last few decades; however, the stand-off distance in a conventional iris recognition system is too short, which limits its application. In this paper, we propose a novel hardware-software hybrid method to increase the stand-off distance in an iris recognition system. When designing the system hardware, we use an optimized wavefront coding technique to extend the depth of field. To compensate for the blurring of the image caused by wavefront coding, on the software side, the proposed system uses a local patch-based super-resolution method to restore the blurred image to its clear version. The collaborative effect of the new hardware design and software post-processing showed great potential in our experiment. The experimental results showed that such improvement cannot be achieved by using a hardware-or software-only design. The proposed system can increase the capture volume of a conventional iris recognition system by three times and maintain the system's high recognition rate.

  17. A Search for Water in a Super-Earth Atmosphere: High-resolution Optical Spectroscopy of 55Cancri e

    Energy Technology Data Exchange (ETDEWEB)

    Esteves, Lisa J. [Astronomy and Astrophysics, University of Toronto, 50 St. George Street, Toronto, Ontario M5S 3H4 (Canada); De Mooij, Ernst J. W.; Watson, Chris [Astrophysics Research Centre, School of Mathematics and Physics, Queens University, Belfast (United Kingdom); Jayawardhana, Ray [Physics and Astronomy, York University, Toronto, Ontario L3T 3R1 (Canada); De Kok, Remco, E-mail: esteves@astro.utoronto.ca, E-mail: ernst.demooij@dcu.ie, E-mail: c.a.watson@qub.ac.uk, E-mail: rayjay@yorku.ca, E-mail: r.j.de.kok@sron.nl [Leiden Observatory, Leiden University, Postbus 9513, 2300 RA, Leiden (Netherlands)

    2017-06-01

    We present the analysis of high-resolution optical spectra of four transits of 55Cnc e, a low-density super-Earth that orbits a nearby Sun-like star in under 18 hr. The inferred bulk density of the planet implies a substantial envelope, which, according to mass–radius relationships, could be either a low-mass extended or a high-mass compact atmosphere. Our observations investigate the latter scenario, with water as the dominant species. We take advantage of the Doppler cross-correlation technique, high-spectral resolution, and the large wavelength coverage of our observations to search for the signature of thousands of optical water absorption lines. Using our observations with HDS on the Subaru telescope and ESPaDOnS on the Canada–France–Hawaii Telescope, we are able to place a 3 σ lower limit of 10 g mol{sup −1} on the mean-molecular weight of 55Cnc e’s water-rich (volume mixing ratio >10%), optically thin atmosphere, which corresponds to an atmospheric scale-height of ∼80 km. Our study marks the first high-spectral resolution search for water in a super-Earth atmosphere, and demonstrates that it is possible to recover known water-vapor absorption signals in a nearby super-Earth atmosphere, using high-resolution transit spectroscopy with current ground-based instruments.

  18. Imaging three-dimensional surface objects with submolecular resolution by atomic force microscopy

    Czech Academy of Sciences Publication Activity Database

    Moreno, C.; Stetsovych, Oleksandr; Shimizu, T.K.; Custance, O.

    2015-01-01

    Roč. 15, č. 4 (2015), s. 2257-2262 ISSN 1530-6984 Institutional support: RVO:68378271 Keywords : noncontact atomic force microscopy (NC- AFM ) * submolecular resolution * three-dimensional dynamic force spectroscopy * high-resolution imaging Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 13.779, year: 2015

  19. Super-TIGER-2: A Very-Large-Area, High-Resolution Trans-Iron Cosmic Ray Investigation

    Science.gov (United States)

    Binns, Walter

    This is the lead proposal of a multi-institution proposal. We propose to continue the highly successful Super-TIGER (Super Trans-Iron Galactic Element Recorder) program and to extend its scientific reach. Super-TIGER is a large-area instrument designed to make precision measurements of the elemental composition of ultra-heavy cosmic rays (UHCR) with atomic number Z greater than or equal to 30. The principal objective of the first phase of the Super- TIGER program was to measure the abundances of nuclei with 30 less than or equal to Z less than or equal to 42 with clear individual element resolution and high statistical precision. A secondary objective was to accurately measure the energy spectra of the more abundant light elements with 12 less than or equal to Z less than or equal to 28. Super-TIGER-1 was flown during the 2012-2013 Austral Summer, returning data on over 50 million cosmic ray (CR) nuclei in 55 days at float. The excellent data from this flight should enable us to achieve the initial goals of the program, and the high performance of the instrument makes it possible to expand our primary objective for further flights to include heavier UHCR. This is a 1-year proposal with two objectives: First to complete analysis of the data from the Super-TIGER-1 flight, and second to begin preparations to extend UHCR measurements with individual element resolution through barium (Z=56) and to greatly increase the number of Z greater than or equal to 30 nuclei measured. The abundance measurements provide sensitive tests and clarification of the OB-association model of galactic cosmic-ray origins, and will test models for atomic processes by which nuclei are selected for acceleration to cosmic ray energies. Additionally, measurements of individual element abundances from Z=40 to 56 will enable us to determine the extent of r-process enhancement since Zr (Z=40), Sn (Z=50) and Ba (Z=56) are predominately s-process and Ru (Z=44), Pd (Z=46), Te (Z=52) and Xe (Z=54)are

  20. Observation of Eye Pattern on Super-Resolution Near-Field Structure Disk with Write-Strategy Technique

    Science.gov (United States)

    Fuji, Hiroshi; Kikukawa, Takashi; Tominaga, Junji

    2004-07-01

    Pit-edge recording at a density of 150 nm pits and spaces is carried out on a super-resolution near-field structure (super-RENS) disk with a platinum oxide layer. Pits are recorded and read using a 635-nm-wavelength laser and an objective lens with a 0.6 numerical aperture. We arrange laser pulses to correctly record the pits on the disk by a write-strategy technique. The laser-pulse figure includes a unit time of 0.25 T and intensities of Pw1, Pw2 and Pw3. After recording pits of various lengths, the observation of an eye pattern is achieved despite a pit smaller than the resolution limit. Furthermore, the eye pattern maintains its shape even though other pits fill the adjacent tracks at a track density of 600 nm. The disk can be used as a pit-edge recording system through a write-strategy technique.

  1. High-resolution multiphoton microscopy with a low-power continuous wave laser pump.

    Science.gov (United States)

    Chen, Xiang-Dong; Li, Shen; Du, Bo; Dong, Yang; Wang, Ze-Hao; Guo, Guang-Can; Sun, Fang-Wen

    2018-02-15

    Multiphoton microscopy (MPM) has been widely used for three-dimensional biological imaging. Here, based on the photon-induced charge state conversion process, we demonstrated a low-power high-resolution MPM with a nitrogen vacancy (NV) center in diamond. Continuous wave green and orange lasers were used to pump and detect the two-photon charge state conversion, respectively. The power of the laser for multiphoton excitation was 40 μW. Both the axial and lateral resolutions were improved approximately 1.5 times compared with confocal microscopy. The results can be used to improve the resolution of the NV center-based quantum sensing and biological imaging.

  2. High-resolution optical microscopy of carbon and graphite

    International Nuclear Information System (INIS)

    Cook, W.H.; Allen, M.D.; Leslie, B.C.; Gray, R.J.

    1975-01-01

    The ceramographic preparation of carbonaceous materials varying in crystalline quality, amorphous carbon to well crystallized graphite, is described. In a two-step process, using alumina and diamond polishing compounds, one can prepare more samples, obtain a substantial saving in man hours, avoid rounding material around pores, and obtain flatter surfaces than were obtainable with earlier, conventional methods. Improved resolution of microstructural details is achieved without impregnation with epoxy resins or other materials to support the porous structures. Use of rotatable, half-wave retardation (sensitive tint) enhances the microstructural definition in both color and black and white. These innovations were extensively used as part of the examination of nuclear grades of graphite before and after exposure to fast neutrons at temperatures from 650 to 1100 0 C; typical examples are discussed. (auth)

  3. Brain Atlas Fusion from High-Thickness Diagnostic Magnetic Resonance Images by Learning-Based Super-Resolution.

    Science.gov (United States)

    Zhang, Jinpeng; Zhang, Lichi; Xiang, Lei; Shao, Yeqin; Wu, Guorong; Zhou, Xiaodong; Shen, Dinggang; Wang, Qian

    2017-03-01

    It is fundamentally important to fuse the brain atlas from magnetic resonance (MR) images for many imaging-based studies. Most existing works focus on fusing the atlases from high-quality MR images. However, for low-quality diagnostic images (i.e., with high inter-slice thickness), the problem of atlas fusion has not been addressed yet. In this paper, we intend to fuse the brain atlas from the high-thickness diagnostic MR images that are prevalent for clinical routines. The main idea of our works is to extend the conventional groupwise registration by incorporating a novel super-resolution strategy. The contribution of the proposed super-resolution framework is two-fold. First, each high-thickness subject image is reconstructed to be isotropic by the patch-based sparsity learning. Then, the reconstructed isotropic image is enhanced for better quality through the random-forest-based regression model. In this way, the images obtained by the super-resolution strategy can be fused together by applying the groupwise registration method to construct the required atlas. Our experiments have shown that the proposed framework can effectively solve the problem of atlas fusion from the low-quality brain MR images.

  4. Super-resolution fluorescence imaging of nanoimprinted polymer patterns by selective fluorophore adsorption combined with redox switching

    Directory of Open Access Journals (Sweden)

    Yu Yabiku

    2013-10-01

    Full Text Available We applied a super-resolution fluorescence imaging based on selective adsorption and redox switching of the fluorescent dye molecules for studying polymer nanostructures. We demonstrate that nano-scale structures of polymer thin films can be visualized with the image resolution better than 80 nm. The method was applied to image 100 nm-wide polymer nanopatterns fabricated by thermal nanoimprinting. The results point to the applicability of the method for evaluating residual polymer thin films and dewetting defect of the polymer resist patterns which are important for the quality control of the fine nanoimprinted patterns.

  5. Triple-color super-resolution imaging of live cells: resolving submicroscopic receptor organization in the plasma membrane.

    Science.gov (United States)

    Wilmes, Stephan; Staufenbiel, Markus; Lisse, Domenik; Richter, Christian P; Beutel, Oliver; Busch, Karin B; Hess, Samuel T; Piehler, Jacob

    2012-05-14

    In living color: efficient intracellular covalent labeling of proteins with a photoswitchable dye using the HaloTag for dSTORM super-resolution imaging in live cells is described. The dynamics of cellular nanostructures at the plasma membrane were monitored with a time resolution of a few seconds. In combination with dual-color FPALM imaging, submicroscopic receptor organization within the context of the membrane skeleton was resolved. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Super-resolution fluorescence imaging of nanoimprinted polymer patterns by selective fluorophore adsorption combined with redox switching

    KAUST Repository

    Yabiku, Y.

    2013-10-22

    We applied a super-resolution fluorescence imaging based on selective adsorption and redox switching of the fluorescent dye molecules for studying polymer nanostructures. We demonstrate that nano-scale structures of polymer thin films can be visualized with the image resolution better than 80 nm. The method was applied to image 100 nm-wide polymer nanopatterns fabricated by thermal nanoimprinting. The results point to the applicability of the method for evaluating residual polymer thin films and dewetting defect of the polymer resist patterns which are important for the quality control of the fine nanoimprinted patterns. 2013 Author(s).

  7. Transceiver Design for CMUT-Based Super-Resolution Ultrasound Imaging.

    Science.gov (United States)

    Behnamfar, Parisa; Molavi, Reza; Mirabbasi, Shahriar

    2016-04-01

    A recently introduced structure for the capacitive micromachined ultrasonic transducers (CMUTs) has focused on the applications of the asymmetric mode of vibration and has shown promising results in construction of super-resolution ultrasound images. This paper presents the first implementation and experimental results of a transceiver circuit to interface such CMUT structures. The multiple input/multiple output receiver in this work supports both fundamental and asymmetric modes of operation and includes transimpedance amplifiers and low-power variable-gain stages. These circuit blocks are designed considering the trade-offs between gain, input impedance, noise, linearity and power consumption. The high-voltage transmitter can generate pulse voltages up to 60 V while occupying a considerably small area. The overall circuit is designed and laid out in a 0.35 μm CMOS process and a four-channel transceiver occupies 0.86 × 0.38 mm(2). The prototype chip is characterized in both electrical and mechanical domains. Measurement results show that each receiver channel has a nominal gain of 110 dBΩ with a 3 dB bandwidth of 9 MHz while consuming 1.02 mW from a 3.3 V supply. The receiver is also highly linear, with 1 dB compression point of minimum 1.05 V which is considerably higher than the previously reported designs. The transmitter consumes 98.1 mW from a 30 V supply while generating 1.38 MHz, 30 V pulses. The CMOS-CMUT system is tested in the transmit mode and shows full functionality in air medium.

  8. Super-Resolution Community Detection for Layer-Aggregated Multilayer Networks

    Directory of Open Access Journals (Sweden)

    Dane Taylor

    2017-09-01

    Full Text Available Applied network science often involves preprocessing network data before applying a network-analysis method, and there is typically a theoretical disconnect between these steps. For example, it is common to aggregate time-varying network data into windows prior to analysis, and the trade-offs of this preprocessing are not well understood. Focusing on the problem of detecting small communities in multilayer networks, we study the effects of layer aggregation by developing random-matrix theory for modularity matrices associated with layer-aggregated networks with N nodes and L layers, which are drawn from an ensemble of Erdős–Rényi networks with communities planted in subsets of layers. We study phase transitions in which eigenvectors localize onto communities (allowing their detection and which occur for a given community provided its size surpasses a detectability limit K^{*}. When layers are aggregated via a summation, we obtain K^{*}∝O(sqrt[NL]/T, where T is the number of layers across which the community persists. Interestingly, if T is allowed to vary with L, then summation-based layer aggregation enhances small-community detection even if the community persists across a vanishing fraction of layers, provided that T/L decays more slowly than O(L^{-1/2}. Moreover, we find that thresholding the summation can, in some cases, cause K^{*} to decay exponentially, decreasing by orders of magnitude in a phenomenon we call super-resolution community detection. In other words, layer aggregation with thresholding is a nonlinear data filter enabling detection of communities that are otherwise too small to detect. Importantly, different thresholds generally enhance the detectability of communities having different properties, illustrating that community detection can be obscured if one analyzes network data using a single threshold.

  9. Super-Resolution Community Detection for Layer-Aggregated Multilayer Networks.

    Science.gov (United States)

    Taylor, Dane; Caceres, Rajmonda S; Mucha, Peter J

    2017-01-01

    Applied network science often involves preprocessing network data before applying a network-analysis method, and there is typically a theoretical disconnect between these steps. For example, it is common to aggregate time-varying network data into windows prior to analysis, and the trade-offs of this preprocessing are not well understood. Focusing on the problem of detecting small communities in multilayer networks, we study the effects of layer aggregation by developing random-matrix theory for modularity matrices associated with layer-aggregated networks with N nodes and L layers, which are drawn from an ensemble of Erdős-Rényi networks with communities planted in subsets of layers. We study phase transitions in which eigenvectors localize onto communities (allowing their detection) and which occur for a given community provided its size surpasses a detectability limit K * . When layers are aggregated via a summation, we obtain [Formula: see text], where T is the number of layers across which the community persists. Interestingly, if T is allowed to vary with L , then summation-based layer aggregation enhances small-community detection even if the community persists across a vanishing fraction of layers, provided that T/L decays more slowly than ( L -1/2 ). Moreover, we find that thresholding the summation can, in some cases, cause K * to decay exponentially, decreasing by orders of magnitude in a phenomenon we call super-resolution community detection. In other words, layer aggregation with thresholding is a nonlinear data filter enabling detection of communities that are otherwise too small to detect. Importantly, different thresholds generally enhance the detectability of communities having different properties, illustrating that community detection can be obscured if one analyzes network data using a single threshold.

  10. Quantum phase amplification for temporal pulse shaping and super-resolution in remote sensing

    Science.gov (United States)

    Yin, Yanchun

    QPA in the spatial domain has also been studied as a method to enhance the spatial resolution of imaging systems. A detailed model has been developed for achieving both super-resolution and detection of phase-amplified light. The imaging resolution problem considered here is treated as a binary hypotheses testing problem. Resolution enhancement is achieved by magnification of the angular separation of two targets in the sub-Rayleigh regime. The detection model includes optimization of detector segmentation, detector noise, and detection in both the spatial and the spatial frequency domain, which provide strategies for the optimization of the signal-to-noise ratio that take advantage of both the change of the field distribution and the change of energy of the signal in the QPA process. Proof-of-principle experiments have been conducted in the spatial domain. For the first time, beam angular amplification has been demonstrated, and the experimental result is in good agreement with simulations. The experimental demonstration has been achieved by observing the correlation of amplitude and angular phase in the phase-sensitive three-wave mixing process using ultrashort laser pulses and utilizing a type I three-wave mixing process. Several diagnostics have been developed and employed in the experimental measurements, including the near-field diagnostic, the far-field diagnostic, and the interferometry diagnostic. They have all been used to confirm the existence and study the properties of the QPA process on a shot-to-shot basis. Specifically, amplitude was measured in the near-field diagnostic, while the angular phase was indirectly measured in the far-field diagnostic by determining the position of the beam centroid. Interferometric measurements have been found to be of insufficient accuracy for this measurement in the way they were implemented. The demonstration of beam angular amplification by use of QPA lays the foundation for future integrated demonstration of imaging

  11. Sparse coded image super-resolution using K-SVD trained dictionary based on regularized orthogonal matching pursuit.

    Science.gov (United States)

    Sajjad, Muhammad; Mehmood, Irfan; Baik, Sung Wook

    2015-01-01

    Image super-resolution (SR) plays a vital role in medical imaging that allows a more efficient and effective diagnosis process. Usually, diagnosing is difficult and inaccurate from low-resolution (LR) and noisy images. Resolution enhancement through conventional interpolation methods strongly affects the precision of consequent processing steps, such as segmentation and registration. Therefore, we propose an efficient sparse coded image SR reconstruction technique using a trained dictionary. We apply a simple and efficient regularized version of orthogonal matching pursuit (ROMP) to seek the coefficients of sparse representation. ROMP has the transparency and greediness of OMP and the robustness of the L1-minization that enhance the dictionary learning process to capture feature descriptors such as oriented edges and contours from complex images like brain MRIs. The sparse coding part of the K-SVD dictionary training procedure is modified by substituting OMP with ROMP. The dictionary update stage allows simultaneously updating an arbitrary number of atoms and vectors of sparse coefficients. In SR reconstruction, ROMP is used to determine the vector of sparse coefficients for the underlying patch. The recovered representations are then applied to the trained dictionary, and finally, an optimization leads to high-resolution output of high-quality. Experimental results demonstrate that the super-resolution reconstruction quality of the proposed scheme is comparatively better than other state-of-the-art schemes.

  12. Super-Resolution Molecular and Functional Imaging of Nanoscale Architectures in Life and Materials Science

    KAUST Repository

    Habuchi, Satoshi

    2014-01-01

    fluorescence microscopy techniques along with the latest developments of fluorophores and labeling for the SR microscopy. I discuss the applications of SR microscopy in the fields of life science and materials science with a special emphasis on quantitative

  13. A novel method for enhancing the lateral resolution and image SNR in confocal microscopy

    Science.gov (United States)

    Chen, Youhua; Zhu, Dazhao; Fang, Yue; Kuang, Cuifang; Liu, Xu

    2017-12-01

    There is always a tradeoff between the resolution and the signal-to-noise ratio (SNR) in confocal microscopy. In particular, the pinhole size is very important for maintaining a balance between them. In this paper, we propose a method for improving the lateral resolution and image SNR in confocal microscopy without making any changes to the hardware. By using the fluorescence emission difference (FED) approach, we divide the images acquired by different pinhole sizes into one image acquired by the central pinhole and several images acquired by ring-shaped pinholes. Then, they are added together with the deconvolution method. Simulation and experimental results for fluorescent particles and cells show that our method can achieve a far better resolution than a large pinhole and a higher SNR than a small pinhole. Moreover, our method can improve the performance of classic confocal laser scanning microscopy (CLSM) to a certain extent, especially CLSM with a continuously variable pinhole.

  14. Instrumentation at the National Center for Electron Microscopy: the Atomic Resolution Microscope

    International Nuclear Information System (INIS)

    Gronsky, R.; Thomas, G.

    1983-01-01

    The Atomic Resolution Microscope (ARM) is one of two unique high voltage electron microscopes at the Lawrence Berkeley Laboratory's National Center for Electron Microscopy (NCEM). The latest results from this new instrument which was manufactured by JEOL, Ltd. to the performance specifications of the NCEM, delivered in January of 1983, and soon to be open to access by the entire microscopy community are given. Details of its history and development are given and its performance specifications are reviewed

  15. Enhancement of fluorescence confocal scanning microscopy lateral resolution by use of structured illumination

    International Nuclear Information System (INIS)

    Kim, Taejoong; Gweon, DaeGab; Lee, Jun-Hee

    2009-01-01

    Confocal microscopy is an optical imaging technique used to reconstruct three-dimensional images without physical sectioning. As with other optical microscopes, the lateral resolution of the confocal microscope cannot surpass the diffraction limit. This paper presents a novel imaging system, structured illumination confocal scanning microscopy (SICSM), that uses structured illumination to improve the lateral resolution of the confocal microscope. The SICSM can easily be implemented by introducing a structured illumination generating optics to conventional line-scanning fluorescence confocal microscopy. In this paper, we report our analysis of the lateral and axial resolutions of the SICSM by use of mathematical imaging theory. Numerical simulation results show that the lateral resolution of the SICSM is 1.43-fold better than that of the confocal microscope. In the axial direction, however, the resolution of the SICSM is ∼15% poorer than that of the confocal microscope. This deterioration arises because of a decrease in the axial cut-off frequency caused by the process of generating structured illumination. We propose the use of imaging conditions under which a compromise between the axial and lateral resolutions is chosen. Finally, we show simulated images of diversely shaped test objects to demonstrate the lateral and axial resolution performance of the SICSM

  16. Atomic resolution scanning tunneling microscopy in a cryogen free dilution refrigerator at 15 mK

    International Nuclear Information System (INIS)

    Haan, A. M. J. den; Wijts, G. H. C. J.; Galli, F.; Oosterkamp, T. H.; Usenko, O.; Baarle, G. J. C. van; Zalm, D. J. van der

    2014-01-01

    Pulse tube refrigerators are becoming more common, because they are cost efficient and demand less handling than conventional (wet) refrigerators. However, a downside of a pulse tube system is the vibration level at the cold-head, which is in most designs several micrometers. We implemented vibration isolation techniques which significantly reduced vibration levels at the experiment. These optimizations were necessary for the vibration sensitive magnetic resonance force microscopy experiments at milli-kelvin temperatures for which the cryostat is intended. With these modifications we show atomic resolution scanning tunneling microscopy on graphite. This is promising for scanning probe microscopy applications at very low temperatures

  17. Super-resolution microscopy reveals a preformed NEMO lattice structure that is collapsed in incontinentia pigmenti

    CSIR Research Space (South Africa)

    Scholefield, Janine

    2016-09-01

    Full Text Available Boucharlat4, Emmanuel Laplantine5, Asma Smahi6, Alain Israe¨l5, Fabrice Agou4 & Musa M. Mhlanga1,2,7 The NF-kB pathway has critical roles in cancer, immunity and inflammatory responses. Understanding the mechanism(s) by which mutations in genes involved...

  18. Diverse Protocols for Correlative Super-Resolution Fluorescence Imaging and Electron Microscopy of Cells and Tissue

    Science.gov (United States)

    2016-05-25

    polymerization36. Finally, we mutated surface residues on mEos2 to remove nucleophilic groups, which are involved in cross-linking with aldehydes and...Sites of Clathrin-Mediated Endocytosis. Current Biology 21, 1167– 1175 (2011). 49. Caplan, J. et al. Correlative Protein Localization in Yeast . (2013

  19. Spatio-Temporal Super-Resolution Reconstruction of Remote-Sensing Images Based on Adaptive Multi-Scale Detail Enhancement.

    Science.gov (United States)

    Zhu, Hong; Tang, Xinming; Xie, Junfeng; Song, Weidong; Mo, Fan; Gao, Xiaoming

    2018-02-07

    There are many problems in existing reconstruction-based super-resolution algorithms, such as the lack of texture-feature representation and of high-frequency details. Multi-scale detail enhancement can produce more texture information and high-frequency information. Therefore, super-resolution reconstruction of remote-sensing images based on adaptive multi-scale detail enhancement (AMDE-SR) is proposed in this paper. First, the information entropy of each remote-sensing image is calculated, and the image with the maximum entropy value is regarded as the reference image. Subsequently, spatio-temporal remote-sensing images are processed using phase normalization, which is to reduce the time phase difference of image data and enhance the complementarity of information. The multi-scale image information is then decomposed using the L ₀ gradient minimization model, and the non-redundant information is processed by difference calculation and expanding non-redundant layers and the redundant layer by the iterative back-projection (IBP) technique. The different-scale non-redundant information is adaptive-weighted and fused using cross-entropy. Finally, a nonlinear texture-detail-enhancement function is built to improve the scope of small details, and the peak signal-to-noise ratio (PSNR) is used as an iterative constraint. Ultimately, high-resolution remote-sensing images with abundant texture information are obtained by iterative optimization. Real results show an average gain in entropy of up to 0.42 dB for an up-scaling of 2 and a significant promotion gain in enhancement measure evaluation for an up-scaling of 2. The experimental results show that the performance of the AMED-SR method is better than existing super-resolution reconstruction methods in terms of visual and accuracy improvements.

  20. Spatio-Temporal Super-Resolution Reconstruction of Remote-Sensing Images Based on Adaptive Multi-Scale Detail Enhancement

    Science.gov (United States)

    Zhu, Hong; Tang, Xinming; Xie, Junfeng; Song, Weidong; Mo, Fan; Gao, Xiaoming

    2018-01-01

    There are many problems in existing reconstruction-based super-resolution algorithms, such as the lack of texture-feature representation and of high-frequency details. Multi-scale detail enhancement can produce more texture information and high-frequency information. Therefore, super-resolution reconstruction of remote-sensing images based on adaptive multi-scale detail enhancement (AMDE-SR) is proposed in this paper. First, the information entropy of each remote-sensing image is calculated, and the image with the maximum entropy value is regarded as the reference image. Subsequently, spatio-temporal remote-sensing images are processed using phase normalization, which is to reduce the time phase difference of image data and enhance the complementarity of information. The multi-scale image information is then decomposed using the L0 gradient minimization model, and the non-redundant information is processed by difference calculation and expanding non-redundant layers and the redundant layer by the iterative back-projection (IBP) technique. The different-scale non-redundant information is adaptive-weighted and fused using cross-entropy. Finally, a nonlinear texture-detail-enhancement function is built to improve the scope of small details, and the peak signal-to-noise ratio (PSNR) is used as an iterative constraint. Ultimately, high-resolution remote-sensing images with abundant texture information are obtained by iterative optimization. Real results show an average gain in entropy of up to 0.42 dB for an up-scaling of 2 and a significant promotion gain in enhancement measure evaluation for an up-scaling of 2. The experimental results show that the performance of the AMED-SR method is better than existing super-resolution reconstruction methods in terms of visual and accuracy improvements. PMID:29414893

  1. Estimation of reactogenicity of preparations produced on the basis of photoinactivated live vaccines against brucellosis and tularaemia on the organismic level.2. Using the method of speckle-microscopy with high spatial resolution

    Science.gov (United States)

    Ulianova, O. V.; Uianov, S. S.; Li, Pengcheng; Luo, Qingming

    2011-04-01

    The method of speckle microscopy was adapted to estimate the reactogenicity of the prototypes of vaccine preparations against extremely dangerous infections. The theory is proposed to describe the mechanism of formation of the output signal from the super-high spatial resolution speckle microscope. The experimental studies show that bacterial suspensions, irradiated in different regimes of inactivation, do not exert negative influence on the blood microcirculations in laboratory animals.

  2. Estimation of reactogenicity of preparations produced on the basis of photoinactivated live vaccines against brucellosis and tularaemia on the organismic level. 2. Using the method of speckle-microscopy with high spatial resolution

    International Nuclear Information System (INIS)

    Ulianova, O V; Uianov, S S; Li Pengcheng; Luo Qingming

    2011-01-01

    The method of speckle microscopy was adapted to estimate the reactogenicity of the prototypes of vaccine preparations against extremely dangerous infections. The theory is proposed to describe the mechanism of formation of the output signal from the super-high spatial resolution speckle microscope. The experimental studies show that bacterial suspensions, irradiated in different regimes of inactivation, do not exert negative influence on the blood microcirculations in laboratory animals. (optical technologies in biophysics and medicine)

  3. Novel Super-Resolution Approach to Time-Resolved Volumetric 4-Dimensional Magnetic Resonance Imaging With High Spatiotemporal Resolution for Multi-Breathing Cycle Motion Assessment

    Energy Technology Data Exchange (ETDEWEB)

    Li, Guang, E-mail: lig2@mskcc.org [Department of Medical Physics, Memorial Sloan Kettering Cancer Center, New York, New York (United States); Wei, Jie [Department of Computer Science, City College of New York, New York, New York (United States); Kadbi, Mo [Philips Healthcare, MR Therapy Cleveland, Ohio (United States); Moody, Jason; Sun, August; Zhang, Shirong; Markova, Svetlana; Zakian, Kristen; Hunt, Margie; Deasy, Joseph O. [Department of Medical Physics, Memorial Sloan Kettering Cancer Center, New York, New York (United States)

    2017-06-01

    Purpose: To develop and evaluate a super-resolution approach to reconstruct time-resolved 4-dimensional magnetic resonance imaging (TR-4DMRI) with a high spatiotemporal resolution for multi-breathing cycle motion assessment. Methods and Materials: A super-resolution approach was developed to combine fast 3-dimensional (3D) cine MRI with low resolution during free breathing (FB) and high-resolution 3D static MRI during breath hold (BH) using deformable image registration. A T1-weighted, turbo field echo sequence, coronal 3D cine acquisition, partial Fourier approximation, and SENSitivity Encoding parallel acceleration were used. The same MRI pulse sequence, field of view, and acceleration techniques were applied in both FB and BH acquisitions; the intensity-based Demons deformable image registration method was used. Under an institutional review board–approved protocol, 7 volunteers were studied with 3D cine FB scan (voxel size: 5 × 5 × 5 mm{sup 3}) at 2 Hz for 40 seconds and a 3D static BH scan (2 × 2 × 2 mm{sup 3}). To examine the image fidelity of 3D cine and super-resolution TR-4DMRI, a mobile gel phantom with multi-internal targets was scanned at 3 speeds and compared with the 3D static image. Image similarity among 3D cine, 4DMRI, and 3D static was evaluated visually using difference image and quantitatively using voxel intensity correlation and Dice index (phantom only). Multi-breathing-cycle waveforms were extracted and compared in both phantom and volunteer images using the 3D cine as the references. Results: Mild imaging artifacts were found in the 3D cine and TR-4DMRI of the mobile gel phantom with a Dice index of >0.95. Among 7 volunteers, the super-resolution TR-4DMRI yielded high voxel-intensity correlation (0.92 ± 0.05) and low voxel-intensity difference (<0.05). The detected motion differences between TR-4DMRI and 3D cine were −0.2 ± 0.5 mm (phantom) and −0.2 ± 1.9 mm (diaphragms). Conclusion: Super-resolution TR-4

  4. Novel Super-Resolution Approach to Time-Resolved Volumetric 4-Dimensional Magnetic Resonance Imaging With High Spatiotemporal Resolution for Multi-Breathing Cycle Motion Assessment

    International Nuclear Information System (INIS)

    Li, Guang; Wei, Jie; Kadbi, Mo; Moody, Jason; Sun, August; Zhang, Shirong; Markova, Svetlana; Zakian, Kristen; Hunt, Margie; Deasy, Joseph O.

    2017-01-01

    Purpose: To develop and evaluate a super-resolution approach to reconstruct time-resolved 4-dimensional magnetic resonance imaging (TR-4DMRI) with a high spatiotemporal resolution for multi-breathing cycle motion assessment. Methods and Materials: A super-resolution approach was developed to combine fast 3-dimensional (3D) cine MRI with low resolution during free breathing (FB) and high-resolution 3D static MRI during breath hold (BH) using deformable image registration. A T1-weighted, turbo field echo sequence, coronal 3D cine acquisition, partial Fourier approximation, and SENSitivity Encoding parallel acceleration were used. The same MRI pulse sequence, field of view, and acceleration techniques were applied in both FB and BH acquisitions; the intensity-based Demons deformable image registration method was used. Under an institutional review board–approved protocol, 7 volunteers were studied with 3D cine FB scan (voxel size: 5 × 5 × 5 mm"3) at 2 Hz for 40 seconds and a 3D static BH scan (2 × 2 × 2 mm"3). To examine the image fidelity of 3D cine and super-resolution TR-4DMRI, a mobile gel phantom with multi-internal targets was scanned at 3 speeds and compared with the 3D static image. Image similarity among 3D cine, 4DMRI, and 3D static was evaluated visually using difference image and quantitatively using voxel intensity correlation and Dice index (phantom only). Multi-breathing-cycle waveforms were extracted and compared in both phantom and volunteer images using the 3D cine as the references. Results: Mild imaging artifacts were found in the 3D cine and TR-4DMRI of the mobile gel phantom with a Dice index of >0.95. Among 7 volunteers, the super-resolution TR-4DMRI yielded high voxel-intensity correlation (0.92 ± 0.05) and low voxel-intensity difference (<0.05). The detected motion differences between TR-4DMRI and 3D cine were −0.2 ± 0.5 mm (phantom) and −0.2 ± 1.9 mm (diaphragms). Conclusion: Super-resolution TR-4DMRI has been

  5. Reducible Dictionaries for Single Image Super-Resolution based on Patch Matching and Mean Shifting

    DEFF Research Database (Denmark)

    Rasti, Pejman; Nasrollahi, Kamal; Orlova, Olga

    2017-01-01

    is taken, and its counterpart from the HR dictionary is passed through an illumination enhancement process. By this technique, the noticeable change of illumination between neighbor patches in the super-resolved image is significantly reduced. The enhanced HR patch represents the HR patch of the super......-resolved image. Finally, to remove the blocking effect caused by merging the patches, an average of the obtained HR image and the interpolated image obtained using bicubic interpolation is calculated. The quantitative and qualitative analyses show the superiority of the proposed technique over the conventional...

  6. Investigating sub-spine actin dynamics in rat hippocampal neurons with super-resolution optical imaging.

    Directory of Open Access Journals (Sweden)

    Vedakumar Tatavarty

    Full Text Available Morphological changes in dendritic spines represent an important mechanism for synaptic plasticity which is postulated to underlie the vital cognitive phenomena of learning and memory. These morphological changes are driven by the dynamic actin cytoskeleton that is present in dendritic spines. The study of actin dynamics in these spines traditionally has been hindered by the small size of the spine. In this study, we utilize a photo-activation localization microscopy (PALM-based single-molecule tracking technique to analyze F-actin movements with approximately 30-nm resolution in cultured hippocampal neurons. We were able to observe the kinematic (physical motion of actin filaments, i.e., retrograde flow and kinetic (F-actin turn-over dynamics of F-actin at the single-filament level in dendritic spines. We found that F-actin in dendritic spines exhibits highly heterogeneous kinematic dynamics at the individual filament level, with simultaneous actin flows in both retrograde and anterograde directions. At the ensemble level, movements of filaments integrate into a net retrograde flow of approximately 138 nm/min. These results suggest a weakly polarized F-actin network that consists of mostly short filaments in dendritic spines.

  7. Investigating sub-spine actin dynamics in rat hippocampal neurons with super-resolution optical imaging.

    Science.gov (United States)

    Tatavarty, Vedakumar; Kim, Eun-Ji; Rodionov, Vladimir; Yu, Ji

    2009-11-09

    Morphological changes in dendritic spines represent an important mechanism for synaptic plasticity which is postulated to underlie the vital cognitive phenomena of learning and memory. These morphological changes are driven by the dynamic actin cytoskeleton that is present in dendritic spines. The study of actin dynamics in these spines traditionally has been hindered by the small size of the spine. In this study, we utilize a photo-activation localization microscopy (PALM)-based single-molecule tracking technique to analyze F-actin movements with approximately 30-nm resolution in cultured hippocampal neurons. We were able to observe the kinematic (physical motion of actin filaments, i.e., retrograde flow) and kinetic (F-actin turn-over) dynamics of F-actin at the single-filament level in dendritic spines. We found that F-actin in dendritic spines exhibits highly heterogeneous kinematic dynamics at the individual filament level, with simultaneous actin flows in both retrograde and anterograde directions. At the ensemble level, movements of filaments integrate into a net retrograde flow of approximately 138 nm/min. These results suggest a weakly polarized F-actin network that consists of mostly short filaments in dendritic spines.

  8. Super-resolution convolutional neural network for the improvement of the image quality of magnified images in chest radiographs

    Science.gov (United States)

    Umehara, Kensuke; Ota, Junko; Ishimaru, Naoki; Ohno, Shunsuke; Okamoto, Kentaro; Suzuki, Takanori; Shirai, Naoki; Ishida, Takayuki

    2017-02-01

    Single image super-resolution (SR) method can generate a high-resolution (HR) image from a low-resolution (LR) image by enhancing image resolution. In medical imaging, HR images are expected to have a potential to provide a more accurate diagnosis with the practical application of HR displays. In recent years, the super-resolution convolutional neural network (SRCNN), which is one of the state-of-the-art deep learning based SR methods, has proposed in computer vision. In this study, we applied and evaluated the SRCNN scheme to improve the image quality of magnified images in chest radiographs. For evaluation, a total of 247 chest X-rays were sampled from the JSRT database. The 247 chest X-rays were divided into 93 training cases with non-nodules and 152 test cases with lung nodules. The SRCNN was trained using the training dataset. With the trained SRCNN, the HR image was reconstructed from the LR one. We compared the image quality of the SRCNN and conventional image interpolation methods, nearest neighbor, bilinear and bicubic interpolations. For quantitative evaluation, we measured two image quality metrics, peak signal-to-noise ratio (PSNR) and structural similarity (SSIM). In the SRCNN scheme, PSNR and SSIM were significantly higher than those of three interpolation methods (pmethods without any obvious artifacts. These preliminary results indicate that the SRCNN scheme significantly outperforms conventional interpolation algorithms for enhancing image resolution and that the use of the SRCNN can yield substantial improvement of the image quality of magnified images in chest radiographs.

  9. Super-resolution reconstruction of 4D-CT lung data via patch-based low-rank matrix reconstruction

    Science.gov (United States)

    Fang, Shiting; Wang, Huafeng; Liu, Yueliang; Zhang, Minghui; Yang, Wei; Feng, Qianjin; Chen, Wufan; Zhang, Yu

    2017-10-01

    Lung 4D computed tomography (4D-CT), which is a time-resolved CT data acquisition, performs an important role in explicitly including respiratory motion in treatment planning and delivery. However, the radiation dose is usually reduced at the expense of inter-slice spatial resolution to minimize radiation-related health risk. Therefore, resolution enhancement along the superior-inferior direction is necessary. In this paper, a super-resolution (SR) reconstruction method based on a patch low-rank matrix reconstruction is proposed to improve the resolution of lung 4D-CT images. Specifically, a low-rank matrix related to every patch is constructed by using a patch searching strategy. Thereafter, the singular value shrinkage is employed to recover the high-resolution patch under the constraints of the image degradation model. The output high-resolution patches are finally assembled to output the entire image. This method is extensively evaluated using two public data sets. Quantitative analysis shows that the proposed algorithm decreases the root mean square error by 9.7%-33.4% and the edge width by 11.4%-24.3%, relative to linear interpolation, back projection (BP) and Zhang et al’s algorithm. A new algorithm has been developed to improve the resolution of 4D-CT. In all experiments, the proposed method outperforms various interpolation methods, as well as BP and Zhang et al’s method, thus indicating the effectivity and competitiveness of the proposed algorithm.

  10. Stimulated Emission Pumping Enablling Sub-Diffraction-Limited Spatial Resolution in CARS Microscopy

    NARCIS (Netherlands)

    Cleff, C.; Gross, P.; Fallnich, C.; Offerhaus, Herman L.; Herek, Jennifer Lynn; Kruse, K.; Beeker, W.P; Beeker, W.P.; Lee, Christopher James; Boller, Klaus J.; Dobner, S.

    2012-01-01

    Suppression of CARS signal generation is demonstrated by equalization of the ground and Raman states via a control state in a theoretical investigation. Using donut-shaped control light fields for population transfer results in sub-diffraction-limited spatial resolution CARS microscopy.

  11. Gas-phase synthesis of magnesium nanoparticles : A high-resolution transmission electron microscopy study

    NARCIS (Netherlands)

    Kooi, B.J.; Palasantzas, G.; de Hosson, J.T.M.

    2006-01-01

    Magnesium nanoparticles with size above 10 nm, prepared by gas-phase syntheses, were investigated by high-resolution transmission electron microscopy. The dominant particle shape is a hexagonal prism terminated by Mg(0002) and Mg{1010} facets. Oxidation of Mg yields a MgO shell (similar to 3 nm

  12. Stacking it up: Exploring the limits of ultra-high resolution atomic force microscopy

    NARCIS (Netherlands)

    van der Heijden, N.J.

    2017-01-01

    Atomic force microscopy (AFM) is a technique wherein an atomically sharp needle raster scans across a surface, detecting forces between it and the sample. In state-of-the-art AFM experiments the measured forces are typically on the order of pico-Newtons, and the lateral resolution is on the order of

  13. Electron beam fabrication and characterization of high- resolution magnetic force microscopy tips

    NARCIS (Netherlands)

    Ruhrig, M.; Rührig, M.; Porthun, S.; Porthun, S.; Lodder, J.C.; Mc vitie, S.; Heyderman, L.J.; Johnston, A.B.; Chapman, J.N.

    1996-01-01

    The stray field, magnetic microstructure, and switching behavior of high‐resolution electron beam fabricated thin film tips for magnetic force microscopy (MFM) are investigated with different imaging modes in a transmission electron microscope (TEM). As the tiny smooth carbon needles covered with a

  14. Modelling high-resolution electron microscopy based on core-loss spectroscopy

    International Nuclear Information System (INIS)

    Allen, L.J.; Findlay, S.D.; Oxley, M.P.; Witte, C.; Zaluzec, N.J.

    2006-01-01

    There are a number of factors affecting the formation of images based on core-loss spectroscopy in high-resolution electron microscopy. We demonstrate unambiguously the need to use a full nonlocal description of the effective core-loss interaction for experimental results obtained from high angular resolution electron channelling electron spectroscopy. The implications of this model are investigated for atomic resolution scanning transmission electron microscopy. Simulations are used to demonstrate that core-loss spectroscopy images formed using fine probes proposed for future microscopes can result in images that do not correspond visually with the structure that has led to their formation. In this context, we also examine the effect of varying detector geometries. The importance of the contribution to core-loss spectroscopy images by dechannelled or diffusely scattered electrons is reiterated here

  15. Structured illumination microscopy and its new developments

    Directory of Open Access Journals (Sweden)

    Jianling Chen

    2016-05-01

    Full Text Available Optical microscopy allows us to observe the biological structures and processes within living cells. However, the spatial resolution of the optical microscopy is limited to about half of the wavelength by the light diffraction. Structured illumination microscopy (SIM, a type of new emerging super-resolution microscopy, doubles the spatial resolution by illuminating the specimen with a patterned light, and the sample and light source requirements of SIM are not as strict as the other super-resolution microscopy. In addition, SIM is easier to combine with the other imaging techniques to improve their imaging resolution, leading to the developments of diverse types of SIM. SIM has great potential to meet the various requirements of living cells imaging. Here, we review the recent developments of SIM and its combination with other imaging techniques.

  16. Super-resolution reconstruction in frequency, image, and wavelet domains to reduce through-plane partial voluming in MRI

    Energy Technology Data Exchange (ETDEWEB)

    Gholipour, Ali, E-mail: ali.gholipour@childrens.harvard.edu; Afacan, Onur; Scherrer, Benoit; Prabhu, Sanjay P.; Warfield, Simon K. [Department of Radiology, Boston Children’s Hospital, Boston, Massachusetts 02115 and Harvard Medical School, Boston, Massachusetts 02115 (United States); Aganj, Iman [Radiology Department, Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Boston, Massachusetts 02129 and Harvard Medical School, Boston, Massachusetts 02115 (United States); Sahin, Mustafa [Department of Neurology, Boston Children’s Hospital, Boston, Massachusetts 02115 and Harvard Medical School, Boston, Massachusetts 02115 (United States)

    2015-12-15

    Purpose: To compare and evaluate the use of super-resolution reconstruction (SRR), in frequency, image, and wavelet domains, to reduce through-plane partial voluming effects in magnetic resonance imaging. Methods: The reconstruction of an isotropic high-resolution image from multiple thick-slice scans has been investigated through techniques in frequency, image, and wavelet domains. Experiments were carried out with thick-slice T2-weighted fast spin echo sequence on the Academic College of Radiology MRI phantom, where the reconstructed images were compared to a reference high-resolution scan using peak signal-to-noise ratio (PSNR), structural similarity image metric (SSIM), mutual information (MI), and the mean absolute error (MAE) of image intensity profiles. The application of super-resolution reconstruction was then examined in retrospective processing of clinical neuroimages of ten pediatric patients with tuberous sclerosis complex (TSC) to reduce through-plane partial voluming for improved 3D delineation and visualization of thin radial bands of white matter abnormalities. Results: Quantitative evaluation results show improvements in all evaluation metrics through super-resolution reconstruction in the frequency, image, and wavelet domains, with the highest values obtained from SRR in the image domain. The metric values for image-domain SRR versus the original axial, coronal, and sagittal images were PSNR = 32.26 vs 32.22, 32.16, 30.65; SSIM = 0.931 vs 0.922, 0.924, 0.918; MI = 0.871 vs 0.842, 0.844, 0.831; and MAE = 5.38 vs 7.34, 7.06, 6.19. All similarity metrics showed high correlations with expert ranking of image resolution with MI showing the highest correlation at 0.943. Qualitative assessment of the neuroimages of ten TSC patients through in-plane and out-of-plane visualization of structures showed the extent of partial voluming effect in a real clinical scenario and its reduction using SRR. Blinded expert evaluation of image resolution in

  17. Super-resolution reconstruction in frequency, image, and wavelet domains to reduce through-plane partial voluming in MRI

    International Nuclear Information System (INIS)

    Gholipour, Ali; Afacan, Onur; Scherrer, Benoit; Prabhu, Sanjay P.; Warfield, Simon K.; Aganj, Iman; Sahin, Mustafa

    2015-01-01

    Purpose: To compare and evaluate the use of super-resolution reconstruction (SRR), in frequency, image, and wavelet domains, to reduce through-plane partial voluming effects in magnetic resonance imaging. Methods: The reconstruction of an isotropic high-resolution image from multiple thick-slice scans has been investigated through techniques in frequency, image, and wavelet domains. Experiments were carried out with thick-slice T2-weighted fast spin echo sequence on the Academic College of Radiology MRI phantom, where the reconstructed images were compared to a reference high-resolution scan using peak signal-to-noise ratio (PSNR), structural similarity image metric (SSIM), mutual information (MI), and the mean absolute error (MAE) of image intensity profiles. The application of super-resolution reconstruction was then examined in retrospective processing of clinical neuroimages of ten pediatric patients with tuberous sclerosis complex (TSC) to reduce through-plane partial voluming for improved 3D delineation and visualization of thin radial bands of white matter abnormalities. Results: Quantitative evaluation results show improvements in all evaluation metrics through super-resolution reconstruction in the frequency, image, and wavelet domains, with the highest values obtained from SRR in the image domain. The metric values for image-domain SRR versus the original axial, coronal, and sagittal images were PSNR = 32.26 vs 32.22, 32.16, 30.65; SSIM = 0.931 vs 0.922, 0.924, 0.918; MI = 0.871 vs 0.842, 0.844, 0.831; and MAE = 5.38 vs 7.34, 7.06, 6.19. All similarity metrics showed high correlations with expert ranking of image resolution with MI showing the highest correlation at 0.943. Qualitative assessment of the neuroimages of ten TSC patients through in-plane and out-of-plane visualization of structures showed the extent of partial voluming effect in a real clinical scenario and its reduction using SRR. Blinded expert evaluation of image resolution in

  18. Numerical study of the lateral resolution in electrostatic force microscopy for dielectric samples

    International Nuclear Information System (INIS)

    Riedel, C; AlegrIa, A; Colmenero, J; Schwartz, G A; Saenz, J J

    2011-01-01

    We present a study of the lateral resolution in electrostatic force microscopy for dielectric samples in both force and gradient modes. Whereas previous studies have reported expressions for metallic surfaces having potential heterogeneities (Kelvin probe force microscopy), in this work we take into account the presence of a dielectric medium. We introduce a definition of the lateral resolution based on the force due to a test particle being either a point charge or a polarizable particle on the dielectric surface. The behaviour has been studied over a wide range of typical experimental parameters: tip-sample distance (1-20) nm, sample thickness (0-5) μm and dielectric constant (1-20), using the numerical simulation of the equivalent charge method. For potential heterogeneities on metallic surfaces expressions are in agreement with the bibliography. The lateral resolution of samples having a dielectric constant of more than 10 tends to metallic behaviour. We found a characteristic thickness of 100 nm, above which the lateral resolution measured on the dielectric surface is close to that of an infinite medium. As previously reported, the lateral resolution is better in the gradient mode than in the force mode. Finally, we showed that for the same experimental conditions, the lateral resolution is better for a polarizable particle than for a charge, i.e. dielectric heterogeneities should always look 'sharper' (better resolved) than inhomogeneous charge distributions. This fact should be taken into account when interpreting images of heterogeneous samples.

  19. Visible light optical coherence microscopy imaging of the mouse cortex with femtoliter volume resolution

    Science.gov (United States)

    Merkle, Conrad W.; Chong, Shau Poh; Kho, Aaron M.; Zhu, Jun; Kholiqov, Oybek; Dubra, Alfredo; Srinivasan, Vivek J.

    2018-02-01

    Most flying-spot Optical Coherence Tomography (OCT) and Optical Coherence Microscopy (OCM) systems use a symmetric confocal geometry, where the detection path retraces the illumination path starting from and ending with the spatial mode of a single mode optical fiber. Here, we describe a visible light OCM instrument that breaks this symmetry to improve transverse resolution without sacrificing collection efficiency in scattering tissue. This was achieved by overfilling a 0.3 numerical aperture (NA) water immersion objective on the illumination path, while maintaining a conventional Gaussian mode detection path (1/e2 intensity diameter 0.82 Airy disks), enabling 1.1 μm full-width at half-maximum (FWHM) transverse resolution. At the same time, a 0.9 μm FWHM axial resolution in tissue, achieved by a broadband visible light source, enabled femtoliter volume resolution. We characterized this instrument according to paraxial coherent microscopy theory, and then used it to image the meningeal layers, intravascular red blood cell-free layer, and myelinated axons in the mouse neocortex in vivo through the thinned skull. Finally, by introducing a 0.8 NA water immersion objective, we improved the lateral resolution to 0.44 μm FWHM, which provided a volumetric resolution of 0.2 fL, revealing cell bodies in cortical layer I of the mouse brain with OCM for the first time.

  20. Numerical study of the lateral resolution in electrostatic force microscopy for dielectric samples

    Energy Technology Data Exchange (ETDEWEB)

    Riedel, C; AlegrIa, A; Colmenero, J [Departamento de Fisica de Materiales UPV/EHU, Facultad de Quimica, Apartado 1072, 20080 San Sebastian (Spain); Schwartz, G A [Centro de Fisica de Materiales CSIC-UPV/EHU, Paseo Manuel de Lardizabal 5, 20018 San Sebastian (Spain); Saenz, J J, E-mail: riedel@ies.univ-montp2.fr [Donostia International Physics Center, Paseo Manuel de Lardizabal 4, 20018 San Sebastian (Spain)

    2011-07-15

    We present a study of the lateral resolution in electrostatic force microscopy for dielectric samples in both force and gradient modes. Whereas previous studies have reported expressions for metallic surfaces having potential heterogeneities (Kelvin probe force microscopy), in this work we take into account the presence of a dielectric medium. We introduce a definition of the lateral resolution based on the force due to a test particle being either a point charge or a polarizable particle on the dielectric surface. The behaviour has been studied over a wide range of typical experimental parameters: tip-sample distance (1-20) nm, sample thickness (0-5) {mu}m and dielectric constant (1-20), using the numerical simulation of the equivalent charge method. For potential heterogeneities on metallic surfaces expressions are in agreement with the bibliography. The lateral resolution of samples having a dielectric constant of more than 10 tends to metallic behaviour. We found a characteristic thickness of 100 nm, above which the lateral resolution measured on the dielectric surface is close to that of an infinite medium. As previously reported, the lateral resolution is better in the gradient mode than in the force mode. Finally, we showed that for the same experimental conditions, the lateral resolution is better for a polarizable particle than for a charge, i.e. dielectric heterogeneities should always look 'sharper' (better resolved) than inhomogeneous charge distributions. This fact should be taken into account when interpreting images of heterogeneous samples.

  1. Adaptive patch-based POCS approach for super resolution reconstruction of 4D-CT lung data

    International Nuclear Information System (INIS)

    Wang, Tingting; Cao, Lei; Yang, Wei; Feng, Qianjin; Chen, Wufan; Zhang, Yu

    2015-01-01

    Image enhancement of lung four-dimensional computed tomography (4D-CT) data is highly important because image resolution remains a crucial point in lung cancer radiotherapy. In this paper, we proposed a method for lung 4D-CT super resolution (SR) by using an adaptive-patch-based projection onto convex sets (POCS) approach, which is in contrast with the global POCS SR algorithm, to recover fine details with lesser artifacts in images. The main contribution of this patch-based approach is that the interfering local structure from other phases can be rejected by employing a similar patch adaptive selection strategy. The effectiveness of our approach is demonstrated through experiments on simulated images and real lung 4D-CT datasets. A comparison with previously published SR reconstruction methods highlights the favorable characteristics of the proposed method. (paper)

  2. Wide-field two-dimensional multifocal optical-resolution photoacoustic computed microscopy

    Science.gov (United States)

    Xia, Jun; Li, Guo; Wang, Lidai; Nasiriavanaki, Mohammadreza; Maslov, Konstantin; Engelbach, John A.; Garbow, Joel R.; Wang, Lihong V.

    2014-01-01

    Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technique that directly images optical absorption in tissue at high spatial resolution. To date, the majority of OR-PAM systems are based on single focused optical excitation and ultrasonic detection, limiting the wide-field imaging speed. While one-dimensional multifocal OR-PAM (1D-MFOR-PAM) has been developed, the potential of microlens and transducer arrays has not been fully realized. Here, we present the development of two-dimensional multifocal optical-resolution photoacoustic computed microscopy (2D-MFOR-PACM), using a 2D microlens array and a full-ring ultrasonic transducer array. The 10 × 10 mm2 microlens array generates 1800 optical foci within the focal plane of the 512-element transducer array, and raster scanning the microlens array yields optical-resolution photoacoustic images. The system has improved the in-plane resolution of a full-ring transducer array from ≥100 µm to 29 µm and achieved an imaging time of 36 seconds over a 10 × 10 mm2 field of view. In comparison, the 1D-MFOR-PAM would take more than 4 minutes to image over the same field of view. The imaging capability of the system was demonstrated on phantoms and animals both ex vivo and in vivo. PMID:24322226

  3. SuperSegger

    DEFF Research Database (Denmark)

    Stylianidou, Stella; Brennan, Connor; Nissen, Silas B

    2016-01-01

    -colonies with many cells, facilitating the analysis of cell-cycle dynamics in bacteria as well as cell-contact mediated phenomena. This package has a range of built-in capabilities for characterizing bacterial cells, including the identification of cell division events, mother, daughter, and neighboring cells......Many quantitative cell biology questions require fast yet reliable automated image segmentation to identify and link cells from frame-to-frame, and characterize the cell morphology and fluorescence. We present SuperSegger, an automated MATLAB-based image processing package well......-suited to quantitative analysis of high-throughput live-cell fluorescence microscopy of bacterial cells. SuperSegger incorporates machine-learning algorithms to optimize cellular boundaries and automated error resolution to reliably link cells from frame-to-frame. Unlike existing packages, it can reliably segment micro...

  4. Super-resolution for everybody: An image processing workflow to obtain high-resolution images with a standard confocal microscope.

    Science.gov (United States)

    Lam, France; Cladière, Damien; Guillaume, Cyndélia; Wassmann, Katja; Bolte, Susanne

    2017-02-15

    In the presented work we aimed at improving confocal imaging to obtain highest possible resolution in thick biological samples, such as the mouse oocyte. We therefore developed an image processing workflow that allows improving the lateral and axial resolution of a standard confocal microscope. Our workflow comprises refractive index matching, the optimization of microscope hardware parameters and image restoration by deconvolution. We compare two different deconvolution algorithms, evaluate the necessity of denoising and establish the optimal image restoration procedure. We validate our workflow by imaging sub resolution fluorescent beads and measuring the maximum lateral and axial resolution of the confocal system. Subsequently, we apply the parameters to the imaging and data restoration of fluorescently labelled meiotic spindles of mouse oocytes. We measure a resolution increase of approximately 2-fold in the lateral and 3-fold in the axial direction throughout a depth of 60μm. This demonstrates that with our optimized workflow we reach a resolution that is comparable to 3D-SIM-imaging, but with better depth penetration for confocal images of beads and the biological sample. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. A Fast Algorithm for Image Super-Resolution from Blurred Observations

    Directory of Open Access Journals (Sweden)

    Ng Michael K

    2006-01-01

    Full Text Available We study the problem of reconstruction of a high-resolution image from several blurred low-resolution image frames. The image frames consist of blurred, decimated, and noisy versions of a high-resolution image. The high-resolution image is modeled as a Markov random field (MRF, and a maximum a posteriori (MAP estimation technique is used for the restoration. We show that with the periodic boundary condition, a high-resolution image can be restored efficiently by using fast Fourier transforms. We also apply the preconditioned conjugate gradient method to restore high-resolution images in the aperiodic boundary condition. Computer simulations are given to illustrate the effectiveness of the proposed approach.

  6. Fluorescent Nanodiamond: A Versatile Tool for Long-Term Cell Tracking, Super-Resolution Imaging, and Nanoscale Temperature Sensing.

    Science.gov (United States)

    Hsiao, Wesley Wei-Wen; Hui, Yuen Yung; Tsai, Pei-Chang; Chang, Huan-Cheng

    2016-03-15

    Fluorescent nanodiamond (FND) has recently played a central role in fueling new discoveries in interdisciplinary fields spanning biology, chemistry, physics, and materials sciences. The nanoparticle is unique in that it contains a high density ensemble of negatively charged nitrogen-vacancy (NV(-)) centers as built-in fluorophores. The center possesses a number of outstanding optical and magnetic properties. First, NV(-) has an absorption maximum at ∼550 nm, and when exposed to green-orange light, it emits bright fluorescence at ∼700 nm with a lifetime of longer than 10 ns. These spectroscopic properties are little affected by surface modification but are distinctly different from those of cell autofluorescence and thus enable background-free imaging of FNDs in tissue sections. Such characteristics together with its excellent biocompatibility render FND ideal for long-term cell tracking applications, particularly in stem cell research. Next, as an artificial atom in the solid state, the NV(-) center is perfectly photostable, without photobleaching and blinking. Therefore, the NV-containing FND is suitable as a contrast agent for super-resolution imaging by stimulated emission depletion (STED). An improvement of the spatial resolution by 20-fold is readily achievable by using a high-power STED laser to deplete the NV(-) fluorescence. Such improvement is crucial in revealing the detailed structures of biological complexes and assemblies, including cellular organelles and subcellular compartments. Further enhancement of the resolution for live cell imaging is possible by manipulating the charge states of the NV centers. As the "brightest" member of the nanocarbon family, FND holds great promise and potential for bioimaging with unprecedented resolution and precision. Lastly, the NV(-) center in diamond is an atom-like quantum system with a total electron spin of 1. The ground states of the spins show a crystal field splitting of 2.87 GHz, separating the ms = 0 and

  7. Advanced Microscopy of Microbial Cells

    DEFF Research Database (Denmark)

    Haagensen, Janus Anders Juul; Regenberg, Birgitte; Sternberg, Claus

    2011-01-01

    Growing awareness of heterogeneity in cells of microbial populations has emphasized the importance of advanced microscopy for visualization and understanding of the molecular mechanisms underlying cell-to-cell variation. In this review, we highlight some of the recent advances in confocal...... microscopy, super-resolution optical microscopy (STED, SIM, PALM) as well as atomic force microscopy and Raman spectroscopy. Using examples of bistability in microbial populations as well as biofilm development and differentiation in bacterial and yeast consortia, we demonstrate the importance of microscopy...

  8. Determining the resolution of scanning microwave impedance microscopy using atomic-precision buried donor structures

    Science.gov (United States)

    Scrymgeour, D. A.; Baca, A.; Fishgrab, K.; Simonson, R. J.; Marshall, M.; Bussmann, E.; Nakakura, C. Y.; Anderson, M.; Misra, S.

    2017-11-01

    To quantify the resolution limits of scanning microwave impedance microscopy (sMIM), we created scanning tunneling microscope (STM)-patterned donor nanostructures in silicon composed of 10 nm lines of highly conductive silicon buried under a protective top cap of silicon, and imaged them with sMIM. This dopant pattern is an ideal test of the resolution and sensitivity of the sMIM technique, as it is made with nm-resolution and offers minimal complications from topography convolution. It has been determined that typical sMIM tips can resolve lines down to ∼80 nm spacing, while resolution is independent of tip geometry as extreme tip wear does not change the resolving power, contrary to traditional scanning capacitance microscopy (SCM). Going forward, sMIM is an ideal technique for qualifying buried patterned devices, potentially allowing for quantitative post-fabrication characterization of donor structures, which may be an important tool for the study of atomic-scale transistors and state of the art quantum computation schemes.

  9. Atomic-resolution transmission electron microscopy of electron beam–sensitive crystalline materials

    Science.gov (United States)

    Zhang, Daliang; Zhu, Yihan; Liu, Lingmei; Ying, Xiangrong; Hsiung, Chia-En; Sougrat, Rachid; Li, Kun; Han, Yu

    2018-02-01

    High-resolution imaging of electron beam–sensitive materials is one of the most difficult applications of transmission electron microscopy (TEM). The challenges are manifold, including the acquisition of images with extremely low beam doses, the time-constrained search for crystal zone axes, the precise image alignment, and the accurate determination of the defocus value. We develop a suite of methods to fulfill these requirements and acquire atomic-resolution TEM images of several metal organic frameworks that are generally recognized as highly sensitive to electron beams. The high image resolution allows us to identify individual metal atomic columns, various types of surface termination, and benzene rings in the organic linkers. We also apply our methods to other electron beam–sensitive materials, including the organic-inorganic hybrid perovskite CH3NH3PbBr3.

  10. Atomic-resolution transmission electron microscopy of electron beam–sensitive crystalline materials

    KAUST Repository

    Zhang, Daliang

    2018-01-18

    High-resolution imaging of electron beam-sensitive materials is one of the most difficult applications of transmission electron microscopy (TEM). The challenges are manifold, including the acquisition of images with extremely low beam doses, the time-constrained search for crystal zone axes, the precise image alignment, and the accurate determination of the defocus value. We develop a suite of methods to fulfill these requirements and acquire atomic-resolution TEM images of several metal organic frameworks that are generally recognized as highly sensitive to electron beams. The high image resolution allows us to identify individual metal atomic columns, various types of surface termination, and benzene rings in the organic linkers. We also apply our methods to other electron beam–sensitive materials, including the organic-inorganic hybrid perovskite CH3NH3PbBr3.

  11. Rapid increase of near atomic resolution virus capsid structures determined by cryo-electron microscopy.

    Science.gov (United States)

    Ho, Phuong T; Reddy, Vijay S

    2018-01-01

    The recent technological advances in electron microscopes, detectors, as well as image processing and reconstruction software have brought single particle cryo-electron microscopy (cryo-EM) into prominence for determining structures of bio-molecules at near atomic resolution. This has been particularly true for virus capsids, ribosomes, and other large assemblies, which have been the ideal specimens for structural studies by cryo-EM approaches. An analysis of time series metadata of virus structures on the methods of structure determination, resolution of the structures, and size of the virus particles revealed a rapid increase in the virus structures determined by cryo-EM at near atomic resolution since 2010. In addition, the data highlight the median resolution (∼3.0 Å) and size (∼310.0 Å in diameter) of the virus particles determined by X-ray crystallography while no such limits exist for cryo-EM structures, which have a median diameter of 508 Å. Notably, cryo-EM virus structures in the last four years have a median resolution of 3.9 Å. Taken together with minimal sample requirements, not needing diffraction quality crystals, and being able to achieve similar resolutions of the crystal structures makes cryo-EM the method of choice for current and future virus capsid structure determinations. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Super-resolution post-processing for satellites with yaw-steering capability

    CSIR Research Space (South Africa)

    Van den Dool, R

    2012-10-01

    Full Text Available We describe a method for improving Earth observation satellite image resolution, for specific areas of interest where the sensor design resolution is insufficient. Our method may be used for satellites with yaw-steering capability, such as Nigeria...

  13. Application of carbon nanotubes to topographical resolution enhancement of tapered fiber scanning near field optical microscopy probes

    Science.gov (United States)

    Huntington, S. T.; Jarvis, S. P.

    2003-05-01

    Scanning near field optical microscopy (SNOM) probes are typically tapered optical fibers with metallic coatings. The tip diameters are generally in excess of 300 nm and thus provide poor topographical resolution. Here we report on the attachment multiwalled carbon nanotubes to the probes in order to substantially enhance the topographical resolution, without adversely affecting the optical resolution.

  14. Super-resolution nanofabrication with metal-ion doped hybrid material through an optical dual-beam approach

    International Nuclear Information System (INIS)

    Cao, Yaoyu; Li, Xiangping; Gu, Min

    2014-01-01

    We apply an optical dual-beam approach to a metal-ion doped hybrid material to achieve nanofeatures beyond the optical diffraction limit. By spatially inhibiting the photoreduction and the photopolymerization, we realize a nano-line, consisting of polymer matrix and in-situ generated gold nanoparticles, with a lateral size of sub 100 nm, corresponding to a factor of 7 improvement compared to the diffraction limit. With the existence of gold nanoparticles, a plasmon enhanced super-resolution fabrication mechanism in the hybrid material is observed, which benefits in a further reduction in size of the fabricated feature. The demonstrated nanofeature in hybrid materials paves the way for realizing functional nanostructures

  15. A numerical study of super-resolution through fast 3D wideband algorithm for scattering in highly-heterogeneous media

    KAUST Repository

    Létourneau, Pierre-David

    2016-09-19

    We present a wideband fast algorithm capable of accurately computing the full numerical solution of the problem of acoustic scattering of waves by multiple finite-sized bodies such as spherical scatterers in three dimensions. By full solution, we mean that no assumption (e.g. Rayleigh scattering, geometrical optics, weak scattering, Born single scattering, etc.) is necessary regarding the properties of the scatterers, their distribution or the background medium. The algorithm is also fast in the sense that it scales linearly with the number of unknowns. We use this algorithm to study the phenomenon of super-resolution in time-reversal refocusing in highly-scattering media recently observed experimentally (Lemoult et al., 2011), and provide numerical arguments towards the fact that such a phenomenon can be explained through a homogenization theory.

  16. Flat super-oscillatory lens for heat-assisted magnetic recording with sub-50 nm resolution.

    Science.gov (United States)

    Yuan, Guanghui; Rogers, Edward T F; Roy, Tapashree; Shen, Zexiang; Zheludev, Nikolay I

    2014-03-24

    Heat-assisted magnetic recording (HAMR) is a future roadmap technology to overcome the superparamagnetic limit in high density magnetic recording. Existing HAMR schemes depend on a simultaneous magnetic stimulation and light-induced local heating of the information carrier. To achieve high-density recorded data, near-field plasmonic transducers have been proposed as light concentrators. Here we suggest and investigate in detail an alternative approach exploiting a far-field focusing device that can focus light into sub-50 nm hot-spots in the magnetic recording layer using a laser source operating at 473 nm. It is based on a recently introduced super-oscillatory flat lens improved with the use of solid immersion, giving an effective numerical aperture as high as 4.17. The proposed solution is robust and easy to integrate with the magnetic recording head thus offering a competitive advantage over plasmonic technology.

  17. Effective deep learning training for single-image super-resolution in endomicroscopy exploiting video-registration-based reconstruction.

    Science.gov (United States)

    Ravì, Daniele; Szczotka, Agnieszka Barbara; Shakir, Dzhoshkun Ismail; Pereira, Stephen P; Vercauteren, Tom

    2018-06-01

    Probe-based confocal laser endomicroscopy (pCLE) is a recent imaging modality that allows performing in vivo optical biopsies. The design of pCLE hardware, and its reliance on an optical fibre bundle, fundamentally limits the image quality with a few tens of thousands fibres, each acting as the equivalent of a single-pixel detector, assembled into a single fibre bundle. Video registration techniques can be used to estimate high-resolution (HR) images by exploiting the temporal information contained in a sequence of low-resolution (LR) images. However, the alignment of LR frames, required for the fusion, is computationally demanding and prone to artefacts. In this work, we propose a novel synthetic data generation approach to train exemplar-based Deep Neural Networks (DNNs). HR pCLE images with enhanced quality are recovered by the models trained on pairs of estimated HR images (generated by the video registration algorithm) and realistic synthetic LR images. Performance of three different state-of-the-art DNNs techniques were analysed on a Smart Atlas database of 8806 images from 238 pCLE video sequences. The results were validated through an extensive image quality assessment that takes into account different quality scores, including a Mean Opinion Score (MOS). Results indicate that the proposed solution produces an effective improvement in the quality of the obtained reconstructed image. The proposed training strategy and associated DNNs allows us to perform convincing super-resolution of pCLE images.

  18. Improving axial resolution in confocal microscopy with new high refractive index mounting media.

    Science.gov (United States)

    Fouquet, Coralie; Gilles, Jean-François; Heck, Nicolas; Dos Santos, Marc; Schwartzmann, Richard; Cannaya, Vidjeacoumary; Morel, Marie-Pierre; Davidson, Robert Stephen; Trembleau, Alain; Bolte, Susanne

    2015-01-01

    Resolution, high signal intensity and elevated signal to noise ratio (SNR) are key issues for biologists who aim at studying the localisation of biological structures at the cellular and subcellular levels using confocal microscopy. The resolution required to separate sub-cellular biological structures is often near to the resolving power of the microscope. When optimally used, confocal microscopes may reach resolutions of 180 nm laterally and 500 nm axially, however, axial resolution in depth is often impaired by spherical aberration that may occur due to refractive index mismatches. Spherical aberration results in broadening of the point-spread function (PSF), a decrease in peak signal intensity when imaging in depth and a focal shift that leads to the distortion of the image along the z-axis and thus in a scaling error. In this study, we use the novel mounting medium CFM3 (Citifluor Ltd., UK) with a refractive index of 1.518 to minimize the effects of spherical aberration. This mounting medium is compatible with most common fluorochromes and fluorescent proteins. We compare its performance with established mounting media, harbouring refractive indices below 1.500, by estimating lateral and axial resolution with sub-resolution fluorescent beads. We show furthermore that the use of the high refractive index media renders the tissue transparent and improves considerably the axial resolution and imaging depth in immuno-labelled or fluorescent protein labelled fixed mouse brain tissue. We thus propose to use those novel high refractive index mounting media, whenever optimal axial resolution is required.

  19. Improving axial resolution in confocal microscopy with new high refractive index mounting media.

    Directory of Open Access Journals (Sweden)

    Coralie Fouquet

    Full Text Available Resolution, high signal intensity and elevated signal to noise ratio (SNR are key issues for biologists who aim at studying the localisation of biological structures at the cellular and subcellular levels using confocal microscopy. The resolution required to separate sub-cellular biological structures is often near to the resolving power of the microscope. When optimally used, confocal microscopes may reach resolutions of 180 nm laterally and 500 nm axially, however, axial resolution in depth is often impaired by spherical aberration that may occur due to refractive index mismatches. Spherical aberration results in broadening of the point-spread function (PSF, a decrease in peak signal intensity when imaging in depth and a focal shift that leads to the distortion of the image along the z-axis and thus in a scaling error. In this study, we use the novel mounting medium CFM3 (Citifluor Ltd., UK with a refractive index of 1.518 to minimize the effects of spherical aberration. This mounting medium is compatible with most common fluorochromes and fluorescent proteins. We compare its performance with established mounting media, harbouring refractive indices below 1.500, by estimating lateral and axial resolution with sub-resolution fluorescent beads. We show furthermore that the use of the high refractive index media renders the tissue transparent and improves considerably the axial resolution and imaging depth in immuno-labelled or fluorescent protein labelled fixed mouse brain tissue. We thus propose to use those novel high refractive index mounting media, whenever optimal axial resolution is required.

  20. High-spatial-resolution sub-surface imaging using a laser-based acoustic microscopy technique.

    Science.gov (United States)

    Balogun, Oluwaseyi; Cole, Garrett D; Huber, Robert; Chinn, Diane; Murray, Todd W; Spicer, James B

    2011-01-01

    Scanning acoustic microscopy techniques operating at frequencies in the gigahertz range are suitable for the elastic characterization and interior imaging of solid media with micrometer-scale spatial resolution. Acoustic wave propagation at these frequencies is strongly limited by energy losses, particularly from attenuation in the coupling media used to transmit ultrasound to a specimen, leading to a decrease in the depth in a specimen that can be interrogated. In this work, a laser-based acoustic microscopy technique is presented that uses a pulsed laser source for the generation of broadband acoustic waves and an optical interferometer for detection. The use of a 900-ps microchip pulsed laser facilitates the generation of acoustic waves with frequencies extending up to 1 GHz which allows for the resolution of micrometer-scale features in a specimen. Furthermore, the combination of optical generation and detection approaches eliminates the use of an ultrasonic coupling medium, and allows for elastic characterization and interior imaging at penetration depths on the order of several hundred micrometers. Experimental results illustrating the use of the laser-based acoustic microscopy technique for imaging micrometer-scale subsurface geometrical features in a 70-μm-thick single-crystal silicon wafer with a (100) orientation are presented.

  1. High-resolution photoluminescence electro-modulation microscopy by scanning lock-in

    Science.gov (United States)

    Koopman, W.; Muccini, M.; Toffanin, S.

    2018-04-01

    Morphological inhomogeneities and structural defects in organic semiconductors crucially determine the charge accumulation and lateral transport in organic thin-film transistors. Photoluminescence Electro-Modulation (PLEM) microscopy is a laser-scanning microscopy technique that relies on the modulation of the thin-film fluorescence in the presence of charge-carriers to image the spatial distribution of charges within the active organic semiconductor. Here, we present a lock-in scheme based on a scanning beam approach for increasing the PLEM microscopy resolution and contrast. The charge density in the device is modulated by a sinusoidal electrical signal, phase-locked to the scanning beam of the excitation laser. The lock-in detection scheme is achieved by acquiring a series of images with different phases between the beam scan and the electrical modulation. Application of high resolution PLEM to an organic transistor in accumulation mode demonstrates its potential to image local variations in the charge accumulation. A diffraction-limited precision of sub-300 nm and a signal to noise ratio of 21.4 dB could be achieved.

  2. Projections onto Convex Sets Super-Resolution Reconstruction Based on Point Spread Function Estimation of Low-Resolution Remote Sensing Images

    Directory of Open Access Journals (Sweden)

    Chong Fan

    2017-02-01

    Full Text Available To solve the problem on inaccuracy when estimating the point spread function (PSF of the ideal original image in traditional projection onto convex set (POCS super-resolution (SR reconstruction, this paper presents an improved POCS SR algorithm based on PSF estimation of low-resolution (LR remote sensing images. The proposed algorithm can improve the spatial resolution of the image and benefit agricultural crop visual interpolation. The PSF of the highresolution (HR image is unknown in reality. Therefore, analysis of the relationship between the PSF of the HR image and the PSF of the LR image is important to estimate the PSF of the HR image by using multiple LR images. In this study, the linear relationship between the PSFs of the HR and LR images can be proven. In addition, the novel slant knife-edge method is employed, which can improve the accuracy of the PSF estimation of LR images. Finally, the proposed method is applied to reconstruct airborne digital sensor 40 (ADS40 three-line array images and the overlapped areas of two adjacent GF-2 images by embedding the estimated PSF of the HR image to the original POCS SR algorithm. Experimental results show that the proposed method yields higher quality of reconstructed images than that produced by the blind SR method and the bicubic interpolation method.

  3. Super resolution reconstruction of infrared images based on classified dictionary learning

    Science.gov (United States)

    Liu, Fei; Han, Pingli; Wang, Yi; Li, Xuan; Bai, Lu; Shao, Xiaopeng

    2018-05-01

    Infrared images always suffer from low-resolution problems resulting from limitations of imaging devices. An economical approach to combat this problem involves reconstructing high-resolution images by reasonable methods without updating devices. Inspired by compressed sensing theory, this study presents and demonstrates a Classified Dictionary Learning method to reconstruct high-resolution infrared images. It classifies features of the samples into several reasonable clusters and trained a dictionary pair for each cluster. The optimal pair of dictionaries is chosen for each image reconstruction and therefore, more satisfactory results is achieved without the increase in computational complexity and time cost. Experiments and results demonstrated that it is a viable method for infrared images reconstruction since it improves image resolution and recovers detailed information of targets.

  4. Sparsity-Based Pixel Super Resolution for Lens-Free Digital In-line Holography.

    Science.gov (United States)

    Song, Jun; Leon Swisher, Christine; Im, Hyungsoon; Jeong, Sangmoo; Pathania, Divya; Iwamoto, Yoshiko; Pivovarov, Misha; Weissleder, Ralph; Lee, Hakho

    2016-04-21

    Lens-free digital in-line holography (LDIH) is a promising technology for portable, wide field-of-view imaging. Its resolution, however, is limited by the inherent pixel size of an imaging device. Here we present a new computational approach to achieve sub-pixel resolution for LDIH. The developed method is a sparsity-based reconstruction with the capability to handle the non-linear nature of LDIH. We systematically characterized the algorithm through simulation and LDIH imaging studies. The method achieved the spatial resolution down to one-third of the pixel size, while requiring only single-frame imaging without any hardware modifications. This new approach can be used as a general framework to enhance the resolution in nonlinear holographic systems.

  5. Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging

    Czech Academy of Sciences Publication Activity Database

    Lukeš, T.; Glatzová, Daniela; Kvíčalová, Zuzana; Levet, F.; Benda, Aleš; Letschert, S.; Sauer, M.; Brdička, Tomáš; Lasser, T.; Cebecauer, Marek

    2017-01-01

    Roč. 8, č. 1 (2017), č. článku 1731. ISSN 2041-1723 R&D Projects: GA ČR GA15-06989S Institutional support: RVO:61388955 ; RVO:68378050 Keywords : quantifying protein densities * membranes * single-molecule localization microscopy Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 12.124, year: 2016

  6. A resolution study for electrostatic force microscopy on bimetallic samples using the boundary element method

    International Nuclear Information System (INIS)

    Shen Yongxing; Lee, Minhwan; Lee, Wonyoung; Barnett, David M; Pinsky, Peter M; Prinz, Friedrich B

    2008-01-01

    Electrostatic force microscopy (EFM) is a special design of non-contact atomic force microscopy used for detecting electrostatic interactions between the probe tip and the sample. Its resolution is limited by the finite probe size and the long-range characteristics of electrostatic forces. Therefore, quantitative analysis is crucial to understanding the relationship between the actual local surface potential distribution and the quantities obtained from EFM measurements. To study EFM measurements on bimetallic samples with surface potential inhomogeneities as a special case, we have simulated such measurements using the boundary element method and calculated the force component and force gradient component that would be measured by amplitude modulation (AM) EFM and frequency modulation (FM) EFM, respectively. Such analyses have been performed for inhomogeneities of various shapes and sizes, for different tip-sample separations and tip geometries, for different applied voltages, and for different media (e.g., vacuum or water) in which the experiment is performed. For a sample with a surface potential discontinuity, the FM-EFM resolution expression agrees with the literature; however, the simulation for AM-EFM suggests the existence of an optimal tip radius of curvature in terms of resolution. On the other hand, for samples with strip- and disk-shaped surface potential inhomogeneities, we have obtained quantitative expressions for the detectability size requirements as a function of experimental conditions for both AM- and FM-EFMs, which suggest that a larger tip radius of curvature is moderately favored for detecting the presence of such inhomogeneities

  7. Fast time-resolved electrostatic force microscopy: Achieving sub-cycle time resolution

    Energy Technology Data Exchange (ETDEWEB)

    Karatay, Durmus U.; Harrison, Jeffrey S.; Glaz, Micah S.; Giridharagopal, Rajiv; Ginger, David S., E-mail: ginger@chem.washington.edu [Department of Chemistry, University of Washington, Seattle, Washington 98195 (United States)

    2016-05-15

    The ability to measure microsecond- and nanosecond-scale local dynamics below the diffraction limit with widely available atomic force microscopy hardware would enable new scientific studies in fields ranging from biology to semiconductor physics. However, commercially available scanning-probe instruments typically offer the ability to measure dynamics only on time scales of milliseconds to seconds. Here, we describe in detail the implementation of fast time-resolved electrostatic force microscopy using an oscillating cantilever as a means to measure fast local dynamics following a perturbation to a sample. We show how the phase of the oscillating cantilever relative to the perturbation event is critical to achieving reliable sub-cycle time resolution. We explore how noise affects the achievable time resolution and present empirical guidelines for reducing noise and optimizing experimental parameters. Specifically, we show that reducing the noise on the cantilever by using photothermal excitation instead of piezoacoustic excitation further improves time resolution. We demonstrate the discrimination of signal rise times with time constants as fast as 10 ns, and simultaneous data acquisition and analysis for dramatically improved image acquisition times.

  8. Tip radius preservation for high resolution imaging in amplitude modulation atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Ramos, Jorge R., E-mail: jorge.rr@cea.cu [Instituto de Ciencia de Materiales de Madrid, Sor Juana Inés de la Cruz 3, Canto Blanco, 28049 Madrid, España (Spain)

    2014-07-28

    The acquisition of high resolution images in atomic force microscopy (AFM) is correlated to the cantilever's tip shape, size, and imaging conditions. In this work, relative tip wear is quantified based on the evolution of a direct experimental observable in amplitude modulation atomic force microscopy, i.e., the critical amplitude. We further show that the scanning parameters required to guarantee a maximum compressive stress that is lower than the yield/fracture stress of the tip can be estimated via experimental observables. In both counts, the optimized parameters to acquire AFM images while preserving the tip are discussed. The results are validated experimentally by employing IgG antibodies as a model system.

  9. A high resolution electron microscopy investigation of curvature in multilayer graphite sheets

    International Nuclear Information System (INIS)

    Wang Zhenxia; Hu Jun; Wang Wenmin; Yu Guoqing

    1998-01-01

    Here the authors report a carbon sample generated by ultrasonic wave high oriented pyrolytic graphite (HOPG) in ethanol, water or ethanol-water mixed solution. High resolution transmission electron microscopy (HRTEM) revealed many multilayer graphite sheets with a total curved angle that is multiples of θ 0 (= 30 degree C). Close examination of the micrographs showed that the curvature is accomplished by bending the lattice planes. A possible explanation for the curvature in multilayer graphite sheets is discussed based on the conformation of graphite symmetry axes and the formation of sp 3 -like line defects in the sp 2 graphitic network

  10. Fluorescent Rhodamines and Fluorogenic Carbopyronines for Super‐Resolution STED Microscopy in Living Cells

    Science.gov (United States)

    Mitronova, Gyuzel Yu.; Sidenstein, Sven C.; Klocke, Jessica L.; Kamin, Dirk; Meineke, Dirk N. H.; D'Este, Elisa; Kraemer, Philip‐Tobias; Danzl, Johann G.

    2016-01-01

    Abstract A range of bright and photostable rhodamines and carbopyronines with absorption maxima in the range of λ=500–630 nm were prepared, and enabled the specific labeling of cytoskeletal filaments using HaloTag technology followed by staining with 1 μm solutions of the dye–ligand conjugates. The synthesis, photophysical parameters, fluorogenic behavior, and structure–property relationships of the new dyes are discussed. Light microscopy with stimulated emission depletion (STED) provided one‐ and two‐color images of living cells with an optical resolution of 40–60 nm. PMID:26844929

  11. Gas-phase synthesis of magnesium nanoparticles: A high-resolution transmission electron microscopy study

    International Nuclear Information System (INIS)

    Kooi, B. J.; Palasantzas, G.; De Hosson, J. Th. M.

    2006-01-01

    Magnesium nanoparticles with size above 10 nm, prepared by gas-phase syntheses, were investigated by high-resolution transmission electron microscopy. The dominant particle shape is a hexagonal prism terminated by Mg(0002) and Mg(1010) facets. Oxidation of Mg yields a MgO shell (∼3 nm thick), which has an orientation relation with the Mg. Inhomogeneous facet oxidation influences their growth kinetics resulting in a relatively broad size and shape distribution. Faceted voids between Mg and MgO shells indicate a fast outward diffusion of Mg and vacancy rearrangement into voids. The faceting of polar (220) planes is assisted by electron irradiation

  12. Exploring semiconductor quantum dots and wires by high resolution electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Molina, S I [Departamento de Ciencia de los Materiales e Ing Metalurgica y Q. Inorganica, F. de Ciencias, Universidad de Cadiz, Campus Rio San Pedro. 11510 Puerto Real (Cadiz) (Spain); Galindo, P L [Departamento de Lenguajes y Sistemas Informaticos, CASEM, Universidad de Cadiz, Campus Rio San Pedro. 11510 Puerto Real (Cadiz) (Spain); Gonzalez, L; Ripalda, J M [Instituto de Microelectronica de Madrid (CNM, CSIC), Isaac Newton 8, 28760 Tres Cantos, Madrid (Spain); Varela, M; Pennycook, S J, E-mail: sergio.molina@uca.e [Materials Science and Technology Division, Oak Ridge National Laboratory, Oak Ridge TN 37831 (United States)

    2010-02-01

    We review in this communication our contribution to the structural characterisation of semiconductor quantum dots and wires by high resolution electron microscopy, both in phase-contrast and Z-contrast modes. We show how these techniques contribute to predict the preferential sites of nucleation of these nanostructures, and also determine the compositional distribution in 1D and 0D nanostructures. The results presented here were produced in the framework of the European Network of Excellence entitled {sup S}elf-Assembled semiconductor Nanostructures for new Devices in photonics and Electronics (SANDiE){sup .}

  13. Single image super resolution algorithm based on edge interpolation in NSCT domain

    Science.gov (United States)

    Zhang, Mengqun; Zhang, Wei; He, Xinyu

    2017-11-01

    In order to preserve the texture and edge information and to improve the space resolution of single frame, a superresolution algorithm based on Contourlet (NSCT) is proposed. The original low resolution image is transformed by NSCT, and the directional sub-band coefficients of the transform domain are obtained. According to the scale factor, the high frequency sub-band coefficients are amplified by the interpolation method based on the edge direction to the desired resolution. For high frequency sub-band coefficients with noise and weak targets, Bayesian shrinkage is used to calculate the threshold value. The coefficients below the threshold are determined by the correlation among the sub-bands of the same scale to determine whether it is noise and de-noising. The anisotropic diffusion filter is used to effectively enhance the weak target in the low contrast region of the target and background. Finally, the high-frequency sub-band is amplified by the bilinear interpolation method to the desired resolution, and then combined with the high-frequency subband coefficients after de-noising and small target enhancement, the NSCT inverse transform is used to obtain the desired resolution image. In order to verify the effectiveness of the proposed algorithm, the proposed algorithm and several common image reconstruction methods are used to test the synthetic image, motion blurred image and hyperspectral image, the experimental results show that compared with the traditional single resolution algorithm, the proposed algorithm can obtain smooth edges and good texture features, and the reconstructed image structure is well preserved and the noise is suppressed to some extent.

  14. Functional magnetic resonance microscopy at single-cell resolution in Aplysia californica

    Science.gov (United States)

    Radecki, Guillaume; Nargeot, Romuald; Jelescu, Ileana Ozana; Le Bihan, Denis; Ciobanu, Luisa

    2014-01-01

    In this work, we show the feasibility of performing functional MRI studies with single-cell resolution. At ultrahigh magnetic field, manganese-enhanced magnetic resonance microscopy allows the identification of most motor neurons in the buccal network of Aplysia at low, nontoxic Mn2+ concentrations. We establish that Mn2+ accumulates intracellularly on injection into the living Aplysia and that its concentration increases when the animals are presented with a sensory stimulus. We also show that we can distinguish between neuronal activities elicited by different types of stimuli. This method opens up a new avenue into probing the functional organization and plasticity of neuronal networks involved in goal-directed behaviors with single-cell resolution. PMID:24872449

  15. Nanoscale capacitance imaging with attofarad resolution using ac current sensing atomic force microscopy

    International Nuclear Information System (INIS)

    Fumagalli, L; Ferrari, G; Sampietro, M; Casuso, I; MartInez, E; Samitier, J; Gomila, G

    2006-01-01

    Nanoscale capacitance imaging with attofarad resolution (∼1 aF) of a nano-structured oxide thin film, using ac current sensing atomic force microscopy, is reported. Capacitance images are shown to follow the topographic profile of the oxide closely, with nanometre vertical resolution. A comparison between experimental data and theoretical models shows that the capacitance variations observed in the measurements can be mainly associated with the capacitance probed by the tip apex and not with positional changes of stray capacitance contributions. Capacitance versus distance measurements further support this conclusion. The application of this technique to the characterization of samples with non-voltage-dependent capacitance, such as very thin dielectric films, self-assembled monolayers and biological membranes, can provide new insight into the dielectric properties at the nanoscale

  16. Hard X-ray Microscopy with sub 30 nm Spatial Resolution

    International Nuclear Information System (INIS)

    Tang, M.-T.; Song, Y.-F.; Yin, G.-C.; Chen, J.-H.; Chen, Y.-M.; Liang, Keng S.; Chen, F.-R.; Duewer, F.; Yun Wenbing

    2007-01-01

    A transmission X-ray microscope (TXM) has been installed at the BL01B beamline at National Synchrotron Radiation Research Center in Taiwan. This state-of-the-art TXM operational in a range 8-11 keV provides 2D images and 3D tomography with spatial resolution 60 nm, and with the Zernike-phase contrast mode for imaging light materials such as biological specimens. A spatial resolution of the TXM better than 30 nm, apparently the best result in hard X-ray microscopy, has been achieved by employing the third diffraction order of the objective zone plate. The TXM has been applied in diverse research fields, including analysis of failure mechanisms in microelectronic devices, tomographic structures of naturally grown photonic specimens, and the internal structure of fault zone gouges from an earthquake core. Here we discuss the scope and prospects of the project, and the progress of the TXM in NSRRC

  17. Nanoscale capacitance imaging with attofarad resolution using ac current sensing atomic force microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Fumagalli, L [Dipartimento di Elettronica e Informazione, Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 (Italy); Ferrari, G [Dipartimento di Elettronica e Informazione, Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 (Italy); Sampietro, M [Dipartimento di Elettronica e Informazione, Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 (Italy); Casuso, I [Departament d' Electronica, Universitat de Barcelona, C/MartIi Franques 1, 08028 Barcelona (Spain); MartInez, E [Plataforma de Nanotecnologia, Parc Cientific de Barcelona, C/ Josep Samitier 1-5, 08028-Barcelona (Spain); Samitier, J [Departament d' Electronica, Universitat de Barcelona, C/MartIi Franques 1, 08028 Barcelona (Spain); Gomila, G [Departament d' Electronica, Universitat de Barcelona, C/MartIi Franques 1, 08028 Barcelona (Spain)

    2006-09-28

    Nanoscale capacitance imaging with attofarad resolution ({approx}1 aF) of a nano-structured oxide thin film, using ac current sensing atomic force microscopy, is reported. Capacitance images are shown to follow the topographic profile of the oxide closely, with nanometre vertical resolution. A comparison between experimental data and theoretical models shows that the capacitance variations observed in the measurements can be mainly associated with the capacitance probed by the tip apex and not with positional changes of stray capacitance contributions. Capacitance versus distance measurements further support this conclusion. The application of this technique to the characterization of samples with non-voltage-dependent capacitance, such as very thin dielectric films, self-assembled monolayers and biological membranes, can provide new insight into the dielectric properties at the nanoscale.

  18. Helium Ion Microscopy (HIM) for the imaging of biological samples at sub-nanometer resolution

    Science.gov (United States)

    Joens, Matthew S.; Huynh, Chuong; Kasuboski, James M.; Ferranti, David; Sigal, Yury J.; Zeitvogel, Fabian; Obst, Martin; Burkhardt, Claus J.; Curran, Kevin P.; Chalasani, Sreekanth H.; Stern, Lewis A.; Goetze, Bernhard; Fitzpatrick, James A. J.

    2013-12-01

    Scanning Electron Microscopy (SEM) has long been the standard in imaging the sub-micrometer surface ultrastructure of both hard and soft materials. In the case of biological samples, it has provided great insights into their physical architecture. However, three of the fundamental challenges in the SEM imaging of soft materials are that of limited imaging resolution at high magnification, charging caused by the insulating properties of most biological samples and the loss of subtle surface features by heavy metal coating. These challenges have recently been overcome with the development of the Helium Ion Microscope (HIM), which boasts advances in charge reduction, minimized sample damage, high surface contrast without the need for metal coating, increased depth of field, and 5 angstrom imaging resolution. We demonstrate the advantages of HIM for imaging biological surfaces as well as compare and contrast the effects of sample preparation techniques and their consequences on sub-nanometer ultrastructure.

  19. Concept for room temperature single-spin tunneling force microscopy with atomic spatial resolution

    Science.gov (United States)

    Payne, Adam

    A study of a force detected single-spin magnetic resonance measurement concept with atomic spatial resolution is presented. The method is based upon electrostatic force detection of spin-selection rule controlled single electron tunneling between two electrically isolated paramagnetic states. Single-spin magnetic resonance detection is possible by measuring the force detected tunneling charge noise on and off spin resonance. Simulation results of this charge noise, based upon physical models of the tunneling and spin physics, are directly compared to measured atomic force microscopy (AFM) system noise. The results show that the approach could provide single-spin measurement of electrically isolated defect states with atomic spatial resolution at room temperature.

  20. Atomic-resolution single-spin magnetic resonance detection concept based on tunneling force microscopy

    Science.gov (United States)

    Payne, A.; Ambal, K.; Boehme, C.; Williams, C. C.

    2015-05-01

    A study of a force detected single-spin magnetic resonance measurement concept with atomic spatial resolution is presented. The method is based upon electrostatic force detection of spin-selection rule controlled single-electron tunneling between two electrically isolated paramagnetic states. Single-spin magnetic resonance detection is possible by measuring the force detected tunneling charge noise on and off spin resonance. Simulation results of this charge noise, based upon physical models of the tunneling and spin physics, are directly compared to measured atomic force microscopy system noise. The results show that the approach could provide single-spin measurement of electrically isolated qubit states with atomic spatial resolution at room temperature.

  1. SUPER-RESOLUTION ULTRASOUND TOMOGRAPHY: A PRELIMINARY STUDY WITH A RING ARRAY

    Energy Technology Data Exchange (ETDEWEB)

    HUANG, LIANJIE [Los Alamos National Laboratory; SIMONETTI, FRANCESCO [NON LANL; DURIC, NEBOJSA [NON LANL; RAMA, OLSI [NON LANL

    2007-01-18

    Ultrasound tomography attempts to retrieve the structure of an objective by exploiting the interaction of acoustic waves with the object. A fundamental limit of ultrasound tomography is that features cannot be resolved if they are spaced less than {lambda}/2 apart, where {lambda} is wavelength of the probing wave, regardless of the degree of accuracy of the measurements. Therefore, since the attenuation of the probing wave with propagation distance increases as {lambda} decreases, resolution has to be traded against imaging depth. Recently, it has been shown that the {lambda}/2 limit is a consequence of the Born approximation (implicit in the imaging algorithms currently employed) which neglects the distortion of the probing wavefield as it travels through the medium to be imaged. On the other hand, such a distortion, which is due to the multiple scattering phenomenon, can encode unlimited resolution in the radiating component of the scattered field. Previously, a resolution better than {lambda}/3 has been reported in these proceedings [F. Simonetti, pp. 126 (2006)] in the case of elastic wave probing. In this paper, they demonstrate experimentally a resolution better than {lambda}/4 for objects immersed in a water bth probed by means of a ring array which excites and detects pressure waves in a full view configuration.

  2. Dark-field hyperlens: Super-resolution imaging of weakly scattering objects

    DEFF Research Database (Denmark)

    Repän, Taavi; Lavrinenko, Andrei; Zhukovsky, Sergei

    2015-01-01

    : We propose a device for subwavelength optical imaging based on a metal-dielectric multilayer hyperlens designed in such a way that only large-wavevector (evanescent) waves are transmitted while all propagating (small-wavevector) waves from the object area are blocked by the hyperlens. We...... numerically demonstrate that as the result of such filtering, the image plane only contains scattered light from subwavelength features of the objects and is completely free from background illumination. Similar in spirit to conventional dark-field microscopy, the proposed dark-field hyperlens is shown...

  3. Convex relaxations of spectral sparsity for robust super-resolution and line spectrum estimation

    Science.gov (United States)

    Chi, Yuejie

    2017-08-01

    We consider recovering the amplitudes and locations of spikes in a point source signal from its low-pass spectrum that may suffer from missing data and arbitrary outliers. We first review and provide a unified view of several recently proposed convex relaxations that characterize and capitalize the spectral sparsity of the point source signal without discretization under the framework of atomic norms. Next we propose a new algorithm when the spikes are known a priori to be positive, motivated by applications such as neural spike sorting and fluorescence microscopy imaging. Numerical experiments are provided to demonstrate the effectiveness of the proposed approach.

  4. Instant live-cell super-resolution imaging of cellular structures by nanoinjection of fluorescent probes.

    Science.gov (United States)

    Hennig, Simon; van de Linde, Sebastian; Lummer, Martina; Simonis, Matthias; Huser, Thomas; Sauer, Markus

    2015-02-11

    Labeling internal structures within living cells with standard fluorescent probes is a challenging problem. Here, we introduce a novel intracellular staining method that enables us to carefully control the labeling process and provides instant access to the inner structures of living cells. Using a hollow glass capillary with a diameter of <100 nm, we deliver functionalized fluorescent probes directly into the cells by (di)electrophoretic forces. The label density can be adjusted and traced directly during the staining process by fluorescence microscopy. We demonstrate the potential of this technique by delivering and imaging a range of commercially available cell-permeable and nonpermeable fluorescent probes to cells.

  5. Portable optical-resolution photoacoustic microscopy for volumetric imaging of multiscale organisms.

    Science.gov (United States)

    Jin, Tian; Guo, Heng; Yao, Lei; Xie, Huikai; Jiang, Huabei; Xi, Lei

    2018-04-01

    Photoacoustic microscopy (PAM) provides a fundamentally new tool for a broad range of studies of biological structures and functions. However, the use of PAM has been largely limited to small vertebrates due to the large size/weight and the inconvenience of the equipment. Here, we describe a portable optical-resolution photoacoustic microscopy (pORPAM) system for 3-dimensional (3D) imaging of small-to-large rodents and humans with a high spatiotemporal resolution and a large field of view. We show extensive applications of pORPAM to multiscale animals including mice and rabbits. In addition, we image the 3D vascular networks of human lips, and demonstrate the feasibility of pORPAM to observe the recovery process of oral ulcer and cancer-associated capillary loops in human oral cavities. This technology is promising for broad biomedical studies from fundamental biology to clinical diseases. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Single image super-resolution using locally adaptive multiple linear regression.

    Science.gov (United States)

    Yu, Soohwan; Kang, Wonseok; Ko, Seungyong; Paik, Joonki

    2015-12-01

    This paper presents a regularized superresolution (SR) reconstruction method using locally adaptive multiple linear regression to overcome the limitation of spatial resolution of digital images. In order to make the SR problem better-posed, the proposed method incorporates the locally adaptive multiple linear regression into the regularization process as a local prior. The local regularization prior assumes that the target high-resolution (HR) pixel is generated by a linear combination of similar pixels in differently scaled patches and optimum weight parameters. In addition, we adapt a modified version of the nonlocal means filter as a smoothness prior to utilize the patch redundancy. Experimental results show that the proposed algorithm better restores HR images than existing state-of-the-art methods in the sense of the most objective measures in the literature.

  7. High-resolution MRI using a microscopy coil for the diagnosis of recurrent lateral patellar dislocation

    International Nuclear Information System (INIS)

    Iwata, Yuki; Fujii, Masahiko; Muratsu, Hirotsugu; Kurosaka, Masahiro; Kawamitsu, Hideaki; Sugimura, Kazuro; Shibanuma, Hitoshi

    2006-01-01

    Magnetic resonance imaging (MRI) has been commonly used for the preoperative evaluation of recurrent lateral patellar dislocation (RLPD). The purpose of this study was to determine the usefulness of high-resolution MRI (HR-MRI) with a microscopy coil for diagnosing RLPD. The study group consisted of 15 patients with clinically diagnosed RLPD and 10 normal volunteers. All studies were performed on a 1.5-T MR system. First, conventional MRIs of the whole knee joint were obtained using the knee coil. Then HR-MRI scans using a microscopy coil in the medial aspect of the patella were obtained at the level of the superior pole of the patella, targeting the medial patellofemoral ligament (MPFL). The acquired HR-MRIs with RLPD were reviewed concerning the MPFL injury and the patellar injury. The MPFL was distinguished as a separate ligament, and the layer structure of the patellar cartilage was visualized clearly in all volunteers. The MPFL injury was visualized in 12 cases (87%); it included discontinuity, thickening, and loosening. The patellar injury was visualized in 11 cases (73%), which included dissecans of the medial margin and cartilage injuries. HR-MRI with a microscopy coil provides precise information of the MPFL and patellar cartilage injury for the diagnosis of RLPD. (author)

  8. Microscopy

    Science.gov (United States)

    Patricia A. Moss; Les Groom

    2001-01-01

    Microscopy is the study and interpretation of images produced by a microscope. "Interpretation" is the keyword, because the microscope enables one to see structures that are too small or too close together to be resolved by the unaided eye. (The human eye cannot separate two points or lines that are closer together than 0.1 mm.) it is important to...

  9. Atomic-resolution environmental TEM for quantitative in-situ microscopy in materials science.

    Science.gov (United States)

    Takeda, Seiji; Yoshida, Hideto

    2013-02-01

    We have compiled our recent in-situ quantitative environmental transmission electron microscopy (ETEM) studies on typical gold nanoparticulate catalysts for the low-temperature oxidation of CO to describe the issues surrounding the application of ETEM, with a special regard to catalyst chemistry. Thanks to the recent development of high-resolution environmental transmission electron microscopes that can work robustly to accumulate observation data in controlled environments, we can deal with the electron irradiation effects and heterogeneity of real catalysts. We established a structural evolution diagram that summarizes the structure of catalysts under electron irradiation as a function of the electron current density ϕ and the electron dose, D. By extrapolating to ϕ = 0, D = 0, we could deduce the intrinsic catalysis structure (without electron irradiation) in various environments, including reaction environments. By numerically and statistically analyzing a substantial number of ETEM images of gold nanoparticles, we established a morphology phase diagram that summarizes how the majority of gold nanoparticles change their morphology systematically as a function of the partial pressures of CO and O(2). Similar diagrams will be helpful in elucidating the phenomena that directly correlate with the catalytic activity determined from ETEM observations. Using these quantitative analyses, we could analyze Cs-corrected ETEM images of the catalysts. The surfaces of gold nanoparticles were structurally reconstructed under reaction conditions, via interactions with CO molecules. CO molecules were observed on the surfaces of catalysts under reaction conditions using high-resolution ETEM. Finally, we discuss the potential of environmental transmission electron microscopy for quantitative in-situ microscopy at the atomic scale.

  10. High resolution Transmission Electron Microscopy characterization of a milled oxide dispersion strengthened steel powder

    Energy Technology Data Exchange (ETDEWEB)

    Loyer-Prost, M., E-mail: marie.loyer-prost@cea.fr [DEN-Service de Recherches de Métallurgie Physique, CEA, Université Paris-Saclay, F-91191, Gif-sur-Yvette (France); Merot, J.-S. [Laboratoire d’Etudes des Microstructures – UMR 104, CNRS/ONERA, BP72-29, Avenue de la Division Leclerc, 92 322, Châtillon (France); Ribis, J. [DEN-Service de Recherches de Métallurgie Appliquée, CEA, Université Paris-Saclay, F-91191, Gif-sur-Yvette (France); Le Bouar, Y. [Laboratoire d’Etudes des Microstructures – UMR 104, CNRS/ONERA, BP72-29, Avenue de la Division Leclerc, 92 322, Châtillon (France); Chaffron, L. [DEN-Service de Recherches de Métallurgie Appliquée, CEA, Université Paris-Saclay, F-91191, Gif-sur-Yvette (France); Legendre, F. [DEN-Service de la Corrosion et du Comportement des Matériaux dans leur Environnement, CEA, Université Paris-Saclay, F-91191, Gif-sur-Yvette (France)

    2016-10-15

    Oxide Dispersion Strengthened (ODS) steels are promising materials for generation IV fuel claddings as their dense nano-oxide dispersion provides good creep and irradiation resistance. Even if they have been studied for years, the formation mechanism of these nano-oxides is still unclear. Here we report for the first time a High Resolution Transmission Electron Microscopy and Energy Filtered Transmission Electron Microscopy characterization of an ODS milled powder. It provides clear evidence of the presence of small crystalline nanoclusters (NCs) enriched in titanium directly after milling. Small NCs (<5 nm) have a crystalline structure and seem partly coherent with the matrix. They have an interplanar spacing close to the (011) {sub bcc} iron structure. They coexist with larger crystalline spherical precipitates of 15–20 nm in size. Their crystalline structure may be metastable as they are not consistent with any Y-Ti-O or Ti-O structure. Such detailed observations in the as-milled grain powder confirm a mechanism of Y, Ti, O dissolution in the ferritic matrix followed by a NC precipitation during the mechanical alloying process of ODS materials. - Highlights: • We observed an ODS ball-milled powder by high resolution transmission microscopy. • The ODS ball-milled powder exhibits a lamellar microstructure. • Small crystalline nanoclusters were detected in the milled ODS powder. • The nanoclusters in the ODS milled powder are enriched in titanium. • Larger NCs of 15–20 nm in size are, at least, partly coherent with the matrix.

  11. High-Resolution 3 T MR Microscopy Imaging of Arterial Walls

    International Nuclear Information System (INIS)

    Sailer, Johannes; Rand, Thomas; Berg, Andreas; Sulzbacher, Irene; Peloschek, P.; Hoelzenbein, Thomas; Lammer, Johannes

    2006-01-01

    Purpose. To achieve a high spatial resolution in MR imaging that allows for clear visualization of anatomy and even histology and documentation of plaque morphology in in vitro samples from patients with advanced atherosclerosis. A further objective of our study was to evaluate whether T2-weighted high-resolution MR imaging can provide accurate classification of atherosclerotic plaque according to a modified American Heart Association classification. Methods. T2-weighted images of arteries were obtained in 13 in vitro specimens using a 3 T MR unit (Medspec 300 Avance/Bruker, Ettlingen, Germany) combined with a dedicated MR microscopy system. Measurement parameters were: T2-weighted sequences with TR 3.5 sec, TE 15-120 msec; field of view (FOV) 1.4 x 1.4; NEX 8; matrix 192; and slice thickness 600 μm. MR measurements were compared with corresponding histologic sections. Results. We achieved excellent spatial and contrast resolution in all specimens. We found high agreement between MR images and histology with regard to the morphology and extent of intimal proliferations in all but 2 specimens. We could differentiate fibrous caps and calcifications from lipid plaque components based on differences in signal intensity in order to differentiate hard and soft atheromatous plaques. Hard plaques with predominantly intimal calcifications were found in 7 specimens, and soft plaques with a cholesterol/lipid content in 5 cases. In all specimens, hemorrhage or thrombus formation, and fibrotic and hyalinized tissue could be detected on both MR imaging and histopathology. Conclusion. High-resolution, high-field MR imaging of arterial walls demonstrates the morphologic features, volume, and extent of intimal proliferations with high spatial and contrast resolution in in vitro specimens and can differentiate hard and soft plaques

  12. Photonic nanojet super-resolution in immersed ordered assembly of dielectric microspheres

    International Nuclear Information System (INIS)

    Geints, Y.E.; Zemlyanov, A.A.

    2017-01-01

    Highlights: • Ordered microassemblies of dielectric microspheres immersed in a transparent matrix exposed to a light wave produce an array of the “photonic nanojets” coupled through the field interference. • “Photonic nanojet” parameters depend on the inter-sphere gap and immersion depth. • Sphere microassembly can produce the jets with the sub-diffraction spatial resolution if the particles are loosely packed and semi-immersed. - Abstract: Specific spatially-localized optical field structure, which is often referred to as a photonic nanojet (PNJ), is formed in the near-field scattering area of non-absorbing dielectric micron-sized particle exposed to an optical radiation. By virtue of the finite-difference time-domain technique we numerically simulate the two-dimensional array of PNJs created by an ordered single-layer microassembly of glass microspheres immersed in a transparent polymer matrix. The behavior of the main PNJ parameters (length, diameter, and intensity) is analyzed subject to the immersion depth of the microparticles and cooperative interference effects of the neighboring microspheres. We show that depending on microassembly configuration, the PNJ quality can be significantly improved; in particular, the PNJ spatial resolution better than λ/5 can be achieved.

  13. Practical Approaches to Mitigation of Specimen Charging in High-Resolution Transmission Electron Microscopy

    Directory of Open Access Journals (Sweden)

    Young-Min Kim

    2010-09-01

    Full Text Available Specimen charging that is associated with the electron bombardment on the sample is a practical hindrance to high-resolution transmission electron microscopy (HRTEM analysis because it causes a severe loss of resolution in either diffraction or image data. Conductive thin film deposition on an insulating specimen has been proposed as an effective approach to the mitigation of the specimen charging; however, this method is generally not useful in HRTEM imaging of materials because the deposited film induces another artifact in the HRTEM image contrast. In this study, we propose practical methods to mitigate the specimen charging that takes place during the HRTEM of materials. For bulk-type specimens prepared by either an ion-thinning or focused-ion beam (FIB process, a plasma cleaning treatment is significantly effective in eliminating the charging phenomenon. In the case of low-dimensional nanomaterials such as nanowires and nanoparticles, the plasma cleaning is not feasible; however, the charging effect can be effectively eliminated by adjusting the electron illumination condition. The proposed methods facilitate a decrease in the buildup of specimen charging, thereby enhancing the quality of high-resolution images significantly.

  14. High-time resolution conjugate SuperDARN radar observations of the dayside convection response to changes in IMF By

    Directory of Open Access Journals (Sweden)

    G. Chisham

    2000-02-01

    Full Text Available We present data from conjugate SuperDARN radars describing the high-latitude ionosphere's response to changes in the direction of IMF By during a period of steady IMF Bz southward and Bx positive. During this interval, the radars were operating in a special mode which gave high-time resolution data (30 s sampling period on three adjacent beams with a full scan every 3 min. The location of the radars around magnetic local noon at the time of the event allowed detailed observations of the variations in the ionospheric convection patterns close to the cusp region as IMF By varied. A significant time delay was observed in the ionospheric response to the IMF By changes between the two hemispheres. This is explained as being partially a consequence of the location of the dominant merging region on the magnetopause, which is ~8-12RE closer to the northern ionosphere than to the southern ionosphere (along the magnetic field line due to the dipole tilt of the magnetosphere and the orientation of the IMF. This interpretation supports the anti-parallel merging hypothesis and highlights the importance of the IMF Bx component in solar wind-magnetosphere coupling.Key words: Ionosphere (plasma convection - Magnetospheric physics (magnetopause, cusp, and boundary layers; solar wind - magnetosphere interactions

  15. Reaction Control System Thruster Cracking Consultation: NASA Engineering and Safety Center (NESC) Materials Super Problem Resolution Team (SPRT) Findings

    Science.gov (United States)

    MacKay, Rebecca A.; Smith, Stephen W.; Shah, Sandeep R.; Piascik, Robert S.

    2005-01-01

    The shuttle orbiter s reaction control system (RCS) primary thruster serial number 120 was found to contain cracks in the counter bores and relief radius after a chamber repair and rejuvenation was performed in April 2004. Relief radius cracking had been observed in the 1970s and 1980s in seven thrusters prior to flight; however, counter bore cracking had never been seen previously in RCS thrusters. Members of the Materials Super Problem Resolution Team (SPRT) of the NASA Engineering and Safety Center (NESC) conducted a detailed review of the relevant literature and of the documentation from the previous RCS thruster failure analyses. It was concluded that the previous failure analyses lacked sufficient documentation to support the conclusions that stress corrosion cracking or hot-salt cracking was the root cause of the thruster cracking and lacked reliable inspection controls to prevent cracked thrusters from entering the fleet. The NESC team identified and performed new materials characterization and mechanical tests. It was determined that the thruster intergranular cracking was due to hydrogen embrittlement and that the cracking was produced during manufacturing as a result of processing the thrusters with fluoride-containing acids. Testing and characterization demonstrated that appreciable environmental crack propagation does not occur after manufacturing.

  16. Synthesis of a Far‐Red Photoactivatable Silicon‐Containing Rhodamine for Super‐Resolution Microscopy

    Science.gov (United States)

    Grimm, Jonathan B.; Klein, Teresa; Kopek, Benjamin G.; Shtengel, Gleb; Hess, Harald F.; Sauer, Markus

    2015-01-01

    Abstract The rhodamine system is a flexible framework for building small‐molecule fluorescent probes. Changing N‐substitution patterns and replacing the xanthene oxygen with a dimethylsilicon moiety can shift the absorption and fluorescence emission maxima of rhodamine dyes to longer wavelengths. Acylation of the rhodamine nitrogen atoms forces the molecule to adopt a nonfluorescent lactone form, providing a convenient method to make fluorogenic compounds. Herein, we take advantage of all of these structural manipulations and describe a novel photoactivatable fluorophore based on a Si‐containing analogue of Q‐rhodamine. This probe is the first example of a “caged” Si‐rhodamine, exhibits higher photon counts compared to established localization microscopy dyes, and is sufficiently red‐shifted to allow multicolor imaging. The dye is a useful label for super‐resolution imaging and constitutes a new scaffold for far‐red fluorogenic molecules. PMID:26661345

  17. Nano features of Al/Au ultrasonic bond interface observed by high resolution transmission electron microscopy

    International Nuclear Information System (INIS)

    Ji Hongjun; Li Mingyu; Kim, Jong-Myung; Kim, Dae-Won; Wang Chunqing

    2008-01-01

    Nano-scale interfacial details of ultrasonic AlSi1 wire wedge bonding to a Au/Ni/Cu pad were investigated using high resolution transmission electron microscopy (HRTEM). The intermetallic phase Au 8 Al 3 formed locally due to diffusion and reaction activated by ultrasound at the Al/Au bond interface. Multilayer sub-interfaces roughly parallel to the wire/pad interface were observed among this phase, and interdiffusional features near the Au pad resembled interference patterns, alternately dark and bright bars. Solid-state diffusion theory cannot be used to explain why such a thick compound formed within milliseconds at room temperature. The major formation of metallurgical bonds was attributed to ultrasonic cyclic vibration

  18. High-resolution TEM microscopy study of the creep behaviour of carbon-based cathode materials

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Wei, E-mail: wwlyzwkj@126.com [College of Materials Science and Engineering, Henan University of Science and Technology, Luoyang 471023 (China); Collaborative Innovation Center of Nonferrous Metals Henan Province, Luoyang 471023 (China); Chen, Weijie [College of Materials Science and Engineering, Henan University of Science and Technology, Luoyang 471023 (China); Collaborative Innovation Center of Nonferrous Metals Henan Province, Luoyang 471023 (China); Gu, Wanduo [Collaborative Innovation Center of Nonferrous Metals Henan Province, Luoyang 471023 (China)

    2017-02-27

    Creep is in close relationship with the materials deterioration and deformation of the cathodes in aluminum reduction cells. The purpose of this work is to obtain the creep mechanism of the carbon cathode for aluminum electrolysis. A modified Rapoport equipment was used for measuring the creep strain of the semi-graphitic cathodes during aluminum electrolysis with CR=2.5 and at temperature of 945 ℃. The arrangement of carbon atom has been studied after hexagonal graphite converting into rhombohedral graphite during aluminum electrolysis by XRD and high-resolution transmission electron microscopy (HRTEM). The creep deformation of the carbon cathode has a close relationship with the mobile dislocation walls. These results will be helpful in controlling the cathode quality and its performance in aluminum reduction cells.

  19. High resolution electron microscopy study of as-prepared and annealed tungsten-carbon multilayers

    International Nuclear Information System (INIS)

    Nguyen, T.D.; Gronsky, R.; Kortright, J.B.

    1988-12-01

    A series of sputtered tungsten-carbon multilayer structures with periods ranging from 2 to 12 nm in the as-prepared state and after annealing at 500/degree/C for 4 hours has been studied with high resolution transmission electron microscopy. The evolution with annealing of the microstructure of these multilayers depends on their period. As-prepared structures appear predominantly amorphous from TEM imaging and diffraction. Annealing results in crystallization of the W-rich layers into WC in the larger period samples, and less complete or no crystallization in the smaller period samples. X-ray scattering reveals that annealing expands the period in a systematic way. The layers remain remarkably well-defined after annealing under these conditions. 12 refs., 4 figs., 1 tab

  20. High-resolution transmission electron microscopy of grain-refining particles in amorphous aluminum alloys

    International Nuclear Information System (INIS)

    Schumacher, P.; Greer, A.L.

    1996-01-01

    The nucleation mechanism of Al-Ti-B grain refiners is studied in an Al-based amorphous alloy. The ability to limit growth of α-Al in the amorphous alloy permits the microscopical observation of nucleation events on boride particles. Earlier studies of this kind are extended by using high-resolution electron microscopy. This shows that the efficient nucleation α-Al depends on the TiB 2 particles being coated with a thin layer of Al 3 Ti, which can form only when there is some excess titanium in the melt. The aluminide layer, stabilized by adsorption effects, can be as little as a few monolayers thick, and is coherent with the boride. The nature of this layer, and its importance for the nucleation mechanism are discussed. The fading of the grain refinement action is also considered

  1. Characterisation of phases in nanostructured, multilayered titanium alloys by analytical and high-resolution electron microscopy.

    Science.gov (United States)

    Czyrska-Filemonowicz, A; Buffat, P A

    2009-01-01

    Surface processing of a Ti-6Al-4V alloy led to a complex multilayered microstructure containing several phases of the Ni-Ti-P-Al-O system, which improves the mechanical and tribological surface properties. The microstructure, chemical and phase compositions of the hard layer formed on the surface were investigated by LM, XRD, SEM as well as analytical/high-resolution TEM, STEM, EDS, electron diffraction and FIB. Phase identification based on electron diffraction, HRTEM and EDS microanalysis revealed the presence of several binary and ternary phases in the system Ti-Ni-P, sometimes with partial substitution of Ti by Al. However some phases, mainly nanoparticles, still remain not identified satisfactorily. Electron microscopy techniques used for identification of phases present in surface multilayers and some practical limits to their routine application are reminded here.

  2. Study on super-resolution three-dimensional range-gated imaging technology

    Science.gov (United States)

    Guo, Huichao; Sun, Huayan; Wang, Shuai; Fan, Youchen; Li, Yuanmiao

    2018-04-01

    Range-gated three dimensional imaging technology is a hotspot in recent years, because of the advantages of high spatial resolution, high range accuracy, long range, and simultaneous reflection of target reflectivity information. Based on the study of the principle of intensity-related method, this paper has carried out theoretical analysis and experimental research. The experimental system adopts the high power pulsed semiconductor laser as light source, gated ICCD as the imaging device, can realize the imaging depth and distance flexible adjustment to achieve different work mode. The imaging experiment of small imaging depth is carried out aiming at building 500m away, and 26 group images were obtained with distance step 1.5m. In this paper, the calculation method of 3D point cloud based on triangle method is analyzed, and 15m depth slice of the target 3D point cloud are obtained by using two frame images, the distance precision is better than 0.5m. The influence of signal to noise ratio, illumination uniformity and image brightness on distance accuracy are analyzed. Based on the comparison with the time-slicing method, a method for improving the linearity of point cloud is proposed.

  3. Evaluation of Composite Wire Ropes Using Unsaturated Magnetic Excitation and Reconstruction Image with Super-Resolution

    Directory of Open Access Journals (Sweden)

    Xiaojiang Tan

    2018-05-01

    Full Text Available Estimating the exact residual lifetime of wire rope involves the security of industry manufacturing, mining, tourism, and so on. In this paper, a novel testing technology was developed based on unsaturated magnetic excitation, and a fabricating prototype overcame the shortcomings of traditional detection equipment in terms of volume, sensibility, reliability, and weight. Massive artificial discontinuities were applied to examine the effectiveness of this new technology with a giant magneto resistance(GMR sensor array, which included types of small gaps, curling wires, wide fractures, and abrasion. A resolution enhancement method, which was adopted for multiframe images, was proposed for promoting magnetic flux leakage images of a few sensors. Characteristic vectors of statistics and geometry were extracted, then we applied a radial basis function neural network to achieve a quantitative recognition rate of 91.43% with one wire-limiting error. Experimental results showed that the new device can detect defects in various types of wire rope and prolong the service life with high lift-off distance and high reliability, and the system could provide useful options to evaluate the lifetime of wire rope.

  4. X-ray microscopy with high resolution zone plates -- Recent developments

    International Nuclear Information System (INIS)

    Schneider, G.; Wilhein, T.; Niemann, B.; Guttmann, P.

    1995-01-01

    In order to expand the applications of X-ray microscopy, developments in the fields of zone plate technology, specimen preparation and imaging techniques have been made. A new cross-linked polymer chain electron beam resist allows to record zone plate pattern down to 19 nm outermost zone width. High resolution zone plates in germanium with outermost zone widths down to 19 nm have been developed. In addition, phase zone plates in nickel down to 30 nm zone width have been made by electroplating. In order to enhance the image contrast for weak absorbing objects, the phase contrast method for X-ray microscopy was developed and implemented on the Goettingen X-ray microscope at BESSY. The effects of X-ray absorption on the structure of biological specimen limits the maximum applicable radiation dose and therefore the achievable signal to noise ratio for an artifact-free X-ray image. To improve the stability especially of biological specimen, a cryogenic object chamber has been developed and tested. It turns out that at the operating temperature T ≤ 130 K unfixed biological specimen can be exposed to a radiation dose of 10 9 --10 10 Gy without any observable structural changes. A multiple-angle viewing stage allows to take stereoscopic images with the X-ray microscope, giving a 3D-impression of the object

  5. Helium ion microscopy and ultra-high-resolution scanning electron microscopy analysis of membrane-extracted cells reveals novel characteristics of the cytoskeleton of Giardia intestinalis.

    Science.gov (United States)

    Gadelha, Ana Paula Rocha; Benchimol, Marlene; de Souza, Wanderley

    2015-06-01

    Giardia intestinalis presents a complex microtubular cytoskeleton formed by specialized structures, such as the adhesive disk, four pairs of flagella, the funis and the median body. The ultrastructural organization of the Giardia cytoskeleton has been analyzed using different microscopic techniques, including high-resolution scanning electron microscopy. Recent advances in scanning microscopy technology have opened a new venue for the characterization of cellular structures and include scanning probe microscopy techniques such as ultra-high-resolution scanning electron microscopy (UHRSEM) and helium ion microscopy (HIM). Here, we studied the organization of the cytoskeleton of G. intestinalis trophozoites using UHRSEM and HIM in membrane-extracted cells. The results revealed a number of new cytoskeletal elements associated with the lateral crest and the dorsal surface of the parasite. The fine structure of the banded collar was also observed. The marginal plates were seen linked to a network of filaments, which were continuous with filaments parallel to the main cell axis. Cytoplasmic filaments that supported the internal structures were seen by the first time. Using anti-actin antibody, we observed a labeling in these filamentous structures. Taken together, these data revealed new surface characteristics of the cytoskeleton of G. intestinalis and may contribute to an improved understanding of the structural organization of trophozoites. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Electron beam fabrication and characterization of high-resolution magnetic force microscopy tips

    Science.gov (United States)

    Rührig, M.; Porthun, S.; Lodder, J. C.; McVitie, S.; Heyderman, L. J.; Johnston, A. B.; Chapman, J. N.

    1996-03-01

    The stray field, magnetic microstructure, and switching behavior of high-resolution electron beam fabricated thin film tips for magnetic force microscopy (MFM) are investigated with different imaging modes in a transmission electron microscope (TEM). As the tiny smooth carbon needles covered with a thermally evaporated magnetic thin film are transparent to the electron energies used in these TEMs it is possible to observe both the external stray field emanating from the tips as well as their internal domain structure. The experiments confirm the basic features of electron beam fabricated thin film tips concluded from various MFM observations using these tips. Only a weak but highly concentrated stray field is observed emanating from the immediate apex region of the tip, consistent with their capability for high resolution. It also supports the negligible perturbation of the magnetization sample due to the tip stray field observed in MFM experiments. Investigation of the magnetization distributions within the tips, as well as preliminary magnetizing experiments, confirm a preferred single domain state of the high aspect ratio tips. To exclude artefacts of the observation techniques both nonmagnetic tips and those supporting different magnetization states are used for comparison.

  7. DESIGN OF DYADIC-INTEGER-COEFFICIENTS BASED BI-ORTHOGONAL WAVELET FILTERS FOR IMAGE SUPER-RESOLUTION USING SUB-PIXEL IMAGE REGISTRATION

    Directory of Open Access Journals (Sweden)

    P.B. Chopade

    2014-05-01

    Full Text Available This paper presents image super-resolution scheme based on sub-pixel image registration by the design of a specific class of dyadic-integer-coefficient based wavelet filters derived from the construction of a half-band polynomial. First, the integer-coefficient based half-band polynomial is designed by the splitting approach. Next, this designed half-band polynomial is factorized and assigned specific number of vanishing moments and roots to obtain the dyadic-integer coefficients low-pass analysis and synthesis filters. The possibility of these dyadic-integer coefficients based wavelet filters is explored in the field of image super-resolution using sub-pixel image registration. The two-resolution frames are registered at a specific shift from one another to restore the resolution lost by CCD array of camera. The discrete wavelet transform (DWT obtained from the designed coefficients is applied on these two low-resolution images to obtain the high resolution image. The developed approach is validated by comparing the quality metrics with existing filter banks.

  8. TH-EF-BRA-11: Feasibility of Super-Resolution Time-Resolved 4DMRI for Multi-Breath Volumetric Motion Simulation in Radiotherapy Planning

    Energy Technology Data Exchange (ETDEWEB)

    Li, G; Zakian, K; Deasy, J [Memorial Sloan Kettering Cancer Center, New York, NY (United States); Wei, J [City College of New York, New York, NY (United States); Hunt, M [Mem Sloan-Kettering Cancer Ctr, New York, NY (United States)

    2016-06-15

    Purpose: To develop a novel super-resolution time-resolved 4DMRI technique to evaluate multi-breath, irregular and complex organ motion without respiratory surrogate for radiotherapy planning. Methods: The super-resolution time-resolved (TR) 4DMRI approach combines a series of low-resolution 3D cine MRI images acquired during free breathing (FB) with a high-resolution breath-hold (BH) 3DMRI via deformable image registration (DIR). Five volunteers participated in the study under an IRB-approved protocol. The 3D cine images with voxel size of 5×5×5 mm{sup 3} at two volumes per second (2Hz) were acquired coronally using a T1 fast field echo sequence, half-scan (0.8) acceleration, and SENSE (3) parallel imaging. Phase-encoding was set in the lateral direction to minimize motion artifacts. The BH image with voxel size of 2×2×2 mm{sup 3} was acquired using the same sequence within 10 seconds. A demons-based DIR program was employed to produce super-resolution 2Hz 4DMRI. Registration quality was visually assessed using difference images between TR 4DMRI and 3D cine and quantitatively assessed using average voxel correlation. The fidelity of the 3D cine images was assessed using a gel phantom and a 1D motion platform by comparing mobile and static images. Results: Owing to voxel intensity similarity using the same MRI scanning sequence, accurate DIR between FB and BH images is achieved. The voxel correlations between 3D cine and TR 4DMRI are greater than 0.92 in all cases and the difference images illustrate minimal residual error with little systematic patterns. The 3D cine images of the mobile gel phantom preserve object geometry with minimal scanning artifacts. Conclusion: The super-resolution time-resolved 4DMRI technique has been achieved via DIR, providing a potential solution for multi-breath motion assessment. Accurate DIR mapping has been achieved to map high-resolution BH images to low-resolution FB images, producing 2Hz volumetric high-resolution 4DMRI

  9. Fusion of lens-free microscopy and mobile-phone microscopy images for high-color-accuracy and high-resolution pathology imaging

    Science.gov (United States)

    Zhang, Yibo; Wu, Yichen; Zhang, Yun; Ozcan, Aydogan

    2017-03-01

    Digital pathology and telepathology require imaging tools with high-throughput, high-resolution and accurate color reproduction. Lens-free on-chip microscopy based on digital in-line holography is a promising technique towards these needs, as it offers a wide field of view (FOV >20 mm2) and high resolution with a compact, low-cost and portable setup. Color imaging has been previously demonstrated by combining reconstructed images at three discrete wavelengths in the red, green and blue parts of the visible spectrum, i.e., the RGB combination method. However, this RGB combination method is subject to color distortions. To improve the color performance of lens-free microscopy for pathology imaging, here we present a wavelet-based color fusion imaging framework, termed "digital color fusion microscopy" (DCFM), which digitally fuses together a grayscale lens-free microscope image taken at a single wavelength and a low-resolution and low-magnification color-calibrated image taken by a lens-based microscope, which can simply be a mobile phone based cost-effective microscope. We show that the imaging results of an H&E stained breast cancer tissue slide with the DCFM technique come very close to a color-calibrated microscope using a 40x objective lens with 0.75 NA. Quantitative comparison showed 2-fold reduction in the mean color distance using the DCFM method compared to the RGB combination method, while also preserving the high-resolution features of the lens-free microscope. Due to the cost-effective and field-portable nature of both lens-free and mobile-phone microscopy techniques, their combination through the DCFM framework could be useful for digital pathology and telepathology applications, in low-resource and point-of-care settings.

  10. 2D Unitary ESPRIT Based Super-Resolution Channel Estimation for Millimeter-Wave Massive MIMO with Hybrid Precoding

    KAUST Repository

    Liao, Anwen

    2017-11-01

    Millimeter-wave (mmWave) massive multiple-input multiple-output (MIMO) with hybrid precoding is a promising technique for the future 5G wireless communications. Due to a large number of antennas but a much smaller number of radio frequency (RF) chains, estimating the high-dimensional mmWave massive MIMO channel will bring the large pilot overhead. To overcome this challenge, this paper proposes a super-resolution channel estimation scheme based on two-dimensional (2D) unitary ESPRIT algorithm. By exploiting the angular sparsity of mmWave channels, the continuously distributed angle of arrivals/departures (AoAs/AoDs) can be jointly estimated with high accuracy. Specifically, by designing the uplink training signals at both base station (BS) and mobile station (MS), we first use low pilot overhead to estimate a low-dimensional effective channel, which has the same shift-invariance of array response as the high-dimensional mmWave MIMO channel to be estimated. From the low-dimensional effective channel, the superresolution estimates of AoAs and AoDs can be jointly obtained by exploiting the 2D unitary ESPRIT channel estimation algorithm. Furthermore, the associated path gains can be acquired based on the least squares (LS) criterion. Finally, we can reconstruct the high-dimensional mmWave MIMO channel according to the obtained AoAs, AoDs, and path gains. Simulation results have confirmed that the proposed scheme is superior to conventional schemes with a much lower pilot overhead.

  11. Improvement of Breast Cancer Detection Using Non-subsampled Contourlet Transform and Super-Resolution Technique in Mammographic Images

    Directory of Open Access Journals (Sweden)

    Fatemeh Pak

    2015-05-01

    Full Text Available Introduction Breast cancer is one of the most life-threatening conditions among women. Early detection of this disease is the only way to reduce the associated mortality rate. Mammography is a standard method for the early detection of breast cancer. Today, considering the importance of breast cancer detection, computer-aided detection techniques have been employed to increase the quality of mammographic images and help physicians reduce false positive rate (FPR. Materials and Methods In this study, a method was proposed for improving the quality of mammographic images to help radiologists establish a prompt and accurate diagnosis. The proposed approach included three major parts including pre-processing, feature extraction, and classification. In the pre-processing stage, the region of interest was determined and the image quality was improved by non-subsampled contourlet transform and super-resolution algorithm. In the feature extraction stage, some features of image components were extracted and skewness of each feature was calculated. Finally, a support vector machine was utilized to classify the features and determine the probability of benignity or malignancy of the disease. Results Based on the obtained results using Mammographic Image Analysis Society (MIAS database, the mean accuracy was estimated at 87.26% and maximum accuracy was 96.29%. Also, the mean and minimum FPRs were estimated at 9.55% and 2.87%, respectively.     Conclusion The results obtained using MIAS database indicated the superiority of the proposed method to other techniques. The reduced FPR in the proposed method was a significant finding in the present article.

  12. 2D Unitary ESPRIT Based Super-Resolution Channel Estimation for Millimeter-Wave Massive MIMO with Hybrid Precoding

    KAUST Repository

    Liao, Anwen; Gao, Zhen; Wu, Yongpeng; Wang, Hua; Alouini, Mohamed-Slim

    2017-01-01

    Millimeter-wave (mmWave) massive multiple-input multiple-output (MIMO) with hybrid precoding is a promising technique for the future 5G wireless communications. Due to a large number of antennas but a much smaller number of radio frequency (RF) chains, estimating the high-dimensional mmWave massive MIMO channel will bring the large pilot overhead. To overcome this challenge, this paper proposes a super-resolution channel estimation scheme based on two-dimensional (2D) unitary ESPRIT algorithm. By exploiting the angular sparsity of mmWave channels, the continuously distributed angle of arrivals/departures (AoAs/AoDs) can be jointly estimated with high accuracy. Specifically, by designing the uplink training signals at both base station (BS) and mobile station (MS), we first use low pilot overhead to estimate a low-dimensional effective channel, which has the same shift-invariance of array response as the high-dimensional mmWave MIMO channel to be estimated. From the low-dimensional effective channel, the superresolution estimates of AoAs and AoDs can be jointly obtained by exploiting the 2D unitary ESPRIT channel estimation algorithm. Furthermore, the associated path gains can be acquired based on the least squares (LS) criterion. Finally, we can reconstruct the high-dimensional mmWave MIMO channel according to the obtained AoAs, AoDs, and path gains. Simulation results have confirmed that the proposed scheme is superior to conventional schemes with a much lower pilot overhead.

  13. Inverse modeling applied to Scanning Capacitance Microscopy for improved spatial resolution and accuracy

    International Nuclear Information System (INIS)

    McMurray, J. S.; Williams, C. C.

    1998-01-01

    Scanning Capacitance Microscopy (SCM) is capable of providing two-dimensional information about dopant and carrier concentrations in semiconducting devices. This information can be used to calibrate models used in the simulation of these devices prior to manufacturing and to develop and optimize the manufacturing processes. To provide information for future generations of devices, ultra-high spatial accuracy (<10 nm) will be required. One method, which potentially provides a means to obtain these goals, is inverse modeling of SCM data. Current semiconducting devices have large dopant gradients. As a consequence, the capacitance probe signal represents an average over the local dopant gradient. Conversion of the SCM signal to dopant density has previously been accomplished with a physical model which assumes that no dopant gradient exists in the sampling area of the tip. The conversion of data using this model produces results for abrupt profiles which do not have adequate resolution and accuracy. A new inverse model and iterative method has been developed to obtain higher resolution and accuracy from the same SCM data. This model has been used to simulate the capacitance signal obtained from one and two-dimensional ideal abrupt profiles. This simulated data has been input to a new iterative conversion algorithm, which has recovered the original profiles in both one and two dimensions. In addition, it is found that the shape of the tip can significantly impact resolution. Currently SCM tips are found to degrade very rapidly. Initially the apex of the tip is approximately hemispherical, but quickly becomes flat. This flat region often has a radius of about the original hemispherical radius. This change in geometry causes the silicon directly under the disk to be sampled with approximately equal weight. In contrast, a hemispherical geometry samples most strongly the silicon centered under the SCM tip and falls off quickly with distance from the tip's apex. Simulation

  14. 3D imaging of hematoxylin and eosin stained thick tissues with a sub-femtoliter resolution by using Cr:forsterite-laser-based nonlinear microscopy (Conference Presentation)

    Science.gov (United States)

    Kao, Chien-Ting; Wei, Ming-Liang; Liao, Yi-Hua; Sun, Chi-Kuang

    2017-02-01

    Intraoperative assessment of excision tissues during cancer surgery is clinically important. The assessment is used to be guided by the examination for residual tumor with frozen pathology, while it is time consuming for preparation and is with low accuracy for diagnosis. Recently, reflection confocal microscopy (RCM) and nonlinear microscopy (NLM) were demonstrated to be promising methods for surgical border assessment. Intraoperative RCM imaging may enable detection of residual tumor directly on skin cancers patients during Mohs surgery. The assessment of benign and malignant breast pathologies in fresh surgical specimens was demonstrated by NLM. Without using hematoxylin and eosin (H and E) that are common dyes for histopathological diagnosis, RCM was proposed to image in vivo by using aluminum chloride for nuclear contrast on surgical wounds directly, while NLM was proposed to detect two photon fluorescence nuclear contrast from acrdine orange staining. In this paper, we propose and demonstrate 3D imaging of H and E stained thick tissues with a sub-femtoliter resolution by using Cr:forsterite-laser-based NLM. With a 1260 nm femtosecond Cr:forsterite laser as the excitation source, the hematoxylin will strongly enhance the third-harmonic generation (THG) signals, while eosin will illuminate strong fluorescence under three photon absorption. Compared with previous works, the 1260 nm excitation light provide high penetration and low photodamage to the exercised tissues so that the possibility to perform other follow-up examination will be preserved. The THG and three-photon process provides high nonlinearity so that the super resolution in 3D is now possible. The staining and the contrast of the imaging is also fully compatible with the current clinical standard on frozen pathology thus facilitate the rapid intraoperative assessment of excision tissues. This work is sponsored by National Health Research Institutes and supported by National Taiwan University

  15. Elemental mapping in achromatic atomic-resolution energy-filtered transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Forbes, B.D. [School of Physics, University of Melbourne, Parkville, VIC 3010 (Australia); Houben, L. [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons and Peter Gruenberg Institute, Forschungszentrum Jülich, D-52425 Jülich (Germany); Mayer, J. [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons and Peter Gruenberg Institute, Forschungszentrum Jülich, D-52425 Jülich (Germany); Central Facility for Electron Microscopy, RWTH Aachen University, D-52074 Aachen (Germany); Dunin-Borkowski, R.E. [Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons and Peter Gruenberg Institute, Forschungszentrum Jülich, D-52425 Jülich (Germany); Allen, L.J., E-mail: lja@unimelb.edu.au [School of Physics, University of Melbourne, Parkville, VIC 3010 (Australia)

    2014-12-15

    We present atomic-resolution energy-filtered transmission electron microscopy (EFTEM) images obtained with the chromatic-aberration-corrected FEI Titan PICO at the Ernst-Ruska Centre, Jülich, Germany. We find qualitative agreement between experiment and simulation for the background-subtracted EFTEM images of the Ti–L{sub 2,3} and O–K edges for a specimen of SrTiO{sub 3} oriented down the [110] zone axis. The simulations utilize the transition potential formulation for inelastic scattering, which permits a detailed investigation of contributions to the EFTEM image. We find that energy-filtered images of the Ti–L{sub 2,3} and O–K edges are lattice images and that the background-subtracted core-loss maps may not be directly interpretable as elemental maps. Simulations show that this is a result of preservation of elastic contrast, whereby the qualitative details of the image are determined primarily by elastic, coherent scattering. We show that this effect places a constraint on the range of specimen thicknesses which could theoretically yield directly useful elemental maps. In general, interpretation of EFTEM images is ideally accompanied by detailed simulations. - Highlights: • Achromatic atomic-resolution EFTEM images were obtained for STO 〈110〉. • Simulations were in qualitative agreement with Ti–L{sub 2,3} and O–K edge maps. • The experimental EFTEM maps are not directly interpretable as elemental maps. • Image intensities are strongly determined by preservation of elastic contrast. • Interpretation of EFTEM images is ideally accompanied by detailed simulations.

  16. High-resolution magnetic resonance imaging of rotator cuff tears using a microscopy coil. Noninvasive detection without intraarticular contrast material

    International Nuclear Information System (INIS)

    Hitachi, Shin; Takase, Kei; Higano, Shuichi; Takahashi, Shoki; Tanaka, Minoru; Tojo, Yuichi; Tabata, Shiro; Majima, Kazuhiro

    2011-01-01

    The aim of this study was to evaluate the feasibility of high-resolution magnetic resonance imaging (MRI) using a microscopy coil for the diagnosis of rotator cuff tears by comparing the method to conventional MRI and MRI arthrography. A total of 68 shoulders were prospectively studied using a 1.5-T MRI unit. Conventional MRI scans were obtained with a surface coil and high-resolution MRI scans with a microscopy coil. MRI arthrography was performed in 28 shoulders using a surface coil. MRI evaluation of tears of rotator cuff tendons was compared with arthroscopic findings and surgical results. The surgery revealed 40 full-thickness tears, 13 partial-thickness tears, and 15 intact cuffs. In all, 35 (88%) full-thickness tears were correctly diagnosed on conventional MRI and 40 (100%) on high-resolution MRI. MR arthrography delineated 11 of 12 (92%) full-thickness tears. Altogether, 5 (38%) of the partial-thickness tears were detected on conventional MRI, and 12 (92%) were clearly demonstrated on high-resolution MRI. MRI arthrography depicted three (60%) of five partial-thickness tears. High-resolution MRI showed higher sensitivity than conventional MRI (P<0.05) and had values equivalent to those of MRI arthrography for diagnosing partial-thickness tears. High-resolution MRI with a microscopy coil is a feasible, noninvasive technique for diagnosing rotator cuff tears. (author)

  17. Super-resolution inpainting

    Institute of Scientific and Technical Information of China (English)

    SHIH Timothy K; CHANG Rong-chi

    2005-01-01

    Image or video resources are often received in poor condition, mostly with noise or defects making the resources hard to read. We propose an effective algorithm based on digital image inpainting. The mechanism can be used in restoring images or video frames with very high noise or defect ratio (e.g., 90%). The algorithm is based on the concept of image subdivision and estimation of color variations. Noises inside blocks of different sizes are inpainted with different levels of surrounding information.The results showed that an almost unrecognizable image can be recovered with visually good result. The algorithm can be further extended for processing motion picture with high percentage of noise.

  18. Quantitative atomic resolution mapping using high-angle annular dark field scanning transmission electron microscopy

    International Nuclear Information System (INIS)

    Van Aert, S.; Verbeeck, J.; Erni, R.; Bals, S.; Luysberg, M.; Dyck, D. Van; Tendeloo, G. Van

    2009-01-01

    A model-based method is proposed to relatively quantify the chemical composition of atomic columns using high angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) images. The method is based on a quantification of the total intensity of the scattered electrons for the individual atomic columns using statistical parameter estimation theory. In order to apply this theory, a model is required describing the image contrast of the HAADF STEM images. Therefore, a simple, effective incoherent model has been assumed which takes the probe intensity profile into account. The scattered intensities can then be estimated by fitting this model to an experimental HAADF STEM image. These estimates are used as a performance measure to distinguish between different atomic column types and to identify the nature of unknown columns with good accuracy and precision using statistical hypothesis testing. The reliability of the method is supported by means of simulated HAADF STEM images as well as a combination of experimental images and electron energy-loss spectra. It is experimentally shown that statistically meaningful information on the composition of individual columns can be obtained even if the difference in averaged atomic number Z is only 3. Using this method, quantitative mapping at atomic resolution using HAADF STEM images only has become possible without the need of simultaneously recorded electron energy loss spectra.

  19. An extended model of electrons: experimental evidence from high-resolution scanning tunneling microscopy

    International Nuclear Information System (INIS)

    Hofer, Werner A

    2012-01-01

    In a recent paper we introduced a model of extended electrons, which is fully compatible with quantum mechanics in the formulation of Schrödinger. However, it contradicts the current interpretation of electrons as point-particles. Here, we show by a statistical analysis of high-resolution scanning tunneling microscopy (STM) experiments, that the interpretation of electrons as point particles and, consequently, the interpretation of the density of electron charge as a statistical quantity will lead to a conflict with the Heisenberg uncertainty principle. Given the precision in these experiments we find that the uncertainty principle would be violated by close to two orders of magnitude, if this interpretation were correct. We are thus forced to conclude that the density of electron charge is a physically real, i.e. in principle precisely measurable quantity, as derived in a recent paper. Experimental evidence to the contrary, in particular high-energy scattering experiments, is briefly discussed. The finding is expected to have wide implications in condensed matter physics, chemistry, and biology, scientific disciplines which are based on the properties and interactions of electrons.

  20. EnLightenment: High resolution smartphone microscopy as an educational and public engagement platform

    Science.gov (United States)

    Wicks, Laura C.; Cairns, Gemma S.; Melnyk, Jacob; Bryce, Scott; Duncan, Rory R.; Dalgarno, Paul A.

    2018-01-01

    We developed a simple, cost-effective smartphone microscopy platform for use in educational and public engagement programs. We demonstrated its effectiveness, and potential for citizen science through a national imaging initiative, EnLightenment. The cost effectiveness of the instrument allowed for the program to deliver over 500 microscopes to more than 100 secondary schools throughout Scotland, targeting 1000’s of 12-14 year olds. Through careful, quantified, selection of a high power, low-cost objective lens, our smartphone microscope has an imaging resolution of microns, with a working distance of 3 mm. It is therefore capable of imaging single cells and sub-cellular features, and retains usability for young children. The microscopes were designed in kit form and provided an interdisciplinary educational tool. By providing full lesson plans and support material, we developed a framework to explore optical design, microscope performance, engineering challenges on construction and real-world applications in life sciences, biological imaging, marine biology, art, and technology. A national online imaging competition framed EnLightenment ; with over 500 high quality images submitted of diverse content, spanning multiple disciplines. With examples of cellular and sub-cellular features clearly identifiable in some submissions, we show how young public can use these instruments for research-level imaging applications, and the potential of the instrument for citizen science programs. PMID:29623296

  1. An improved image alignment procedure for high-resolution transmission electron microscopy.

    Science.gov (United States)

    Lin, Fang; Liu, Yan; Zhong, Xiaoyan; Chen, Jianghua

    2010-06-01

    Image alignment is essential for image processing methods such as through-focus exit-wavefunction reconstruction and image averaging in high-resolution transmission electron microscopy. Relative image displacements exist in any experimentally recorded image series due to the specimen drifts and image shifts, hence image alignment for correcting the image displacements has to be done prior to any further image processing. The image displacement between two successive images is determined by the correlation function of the two relatively shifted images. Here it is shown that more accurate image alignment can be achieved by using an appropriate aperture to filter the high-frequency components of the images being aligned, especially for a crystalline specimen with little non-periodic information. For the image series of crystalline specimens with little amorphous, the radius of the filter aperture should be as small as possible, so long as it covers the innermost lattice reflections. Testing with an experimental through-focus series of Si[110] images, the accuracies of image alignment with different correlation functions are compared with respect to the error functions in through-focus exit-wavefunction reconstruction based on the maximum-likelihood method. Testing with image averaging over noisy experimental images from graphene and carbon-nanotube samples, clear and sharp crystal lattice fringes are recovered after applying optimal image alignment. Copyright 2010 Elsevier Ltd. All rights reserved.

  2. Method for local temperature measurement in a nanoreactor for in situ high-resolution electron microscopy.

    Science.gov (United States)

    Vendelbo, S B; Kooyman, P J; Creemer, J F; Morana, B; Mele, L; Dona, P; Nelissen, B J; Helveg, S

    2013-10-01

    In situ high-resolution transmission electron microscopy (TEM) of solids under reactive gas conditions can be facilitated by microelectromechanical system devices called nanoreactors. These nanoreactors are windowed cells containing nanoliter volumes of gas at ambient pressures and elevated temperatures. However, due to the high spatial confinement of the reaction environment, traditional methods for measuring process parameters, such as the local temperature, are difficult to apply. To address this issue, we devise an electron energy loss spectroscopy (EELS) method that probes the local temperature of the reaction volume under inspection by the electron beam. The local gas density, as measured using quantitative EELS, is combined with the inherent relation between gas density and temperature, as described by the ideal gas law, to obtain the local temperature. Using this method we determined the temperature gradient in a nanoreactor in situ, while the average, global temperature was monitored by a traditional measurement of the electrical resistivity of the heater. The local gas temperatures had a maximum of 56 °C deviation from the global heater values under the applied conditions. The local temperatures, obtained with the proposed method, are in good agreement with predictions from an analytical model. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. High resolution electron microscopy of the triply incommensurate phase of 2H-TaSe2

    Science.gov (United States)

    Onozuka, Takashi; Otsuka, Nobuo; Sato, Hiroshi

    1986-09-01

    The triply incommensurate phase of 2H-TaSe2 obtained by cooling from the normal phase was investigated by transmission electron microscopy between 87 and 113 K with the resolution of 3 Å, one order of magnitude better than earlier experiments. Moirélike patterns observed in this phase were confirmed to be interference fringes due to the first- and second-order diffraction beams (with small separation and possibly with higher-order diffraction beams) from the incommensurate structure and were not due to the dark-field diffraction contrast of domains of the commensurate structure as interpreted earlier. Lattice fringes (~9 Å) of this modulated phase do not show any discontinuity across the boundaries of regions of different contrasts of the moirélike fringes which is expected from domain boundaries. Instead, a periodic change in the spacing of the lattice fringes (phase-slip region) expected from the superposition of split superlattice spots in forming the lattice image is observed. This is what is believed to be the first direct observation of the existence of the phase-slip region which is also expected from the discommensuration theory. A series of observations presented here thus shows that the triply incommensurate phase is intrinsically incommensurate and suggests the need for a modification of interpretations of this phase in terms of the double honeycomb discommensuration model.

  4. Atom-counting in High Resolution Electron Microscopy:TEM or STEM - That's the question.

    Science.gov (United States)

    Gonnissen, J; De Backer, A; den Dekker, A J; Sijbers, J; Van Aert, S

    2017-03-01

    In this work, a recently developed quantitative approach based on the principles of detection theory is used in order to determine the possibilities and limitations of High Resolution Scanning Transmission Electron Microscopy (HR STEM) and HR TEM for atom-counting. So far, HR STEM has been shown to be an appropriate imaging mode to count the number of atoms in a projected atomic column. Recently, it has been demonstrated that HR TEM, when using negative spherical aberration imaging, is suitable for atom-counting as well. The capabilities of both imaging techniques are investigated and compared using the probability of error as a criterion. It is shown that for the same incoming electron dose, HR STEM outperforms HR TEM under common practice standards, i.e. when the decision is based on the probability function of the peak intensities in HR TEM and of the scattering cross-sections in HR STEM. If the atom-counting decision is based on the joint probability function of the image pixel values, the dependence of all image pixel intensities as a function of thickness should be known accurately. Under this assumption, the probability of error may decrease significantly for atom-counting in HR TEM and may, in theory, become lower as compared to HR STEM under the predicted optimal experimental settings. However, the commonly used standard for atom-counting in HR STEM leads to a high performance and has been shown to work in practice. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Dynamic electrostatic force microscopy technique for the study of electrical properties with improved spatial resolution

    International Nuclear Information System (INIS)

    Maragliano, C; Heskes, D; Stefancich, M; Chiesa, M; Souier, T

    2013-01-01

    The need to resolve the electrical properties of confined structures (CNTs, quantum dots, nanorods, etc) is becoming increasingly important in the field of electronic and optoelectronic devices. Here we propose an approach based on amplitude modulated electrostatic force microscopy to obtain measurements at small tip–sample distances, where highly nonlinear forces are present. We discuss how this improves the lateral resolution of the technique and allows probing of the electrical and surface properties. The complete force field at different tip biases is employed to derive the local work function difference. Then, by appropriately biasing the tip–sample system, short-range forces are reconstructed. The short-range component is then separated from the generic tip–sample force in order to recover the pure electrostatic contribution. This data can be employed to derive the tip–sample capacitance curve and the sample dielectric constant. After presenting a theoretical model that justifies the need for probing the electrical properties of the sample in the vicinity of the surface, the methodology is presented in detail and verified experimentally. (paper)

  6. Micaceous occlusions in kaolinite observed by ultramicrotomy and high resolution electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lee, S Y [Univ. of Wisconsin, Madison; Jackson, M L; Brown, J L

    1975-01-01

    The layer structure of kaolinite from Twiggs, Georgia, and fire-clay type kaolinite (Frantex B, from France, particle size separates 2 0.2 ..mu..m was studied by high resolution electron microscopy after embedment in Spurr low-viscosity Epoxy media and thin sectioning normal to the (001) planes by an ultramicrotome. Images of the (001) planes (viewed edge-on) of both kaolinites were spaced at 7 A and generally aligned in parallel, with occasional bending into more widely spaced images of about 10 A interval. Some of the 10 A images converged to 7 A at one or both ends, forming ellipse-shaped islands 80 to 130 A thick and 300 to 500 A long. The island areas and interleaved 10 A layers between 7 A layers may represent a residue of incomplete weathering of mica to kaolinite. The proportions of micaceous occlusions were too small and the layer sequences too irregular to be detected by X-ray diffraction. The lateral continuity of the layers through the 7-10-7 A sequence in a kaolinite particle would partially interrupt or prevent expansion in dimethyl sulfoxide (DMSO) and other kaolinite intercalating media. Discrete mica particles were also observed with parallel images at 10 A, as impurities in both kaolinites. The small K content of the chemical analyses of the kaolinite samples is accounted for as interlayer K, not only in discrete mica particles but also in the micaceous occlusions.

  7. Nanometer-resolution electron microscopy through micrometers-thick water layers

    Energy Technology Data Exchange (ETDEWEB)

    Jonge, Niels de, E-mail: niels.de.jonge@vanderbilt.edu [Vanderbilt University Medical Center, Department of Molecular Physiology and Biophysics, Nashville, TN 37232-0615 (United States); Oak Ridge National Laboratory, Materials Science and Technology Division, Oak Ridge, TN 37831-6064 (United States); Poirier-Demers, Nicolas; Demers, Hendrix [Universite de Sherbrooke, Electrical and Computer Engineering, Sherbrooke, Quebec J1K 2R1 (Canada); Peckys, Diana B. [Oak Ridge National Laboratory, Materials Science and Technology Division, Oak Ridge, TN 37831-6064 (United States); University of Tennessee, Center for Environmental Biotechnology, Knoxville, TN 37996-1605 (United States); Drouin, Dominique [Universite de Sherbrooke, Electrical and Computer Engineering, Sherbrooke, Quebec J1K 2R1 (Canada)

    2010-08-15

    Scanning transmission electron microscopy (STEM) was used to image gold nanoparticles on top of and below saline water layers of several micrometers thickness. The smallest gold nanoparticles studied had diameters of 1.4 nm and were visible for a liquid thickness of up to 3.3 {mu}m. The imaging of gold nanoparticles below several micrometers of liquid was limited by broadening of the electron probe caused by scattering of the electron beam in the liquid. The experimental data corresponded to analytical models of the resolution and of the electron probe broadening as function of the liquid thickness. The results were also compared with Monte Carlo simulations of the STEM imaging on modeled specimens of similar geometry and composition as used for the experiments. Applications of STEM imaging in liquid can be found in cell biology, e.g., to study tagged proteins in whole eukaryotic cells in liquid and in materials science to study the interaction of solid:liquid interfaces at the nanoscale.

  8. Enhancing resolution and contrast in second-harmonic generation microscopy using an advanced maximum likelihood estimation restoration method

    Science.gov (United States)

    Sivaguru, Mayandi; Kabir, Mohammad M.; Gartia, Manas Ranjan; Biggs, David S. C.; Sivaguru, Barghav S.; Sivaguru, Vignesh A.; Berent, Zachary T.; Wagoner Johnson, Amy J.; Fried, Glenn A.; Liu, Gang Logan; Sadayappan, Sakthivel; Toussaint, Kimani C.

    2017-02-01

    Second-harmonic generation (SHG) microscopy is a label-free imaging technique to study collagenous materials in extracellular matrix environment with high resolution and contrast. However, like many other microscopy techniques, the actual spatial resolution achievable by SHG microscopy is reduced by out-of-focus blur and optical aberrations that degrade particularly the amplitude of the detectable higher spatial frequencies. Being a two-photon scattering process, it is challenging to define a point spread function (PSF) for the SHG imaging modality. As a result, in comparison with other two-photon imaging systems like two-photon fluorescence, it is difficult to apply any PSF-engineering techniques to enhance the experimental spatial resolution closer to the diffraction limit. Here, we present a method to improve the spatial resolution in SHG microscopy using an advanced maximum likelihood estimation (AdvMLE) algorithm to recover the otherwise degraded higher spatial frequencies in an SHG image. Through adaptation and iteration, the AdvMLE algorithm calculates an improved PSF for an SHG image and enhances the spatial resolution by decreasing the full-width-at-halfmaximum (FWHM) by 20%. Similar results are consistently observed for biological tissues with varying SHG sources, such as gold nanoparticles and collagen in porcine feet tendons. By obtaining an experimental transverse spatial resolution of 400 nm, we show that the AdvMLE algorithm brings the practical spatial resolution closer to the theoretical diffraction limit. Our approach is suitable for adaptation in micro-nano CT and MRI imaging, which has the potential to impact diagnosis and treatment of human diseases.

  9. High-resolution MR imaging of triangular fibrocartilage complex (TFCC): comparison of microscopy coils and a conventional small surface coil

    Energy Technology Data Exchange (ETDEWEB)

    Yoshioka, Hiroshi [Department of Radiology, University of Tsukuba, Tsukuba (Japan); Department of Radiology, Brigham and Women' s Hospital, 75 Francis Street, 02115, Boston, MA (United States); Ueno, Teruko; Itai, Yuji [Department of Radiology, University of Tsukuba, Tsukuba (Japan); Tanaka, Toshikazu [Department of Orthopedic Surgery, Tsukuba Kinen Hospital, Tsukuba (Japan); Shindo, Masashi [Tsukuba University Hospital, Tsukuba (Japan)

    2003-10-01

    To compare MR images of the triangular fibrocartilage complex (TFCC) using microscopy coils with those using a conventional surface coil qualitatively and quantitatively. Proton density-weighted images and T2*-weighted images of the TFCC from ten normal volunteers were obtained with a conventional surface coil (C4 coil; 80 mm in diameter), a 47-mm microscopy surface coil and a 23-mm microscopy surface coil at 1.5 T. Qualitative image analysis of MR images with three coils was performed by two radiologists who assigned one of five numerical scores (0, nonvisualization; 1, poor; 2, average; 3, good; 4, excellent) for five TFCC components, which were disc proper, triangular ligament, meniscus homologue, ulnotriquetral and ulnolunate ligament. Quantitative analysis included the signal-to-noise ratio (S/N) of the disc proper of TFCC, the lunate cartilage, the lunate bone and the contrast-noise-ratio (C/N) between articular cartilage and disc proper or bone marrow were measured. All structures show higher scores qualitatively on MR with microscopy coils than those with a C4 coil, and the difference was significant with the exception of the ulnolunate ligament. MR with microscopy coils showed significantly higher S/N values than those with a conventional surface coil (P<0.05 to P<0.001). T2*-weighted images using microscopy coils showed significantly higher cartilage-disc proper C/N and cartilage-bone marrow C/N (P<0.01 to P<0.001). On proton density-weighted images, the C/N between cartilage and disc proper with two microscopy coils was significantly higher (P<0.01) than that with a conventional coil. High-resolution MR images of the normal wrist using microscopy coils were superior to those using a conventional surface coil qualitatively and quantitatively. High-resolution MR imaging with a microscopy coil would be a promising method to diagnose TFCC lesions. (orig.)

  10. Stimulated-emission pumping enabling sub-diffraction-limited spatial resolution in coherent anti-Stokes Raman scattering microscopy

    NARCIS (Netherlands)

    Cleff, C.; Gross, P.; Fallnich, C.; Offerhaus, Herman L.; Herek, Jennifer Lynn; Kruse, K.; Beeker, W.P.; Lee, Christopher James; Boller, Klaus J.

    2013-01-01

    We present a theoretical investigation of stimulated emission pumping to achieve sub-diffraction-limited spatial resolution in coherent anti-Stokes Raman scattering (CARS) microscopy. A pair of control light fields is used to prepopulate the Raman state involved in the CARS process prior to the CARS

  11. Length-extension resonator as a force sensor for high-resolution frequency-modulation atomic force microscopy in air.

    Science.gov (United States)

    Beyer, Hannes; Wagner, Tino; Stemmer, Andreas

    2016-01-01

    Frequency-modulation atomic force microscopy has turned into a well-established method to obtain atomic resolution on flat surfaces, but is often limited to ultra-high vacuum conditions and cryogenic temperatures. Measurements under ambient conditions are influenced by variations of the dew point and thin water layers present on practically every surface, complicating stable imaging with high resolution. We demonstrate high-resolution imaging in air using a length-extension resonator operating at small amplitudes. An additional slow feedback compensates for changes in the free resonance frequency, allowing stable imaging over a long period of time with changing environmental conditions.

  12. Single-shot full resolution region-of-interest (ROI) reconstruction in image plane digital holographic microscopy

    Science.gov (United States)

    Singh, Mandeep; Khare, Kedar

    2018-05-01

    We describe a numerical processing technique that allows single-shot region-of-interest (ROI) reconstruction in image plane digital holographic microscopy with full pixel resolution. The ROI reconstruction is modelled as an optimization problem where the cost function to be minimized consists of an L2-norm squared data fitting term and a modified Huber penalty term that are minimized alternately in an adaptive fashion. The technique can provide full pixel resolution complex-valued images of the selected ROI which is not possible to achieve with the commonly used Fourier transform method. The technique can facilitate holographic reconstruction of individual cells of interest from a large field-of-view digital holographic microscopy data. The complementary phase information in addition to the usual absorption information already available in the form of bright field microscopy can make the methodology attractive to the biomedical user community.

  13. Linear versus non-linear structural information limit in high-resolution transmission electron microscopy

    International Nuclear Information System (INIS)

    Van Aert, S.; Chen, J.H.; Van Dyck, D.

    2010-01-01

    A widely used performance criterion in high-resolution transmission electron microscopy (HRTEM) is the information limit. It corresponds to the inverse of the maximum spatial object frequency that is linearly transmitted with sufficient intensity from the exit plane of the object to the image plane and is limited due to partial temporal coherence. In practice, the information limit is often measured from a diffractogram or from Young's fringes assuming a weak phase object scattering beyond the inverse of the information limit. However, for an aberration corrected electron microscope, with an information limit in the sub-angstrom range, weak phase objects are no longer applicable since they do not scatter sufficiently in this range. Therefore, one relies on more strongly scattering objects such as crystals of heavy atoms observed along a low index zone axis. In that case, dynamical scattering becomes important such that the non-linear and linear interaction may be equally important. The non-linear interaction may then set the experimental cut-off frequency observed in a diffractogram. The goal of this paper is to quantify both the linear and the non-linear information transfer in terms of closed form analytical expressions. Whereas the cut-off frequency set by the linear transfer can be directly related with the attainable resolution, information from the non-linear transfer can only be extracted using quantitative, model-based methods. In contrast to the historic definition of the information limit depending on microscope parameters only, the expressions derived in this paper explicitly incorporate their dependence on the structure parameters as well. In order to emphasize this dependence and to distinguish from the usual information limit, the expressions derived for the inverse cut-off frequencies will be referred to as the linear and non-linear structural information limit. The present findings confirm the well-known result that partial temporal coherence has

  14. Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy

    DEFF Research Database (Denmark)

    Clausen, M. P.; Colin-York, H.; Schneider, Falk

    2017-01-01

    and accurately measure the density distribution of the cortical actin cytoskeleton and the distance between the actin cortex and the membrane in live Jurkat T-cells. We found an asymmetric cortical actin density distribution with a mean width of 230 (+105/-125) nm. The spatial distances measured between...

  15. Optical Saturation as a Versatile Tool to Enhance Resolution in Confocal Microscopy

    Czech Academy of Sciences Publication Activity Database

    Humpolíčková, Jana; Benda, Aleš; Enderlein, J.

    2009-01-01

    Roč. 97, č. 9 (2009), s. 2623-2629 ISSN 0006-3495 R&D Projects: GA AV ČR KJB400400904; GA MŠk(CZ) LC06063 Institutional research plan: CEZ:AV0Z40400503 Keywords : fluorescence microscopy * reconstruction microscopy * cassettes Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 4.390, year: 2009

  16. Phase contrast scanning transmission electron microscopy imaging of light and heavy atoms at the limit of contrast and resolution.

    Science.gov (United States)

    Yücelen, Emrah; Lazić, Ivan; Bosch, Eric G T

    2018-02-08

    Using state of the art scanning transmission electron microscopy (STEM) it is nowadays possible to directly image single atomic columns at sub-Å resolution. In standard (high angle) annular dark field STEM ((HA)ADF-STEM), however, light elements are usually invisible when imaged together with heavier elements in one image. Here we demonstrate the capability of the recently introduced Integrated Differential Phase Contrast STEM (iDPC-STEM) technique to image both light and heavy atoms in a thin sample at sub-Å resolution. We use the technique to resolve both the Gallium and Nitrogen dumbbells in a GaN crystal in [[Formula: see text

  17. Analysis and Calibration of in situ scanning tunnelling microscopy Images with atomic Resolution Influenced by Surface Drift Phenomena

    DEFF Research Database (Denmark)

    Andersen, Jens Enevold Thaulov; Møller, Per

    1994-01-01

    The influence of surface drift velocities on in situ scanning tunnelling microscopy (STM) experiments with atomic resolution is analysed experimentally and mathematically. Constant drift velocities much smaller than the speed of scanning can in many in situ STM experiments with atomic resolution ...... as well as the vectors of the non-distorted surface lattice can be determined. The calibration of distances can thus be carried out also when the image is influenced by drift. Results with gold surfaces and graphite surfaces are analysed and discussed....

  18. Concepts in Light Microscopy of Viruses

    Science.gov (United States)

    Witte, Robert; Georgi, Fanny

    2018-01-01

    Viruses threaten humans, livestock, and plants, and are difficult to combat. Imaging of viruses by light microscopy is key to uncover the nature of known and emerging viruses in the quest for finding new ways to treat viral disease and deepening the understanding of virus–host interactions. Here, we provide an overview of recent technology for imaging cells and viruses by light microscopy, in particular fluorescence microscopy in static and live-cell modes. The review lays out guidelines for how novel fluorescent chemical probes and proteins can be used in light microscopy to illuminate cells, and how they can be used to study virus infections. We discuss advantages and opportunities of confocal and multi-photon microscopy, selective plane illumination microscopy, and super-resolution microscopy. We emphasize the prevalent concepts in image processing and data analyses, and provide an outlook into label-free digital holographic microscopy for virus research. PMID:29670029

  19. High resolution transmission electron microscopy studies of {sigma} phase in Ni-based single crystal superalloys

    Energy Technology Data Exchange (ETDEWEB)

    Sun Fei [Key Laboratory of Liquid Structure and Heredity of Materials, Ministry of Education, Shandong University, Jinan 250061 (China); Zhang Jianxin, E-mail: jianxin@sdu.edu.cn [Key Laboratory of Liquid Structure and Heredity of Materials, Ministry of Education, Shandong University, Jinan 250061 (China); Liu Pan [Institute of Microstructure and Property of Advanced Materials, Beijing University of Technology, Beijing 100124 (China); Feng Qiang [National Center for Materials Service Safety, University of Science and Technology Beijing, Beijing 100083 (China); State Key Laboratory for Advanced Metals and Materials, University of Science and Technology Beijing, Beijing 100083 (China); Han Xiaodong; Mao Shengcheng [Institute of Microstructure and Property of Advanced Materials, Beijing University of Technology, Beijing 100124 (China)

    2012-09-25

    Graphical abstract: (a) TEM micrograph of {sigma} phase; (b) HRTEM image of {sigma}/{gamma} interface corresponding to the area of the white frame in (a); (c) an enlarged image of area from the white frame in (b). The combination of {sigma}/{gamma} interface appears very well, and a two-atomic-layer step is shown on the {sigma}/{gamma} interface. In addition, {sigma} phase has the orientation relationship of [0 0 1]{sub {gamma}}//[1 1 2{sup Macron }]{sub {sigma}}, (2{sup Macron} 2 0){sub {gamma}}//(1{sup Macron} 1 0){sub {sigma}}, (2{sup Macron }2{sup Macron} 0){sub {gamma}}//(1 1 1){sub {sigma}}; [0 1 1]{sub {gamma}}//[1 1 0]{sub {sigma}}, (1 1{sup Macron} 1){sub {gamma}}//(0 0 1{sup Macron }){sub {sigma}} with the {gamma} phase. Highlights: Black-Right-Pointing-Pointer Elemental characteristic of {sigma} phase is studied by HAADF techniques and EDS analysis. Black-Right-Pointing-Pointer Interfacial characteristics of {sigma}/{gamma} interface are revealed by HRTEM. Black-Right-Pointing-Pointer An atomic structural {sigma}/{gamma} interface with a two-atomic-layer step has been proposed. - Abstract: By means of high resolution transmission electron microscopy (HRTEM) and high-angle annular dark-field image technique (HAADF), morphological of plate-shaped {sigma} phase and interfacial characteristics between plate-shaped {sigma} phase and {gamma} phase in Ni-based single crystal superalloys have been studied. On the basis of HRTEM observations, an atomic structural interface between {sigma} phase and {gamma} phase with a step has been proposed. {sigma} Phase has the relationship of [0 0 1]{sub {gamma}}//[1 1 2{sup Macron }]{sub {sigma}}, (2{sup Macron} 2 0){sub {gamma}}//(1{sup Macron} 1 0){sub {sigma},} (2{sup Macron }2{sup Macron} 0){sub {gamma}}//(1 1 1){sub {sigma}}; [0 1 1]{sub {gamma}}//[1 1 0]{sub {sigma}}, (1 1{sup Macron} 1){sub {gamma}}//(0 0 1{sup Macron }){sub {sigma}} with the {gamma} phase. The compositional characteristics of the {sigma} phase which

  20. Fluorescence photooxidation with eosin: a method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy

    Science.gov (United States)

    1994-01-01

    A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate. PMID:7519623

  1. 3D imaging of cement-based materials at submicron resolution by combining laser scanning confocal microscopy with serial sectioning.

    Science.gov (United States)

    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. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  2. High resolution electron microscopy studies of interfaces between Al2O3 substrates and MBE grown Nb films

    International Nuclear Information System (INIS)

    Mayer, J.; Ruhle, M.; Dura, J.; Flynn, C.P.

    1991-01-01

    This paper reports on single crystal niobium films grown by Molecular Beam Epitaxy (MBE) on (001) S sapphire substrates. Cross-sectional specimens with thickness of 2 O 3 interface could be investigated by high resolution electron microscopy (HREM). The orientation relationship between the metal film and the ceramic substrate was verified by selected area diffraction: (111) Nb parallel (0001) S and [1 bar 10] Nb parallel [2 bar 1 bar 10] S . The atomistic structure of the interface was identified by HREM

  3. Torsional resonance mode magnetic force microscopy: enabling higher lateral resolution magnetic imaging without topography-related effects

    International Nuclear Information System (INIS)

    Kaidatzis, A; García-Martín, J M

    2013-01-01

    We present experimental work that reveals the benefits of performing magnetic force microscopy measurements employing the torsional resonance mode of cantilever oscillation. This approach provides two clear advantages: the ability of performing magnetic imaging without topography-related interference and the significant lateral resolution improvement (approximately 15%). We believe that this work demonstrates a significant improvement to a versatile magnetic imaging technique widely used in academia and in industry. (paper)

  4. Stimulated emission depletion microscopy resolves individual nitrogen vacancy centers in diamond nanocrystals.

    OpenAIRE

    Arroyo Camejo, S.; Adam, M.; Besbes, M.; Hugonin, J.; Jaques, V.; Greffet, J.; Roch, J.; Hell, S.; Treussart, F.

    2013-01-01

    Nitrogen-vacancy (NV) color centers in nanodiamonds are highly promising for bioimaging and sensing. However, resolving individual NV centers within nanodiamond particles and the controlled addressing and readout of their spin state has remained a major challenge. Spatially stochastic super-resolution techniques cannot provide this capability in principle, whereas coordinate-controlled super-resolution imaging methods, like stimulated emission depletion (STED) microscopy, have been predicted ...

  5. Automatic Segmentation of Fluorescence Lifetime Microscopy Images of Cells Using Multi-Resolution Community Detection -A First Study

    Science.gov (United States)

    Hu, Dandan; Sarder, Pinaki; Ronhovde, Peter; Orthaus, Sandra; Achilefu, Samuel; Nussinov, Zohar

    2014-01-01

    Inspired by a multi-resolution community detection (MCD) based network segmentation method, we suggest an automatic method for segmenting fluorescence lifetime (FLT) imaging microscopy (FLIM) images of cells in a first pilot investigation on two selected images. The image processing problem is framed as identifying segments with respective average FLTs against the background in FLIM images. The proposed method segments a FLIM image for a given resolution of the network defined using image pixels as the nodes and similarity between the FLTs of the pixels as the edges. In the resulting segmentation, low network resolution leads to larger segments, and high network resolution leads to smaller segments. Further, using the proposed method, the mean-square error (MSE) in estimating the FLT segments in a FLIM image was found to consistently decrease with increasing resolution of the corresponding network. The MCD method appeared to perform better than a popular spectral clustering based method in performing FLIM image segmentation. At high resolution, the spectral segmentation method introduced noisy segments in its output, and it was unable to achieve a consistent decrease in MSE with increasing resolution. PMID:24251410

  6. Optimizing low-light microscopy with back-illuminated electron multiplying charge-coupled device: enhanced sensitivity, speed, and resolution.

    Science.gov (United States)

    Coates, Colin G; Denvir, Donal J; McHale, Noel G; Thornbury, Keith D; Hollywood, Mark A

    2004-01-01

    The back-illuminated electron multiplying charge-coupled device (EMCCD) camera is having a profound influence on the field of low-light dynamic cellular microscopy, combining highest possible photon collection efficiency with the ability to virtually eliminate the readout noise detection limit. We report here the use of this camera, in 512 x 512 frame-transfer chip format at 10-MHz pixel readout speed, in optimizing a demanding ultra-low-light intracellular calcium flux microscopy setup. The arrangement employed includes a spinning confocal Nipkow disk, which, while facilitating the need to both generate images at very rapid frame rates and minimize background photons, yields very weak signals. The challenge for the camera lies not just in detecting as many of these scarce photons as possible, but also in operating at a frame rate that meets the temporal resolution requirements of many low-light microscopy approaches, a particular demand of smooth muscle calcium flux microscopy. Results presented illustrate both the significant sensitivity improvement offered by this technology over the previous standard in ultra-low-light CCD detection, the GenIII+intensified charge-coupled device (ICCD), and also portray the advanced temporal and spatial resolution capabilities of the EMCCD. Copyright 2004 Society of Photo-Optical Instrumentation Engineers.

  7. Light-Induced Fluorescence Modulation of Quantum Dot-Crystal Violet Conjugates: Stochastic Off-On-Off Cycles for Multicolor Patterning and Super-Resolution.

    Science.gov (United States)

    Jung, Sungwook; Park, Joonhyuck; Bang, Jiwon; Kim, Jae-Yeol; Kim, Cheolhee; Jeon, Yongmoon; Lee, Seung Hwan; Jin, Ho; Choi, Sukyung; Kim, Bomi; Lee, Woo Jin; Pack, Chan-Gi; Lee, Jong-Bong; Lee, Nam Ki; Kim, Sungjee

    2017-06-07

    Photoswitching or modulation of quantum dots (QDs) can be promising for many fields that include display, memory, and super-resolution imaging. However, such modulations have mostly relied on photomodulations of conjugated molecules in QD vicinity, which typically require high power of high energy photons at UV. We report a visible light-induced facile modulation route for QD-dye conjugates. QD crystal violets conjugates (QD-CVs) were prepared and the crystal violet (CV) molecules on QD quenched the fluorescence efficiently. The fluorescence of QD-CVs showed a single cycle of emission burst as they go through three stages of (i) initially quenched "off" to (ii) photoactivated "on" as the result of chemical change of CVs induced by photoelectrons from QD and (iii) back to photodarkened "off" by radical-associated reactions. Multicolor on-demand photopatterning was demonstrated using QD-CV solid films. QD-CVs were introduced into cells, and excitation with visible light yielded photomodulation from "off" to "on" and "off" by nearly ten fold. Individual photoluminescence dynamics of QD-CVs was investigated using fluorescence correlation spectroscopy and single QD emission analysis, which revealed temporally stochastic photoactivations and photodarkenings. Exploiting the stochastic fluorescence burst of QD-CVs, simultaneous multicolor super-resolution localizations were demonstrated.

  8. Interactive local super-resolution reconstruction of whole-body MRI mouse data: a pilot study with applications to bone and kidney metastases.

    Directory of Open Access Journals (Sweden)

    Oleh Dzyubachyk

    Full Text Available In small animal imaging studies, when the locations of the micro-structures of interest are unknown a priori, there is a simultaneous need for full-body coverage and high resolution. In MRI, additional requirements to image contrast and acquisition time will often make it impossible to acquire such images directly. Recently, a resolution enhancing post-processing technique called super-resolution reconstruction (SRR has been demonstrated to improve visualization and localization of micro-structures in small animal MRI by combining multiple low-resolution acquisitions. However, when the field-of-view is large relative to the desired voxel size, solving the SRR problem becomes very expensive, in terms of both memory requirements and computation time. In this paper we introduce a novel local approach to SRR that aims to overcome the computational problems and allow researchers to efficiently explore both global and local characteristics in whole-body small animal MRI. The method integrates state-of-the-art image processing techniques from the areas of articulated atlas-based segmentation, planar reformation, and SRR. A proof-of-concept is provided with two case studies involving CT, BLI, and MRI data of bone and kidney tumors in a mouse model. We show that local SRR-MRI is a computationally efficient complementary imaging modality for the precise characterization of tumor metastases, and that the method provides a feasible high-resolution alternative to conventional MRI.

  9. Interactive local super-resolution reconstruction of whole-body MRI mouse data: a pilot study with applications to bone and kidney metastases.

    Science.gov (United States)

    Dzyubachyk, Oleh; Khmelinskii, Artem; Plenge, Esben; Kok, Peter; Snoeks, Thomas J A; Poot, Dirk H J; Löwik, Clemens W G M; Botha, Charl P; Niessen, Wiro J; van der Weerd, Louise; Meijering, Erik; Lelieveldt, Boudewijn P F

    2014-01-01

    In small animal imaging studies, when the locations of the micro-structures of interest are unknown a priori, there is a simultaneous need for full-body coverage and high resolution. In MRI, additional requirements to image contrast and acquisition time will often make it impossible to acquire such images directly. Recently, a resolution enhancing post-processing technique called super-resolution reconstruction (SRR) has been demonstrated to improve visualization and localization of micro-structures in small animal MRI by combining multiple low-resolution acquisitions. However, when the field-of-view is large relative to the desired voxel size, solving the SRR problem becomes very expensive, in terms of both memory requirements and computation time. In this paper we introduce a novel local approach to SRR that aims to overcome the computational problems and allow researchers to efficiently explore both global and local characteristics in whole-body small animal MRI. The method integrates state-of-the-art image processing techniques from the areas of articulated atlas-based segmentation, planar reformation, and SRR. A proof-of-concept is provided with two case studies involving CT, BLI, and MRI data of bone and kidney tumors in a mouse model. We show that local SRR-MRI is a computationally efficient complementary imaging modality for the precise characterization of tumor metastases, and that the method provides a feasible high-resolution alternative to conventional MRI.

  10. Practical considerations for high spatial and temporal resolution dynamic transmission electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Armstrong, Michael R. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States)], E-mail: armstrong30@llnl.gov; Boyden, Ken [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Browning, Nigel D. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Department of Chemical Engineering and Materials Science, University of California-Davis, One Shields Avenue, Davis, CA 95616 (United States); Campbell, Geoffrey H.; Colvin, Jeffrey D.; De Hope, William J.; Frank, Alan M. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Gibson, David J.; Hartemann, Fred [N Division, Physics and Advanced Technologies Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-280, Livermore, CA 94550 (United States); Kim, Judy S. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States); Department of Chemical Engineering and Materials Science, University of California-Davis, One Shields Avenue, Davis, CA 95616 (United States); King, Wayne E.; La Grange, Thomas B.; Pyke, Ben J.; Reed, Bryan W.; Shuttlesworth, Richard M.; Stuart, Brent C.; Torralva, Ben R. [Materials Science and Technology Division, Chemistry and Materials Science Directorate, Lawrence Livermore National Laboratory, P.O. Box 808, L-356, Livermore, CA 94550 (United States)

    2007-04-15

    Although recent years have seen significant advances in the spatial resolution possible in the transmission electron microscope (TEM), the temporal resolution of most microscopes is limited to video rate at best. This lack of temporal resolution means that our understanding of dynamic processes in materials is extremely limited. High temporal resolution in the TEM can be achieved, however, by replacing the normal thermionic or field emission source with a photoemission source. In this case the temporal resolution is limited only by the ability to create a short pulse of photoexcited electrons in the source, and this can be as short as a few femtoseconds. The operation of the photo-emission source and the control of the subsequent pulse of electrons (containing as many as 5x10{sup 7} electrons) create significant challenges for a standard microscope column that is designed to operate with a single electron in the column at any one time. In this paper, the generation and control of electron pulses in the TEM to obtain a temporal resolution <10{sup -6} s will be described and the effect of the pulse duration and current density on the spatial resolution of the instrument will be examined. The potential of these levels of temporal and spatial resolution for the study of dynamic materials processes will also be discussed.

  11. Practical considerations for high spatial and temporal resolution dynamic transmission electron microscopy

    International Nuclear Information System (INIS)

    Armstrong, Michael R.; Boyden, Ken; Browning, Nigel D.; Campbell, Geoffrey H.; Colvin, Jeffrey D.; De Hope, William J.; Frank, Alan M.; Gibson, David J.; Hartemann, Fred; Kim, Judy S.; King, Wayne E.; La Grange, Thomas B.; Pyke, Ben J.; Reed, Bryan W.; Shuttlesworth, Richard M.; Stuart, Brent C.; Torralva, Ben R.

    2007-01-01

    Although recent years have seen significant advances in the spatial resolution possible in the transmission electron microscope (TEM), the temporal resolution of most microscopes is limited to video rate at best. This lack of temporal resolution means that our understanding of dynamic processes in materials is extremely limited. High temporal resolution in the TEM can be achieved, however, by replacing the normal thermionic or field emission source with a photoemission source. In this case the temporal resolution is limited only by the ability to create a short pulse of photoexcited electrons in the source, and this can be as short as a few femtoseconds. The operation of the photo-emission source and the control of the subsequent pulse of electrons (containing as many as 5x10 7 electrons) create significant challenges for a standard microscope column that is designed to operate with a single electron in the column at any one time. In this paper, the generation and control of electron pulses in the TEM to obtain a temporal resolution -6 s will be described and the effect of the pulse duration and current density on the spatial resolution of the instrument will be examined. The potential of these levels of temporal and spatial resolution for the study of dynamic materials processes will also be discussed

  12. Saturated virtual fluorescence emission difference microscopy based on detector array

    Science.gov (United States)

    Liu, Shaocong; Sun, Shiyi; Kuang, Cuifang; Ge, Baoliang; Wang, Wensheng; Liu, Xu

    2017-07-01

    Virtual fluorescence emission difference microscopy (vFED) has been proposed recently to enhance the lateral resolution of confocal microscopy with a detector array, implemented by scanning a doughnut-shaped pattern. Theoretically, the resolution can be enhanced by around 1.3-fold compared with that in confocal microscopy. For further improvement of the resolving ability of vFED, a novel method is presented utilizing fluorescence saturation for super-resolution imaging, which we called saturated virtual fluorescence emission difference microscopy (svFED). With a point detector array, matched solid and hollow point spread functions (PSF) can be obtained by photon reassignment, and the difference results between them can be used to boost the transverse resolution. Results show that the diffraction barrier can be surpassed by at least 34% compared with that in vFED and the resolution is around 2-fold higher than that in confocal microscopy.

  13. Three-dimensional inversion recovery manganese-enhanced MRI of mouse brain using super-resolution reconstruction to visualize nuclei involved in higher brain function.

    Science.gov (United States)

    Poole, Dana S; Plenge, Esben; Poot, Dirk H J; Lakke, Egbert A J F; Niessen, Wiro J; Meijering, Erik; van der Weerd, Louise

    2014-07-01

    The visualization of activity in mouse brain using inversion recovery spin echo (IR-SE) manganese-enhanced MRI (MEMRI) provides unique contrast, but suffers from poor resolution in the slice-encoding direction. Super-resolution reconstruction (SRR) is a resolution-enhancing post-processing technique in which multiple low-resolution slice stacks are combined into a single volume of high isotropic resolution using computational methods. In this study, we investigated, first, whether SRR can improve the three-dimensional resolution of IR-SE MEMRI in the slice selection direction, whilst maintaining or improving the contrast-to-noise ratio of the two-dimensional slice stacks. Second, the contrast-to-noise ratio of SRR IR-SE MEMRI was compared with a conventional three-dimensional gradient echo (GE) acquisition. Quantitative experiments were performed on a phantom containing compartments of various manganese concentrations. The results showed that, with comparable scan times, the signal-to-noise ratio of three-dimensional GE acquisition is higher than that of SRR IR-SE MEMRI. However, the contrast-to-noise ratio between different compartments can be superior with SRR IR-SE MEMRI, depending on the chosen inversion time. In vivo experiments were performed in mice receiving manganese using an implanted osmotic pump. The results showed that SRR works well as a resolution-enhancing technique in IR-SE MEMRI experiments. In addition, the SRR image also shows a number of brain structures that are more clearly discernible from the surrounding tissues than in three-dimensional GE acquisition, including a number of nuclei with specific higher brain functions, such as memory, stress, anxiety and reward behavior. Copyright © 2014 John Wiley & Sons, Ltd.

  14. High-resolution noncontact AFM and Kelvin probe force microscopy investigations of self-assembled photovoltaic donor–acceptor dyads

    Directory of Open Access Journals (Sweden)

    Benjamin Grévin

    2016-06-01

    Full Text Available Self-assembled donor–acceptor dyads are used as model nanostructured heterojunctions for local investigations by noncontact atomic force microscopy (nc-AFM and Kelvin probe force microscopy (KPFM. With the aim to probe the photo-induced charge carrier generation, thin films deposited on transparent indium tin oxide substrates are investigated in dark conditions and upon illumination. The topographic and contact potential difference (CPD images taken under dark conditions are analysed in view of the results of complementary transmission electron microscopy (TEM experiments. After in situ annealing, it is shown that the dyads with longer donor blocks essentially lead to standing acceptor–donor lamellae, where the acceptor and donor groups are π-stacked in an edge-on configuration. The existence of strong CPD and surface photo-voltage (SPV contrasts shows that structural variations occur within the bulk of the edge-on stacks. SPV images with a very high lateral resolution are achieved, which allows for the resolution of local photo-charging contrasts at the scale of single edge-on lamella. This work paves the way for local investigations of the optoelectronic properties of donor–acceptor supramolecular architectures down to the elementary building block level.

  15. The mechanisms underlying the enhanced resolution of atomic force microscopy with functionalized tips

    International Nuclear Information System (INIS)

    Moll, Nikolaj; Gross, Leo; Mohn, Fabian; Curioni, Alessandro; Meyer, Gerhard

    2010-01-01

    By functionalizing the tip of an atomic force microscope (AFM) with a molecule or an atom that significantly contributes to the tip-sample interaction, the resolution can be dramatically enhanced. The interaction and therefore the resolution crucially depend on the chemical nature of the tip termination. Employing a tip functionalized with a CO molecule, atomic resolution of a pentacene molecule was recently demonstrated. In this work, the interaction between the CO tip and the pentacene imaged are studied with first principles calculations. The calculated frequency shifts compare very well with the experiment. The different energy contributions are analyzed and the Pauli energy is computed. We demonstrate that the source of the high resolution is Pauli repulsion, whereas van der Waals and electrostatic interactions only add a diffuse attractive background.

  16. Spatial and temporal resolution in cryo-electron microscopy : a scope for nano-chemistry

    NARCIS (Netherlands)

    Frederik, P.M.; Sommerdijk, N.A.J.M.

    2005-01-01

    Cryo-electron microscopy has evolved in an established approach to study the structure of bio-colloids. Recent developments in instrumentation and automation, often demanded by life sciences, made cryo-EM a general tool in colloid chemistry. Recently improved instrumentation for vitrification has

  17. High resolution magnetic force microscopy: instrumentation and application for recording media

    NARCIS (Netherlands)

    Porthun, Steffen; Porthun, S.

    This thesis describes aspects of the use of magnetic force microscopy for the study of magnetic recording media. The maximum achievable storage density in magnetic recording is limited by the magnetic reversal behaviour of the medium and by the stability of the written information. The shape and

  18. High resolution magnetic force microscopy using focused ion beam modified tips

    NARCIS (Netherlands)

    Phillips, G.N.; Siekman, Martin Herman; Abelmann, Leon; Lodder, J.C.

    2002-01-01

    Atomic force microscope tips coated by the thermal evaporation of a magnetic 30 nm thick Co film have been modified by focused ion beam milling with Ga+ ions to produce tips suitable for magnetic force microscopy. Such tips possess a planar magnetic element with high magnetic shape anisotropy, an

  19. Development of targeted STORM for super resolution imaging of biological samples using digital micro-mirror device

    Science.gov (United States)

    Valiya Peedikakkal, Liyana; Steventon, Victoria; Furley, Andrew; Cadby, Ashley J.

    2017-12-01

    We demonstrate a simple illumination system based on a digital mirror device which allows for fine control over the power and pattern of illumination. We apply this to localization microscopy (LM), specifically stochastic optical reconstruction microscopy (STORM). Using this targeted STORM, we were able to image a selected area of a labelled cell without causing photo-damage to the surrounding areas of the cell.

  20. Dual Super-Systolic Core for Real-Time Reconstructive Algorithms of High-Resolution Radar/SAR Imaging Systems

    Science.gov (United States)

    Atoche, Alejandro Castillo; Castillo, Javier Vázquez

    2012-01-01

    A high-speed dual super-systolic core for reconstructive signal processing (SP) operations consists of a double parallel systolic array (SA) machine in which each processing element of the array is also conceptualized as another SA in a bit-level fashion. In this study, we addressed the design of a high-speed dual super-systolic array (SSA) core for the enhancement/reconstruction of remote sensing (RS) imaging of radar/synthetic aperture radar (SAR) sensor systems. The selected reconstructive SP algorithms are efficiently transformed in their parallel representation and then, they are mapped into an efficient high performance embedded computing (HPEC) architecture in reconfigurable Xilinx field programmable gate array (FPGA) platforms. As an implementation test case, the proposed approach was aggregated in a HW/SW co-design scheme in order to solve the nonlinear ill-posed inverse problem of nonparametric estimation of the power spatial spectrum pattern (SSP) from a remotely sensed scene. We show how such dual SSA core, drastically reduces the computational load of complex RS regularization techniques achieving the required real-time operational mode. PMID:22736964