Sample records for diffraction resolution limit

  1. Statistical Limits to Super Resolution

    NASA Astrophysics Data System (ADS)

    Lucy, L. B.

    1992-08-01

    The limits imposed by photon statistics on the degree to which Rayleigh's resolution limit for diffraction-limited images can be surpassed by applying image restoration techniques are investigated. An approximate statistical theory is given for the number of detected photons required in the image of an unresolved pair of equal point sources in order that its information content allows in principle resolution by restoration. This theory is confirmed by numerical restoration experiments on synthetic images, and quantitative limits are presented for restoration of diffraction-limited images formed by slit and circular apertures.

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

    NASA Astrophysics Data System (ADS)

    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.

  3. Coherent x-ray diffraction imaging with nanofocused illumination.

    PubMed

    Schroer, C G; Boye, P; Feldkamp, J M; Patommel, J; Schropp, A; Schwab, A; Stephan, S; Burghammer, M; Schöder, S; Riekel, C

    2008-08-29

    Coherent x-ray diffraction imaging is an x-ray microscopy technique with the potential of reaching spatial resolutions well beyond the diffraction limits of x-ray microscopes based on optics. However, the available coherent dose at modern x-ray sources is limited, setting practical bounds on the spatial resolution of the technique. By focusing the available coherent flux onto the sample, the spatial resolution can be improved for radiation-hard specimens. A small gold particle (size <100 nm) was illuminated with a hard x-ray nanobeam (E=15.25 keV, beam dimensions approximately 100 x 100 nm2) and is reconstructed from its coherent diffraction pattern. A resolution of about 5 nm is achieved in 600 s exposure time.

  4. High-resolution scanning precession electron diffraction: Alignment and spatial resolution.

    PubMed

    Barnard, Jonathan S; Johnstone, Duncan N; Midgley, Paul A

    2017-03-01

    Methods are presented for aligning the pivot point of a precessing electron probe in the scanning transmission electron microscope (STEM) and for assessing the spatial resolution in scanning precession electron diffraction (SPED) experiments. The alignment procedure is performed entirely in diffraction mode, minimising probe wander within the bright-field (BF) convergent beam electron diffraction (CBED) disk and is used to obtain high spatial resolution SPED maps. Through analysis of the power spectra of virtual bright-field images extracted from the SPED data, the precession-induced blur was measured as a function of precession angle. At low precession angles, SPED spatial resolution was limited by electronic noise in the scan coils; whereas at high precession angles SPED spatial resolution was limited by tilt-induced two-fold astigmatism caused by the positive spherical aberration of the probe-forming lens. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Diffraction-Limited Plenoptic Imaging with Correlated Light

    NASA Astrophysics Data System (ADS)

    Pepe, Francesco V.; Di Lena, Francesco; Mazzilli, Aldo; Edrei, Eitan; Garuccio, Augusto; Scarcelli, Giuliano; D'Angelo, Milena

    2017-12-01

    Traditional optical imaging faces an unavoidable trade-off between resolution and depth of field (DOF). To increase resolution, high numerical apertures (NAs) are needed, but the associated large angular uncertainty results in a limited range of depths that can be put in sharp focus. Plenoptic imaging was introduced a few years ago to remedy this trade-off. To this aim, plenoptic imaging reconstructs the path of light rays from the lens to the sensor. However, the improvement offered by standard plenoptic imaging is practical and not fundamental: The increased DOF leads to a proportional reduction of the resolution well above the diffraction limit imposed by the lens NA. In this Letter, we demonstrate that correlation measurements enable pushing plenoptic imaging to its fundamental limits of both resolution and DOF. Namely, we demonstrate maintaining the imaging resolution at the diffraction limit while increasing the depth of field by a factor of 7. Our results represent the theoretical and experimental basis for the effective development of promising applications of plenoptic imaging.

  6. Diffraction-Limited Plenoptic Imaging with Correlated Light.

    PubMed

    Pepe, Francesco V; Di Lena, Francesco; Mazzilli, Aldo; Edrei, Eitan; Garuccio, Augusto; Scarcelli, Giuliano; D'Angelo, Milena

    2017-12-15

    Traditional optical imaging faces an unavoidable trade-off between resolution and depth of field (DOF). To increase resolution, high numerical apertures (NAs) are needed, but the associated large angular uncertainty results in a limited range of depths that can be put in sharp focus. Plenoptic imaging was introduced a few years ago to remedy this trade-off. To this aim, plenoptic imaging reconstructs the path of light rays from the lens to the sensor. However, the improvement offered by standard plenoptic imaging is practical and not fundamental: The increased DOF leads to a proportional reduction of the resolution well above the diffraction limit imposed by the lens NA. In this Letter, we demonstrate that correlation measurements enable pushing plenoptic imaging to its fundamental limits of both resolution and DOF. Namely, we demonstrate maintaining the imaging resolution at the diffraction limit while increasing the depth of field by a factor of 7. Our results represent the theoretical and experimental basis for the effective development of promising applications of plenoptic imaging.

  7. Breaking the acoustic diffraction barrier with localization optoacoustic tomography

    NASA Astrophysics Data System (ADS)

    Deán-Ben, X. Luís.; Razansky, Daniel

    2018-02-01

    Diffraction causes blurring of high-resolution features in images and has been traditionally associated to the resolution limit in light microscopy and other imaging modalities. The resolution of an imaging system can be generally assessed via its point spread function, corresponding to the image acquired from a point source. However, the precision in determining the position of an isolated source can greatly exceed the diffraction limit. By combining the estimated positions of multiple sources, localization-based imaging has resulted in groundbreaking methods such as super-resolution fluorescence optical microscopy and has also enabled ultrasound imaging of microvascular structures with unprecedented spatial resolution in deep tissues. Herein, we introduce localization optoacoustic tomography (LOT) and discuss on the prospects of using localization imaging principles in optoacoustic imaging. LOT was experimentally implemented by real-time imaging of flowing particles in 3D with a recently-developed volumetric optoacoustic tomography system. Provided the particles were separated by a distance larger than the diffraction-limited resolution, their individual locations could be accurately determined in each frame of the acquired image sequence and the localization image was formed by superimposing a set of points corresponding to the localized positions of the absorbers. The presented results demonstrate that LOT can significantly enhance the well-established advantages of optoacoustic imaging by breaking the acoustic diffraction barrier in deep tissues and mitigating artifacts due to limited-view tomographic acquisitions.

  8. Resolution enhancement by extrapolation of coherent diffraction images: a quantitative study on the limits and a numerical study of nonbinary and phase objects.

    PubMed

    Latychevskaia, T; Chushkin, Y; Fink, H-W

    2016-10-01

    In coherent diffractive imaging, the resolution of the reconstructed object is limited by the numerical aperture of the experimental setup. We present here a theoretical and numerical study for achieving super-resolution by postextrapolation of coherent diffraction images, such as diffraction patterns or holograms. We demonstrate that a diffraction pattern can unambiguously be extrapolated from only a fraction of the entire pattern and that the ratio of the extrapolated signal to the originally available signal is linearly proportional to the oversampling ratio. Although there could be in principle other methods to achieve extrapolation, we devote our discussion to employing iterative phase retrieval methods and demonstrate their limits. We present two numerical studies; namely, the extrapolation of diffraction patterns of nonbinary and that of phase objects together with a discussion of the optimal extrapolation procedure. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

  9. Macromolecular diffractive imaging using imperfect crystals

    PubMed Central

    Ayyer, Kartik; Yefanov, Oleksandr; Oberthür, Dominik; Roy-Chowdhury, Shatabdi; Galli, Lorenzo; Mariani, Valerio; Basu, Shibom; Coe, Jesse; Conrad, Chelsie E.; Fromme, Raimund; Schaffer, Alexander; Dörner, Katerina; James, Daniel; Kupitz, Christopher; Metz, Markus; Nelson, Garrett; Lourdu Xavier, Paulraj; Beyerlein, Kenneth R.; Schmidt, Marius; Sarrou, Iosifina; Spence, John C. H.; Weierstall, Uwe; White, Thomas A.; Yang, Jay-How; Zhao, Yun; Liang, Mengning; Aquila, Andrew; Hunter, Mark S.; Robinson, Joseph S.; Koglin, Jason E.; Boutet, Sébastien; Fromme, Petra; Barty, Anton; Chapman, Henry N.

    2016-01-01

    The three-dimensional structures of macromolecules and their complexes are predominantly elucidated by X-ray protein crystallography. A major limitation is access to high-quality crystals, to ensure X-ray diffraction extends to sufficiently large scattering angles and hence yields sufficiently high-resolution information that the crystal structure can be solved. The observation that crystals with shrunken unit-cell volumes and tighter macromolecular packing often produce higher-resolution Bragg peaks1,2 hints that crystallographic resolution for some macromolecules may be limited not by their heterogeneity but rather by a deviation of strict positional ordering of the crystalline lattice. Such displacements of molecules from the ideal lattice give rise to a continuous diffraction pattern, equal to the incoherent sum of diffraction from rigid single molecular complexes aligned along several discrete crystallographic orientations and hence with an increased information content3. Although such continuous diffraction patterns have long been observed—and are of interest as a source of information about the dynamics of proteins4 —they have not been used for structure determination. Here we show for crystals of the integral membrane protein complex photosystem II that lattice disorder increases the information content and the resolution of the diffraction pattern well beyond the 4.5 Å limit of measurable Bragg peaks, which allows us to directly phase5 the pattern. With the molecular envelope conventionally determined at 4.5 Å as a constraint, we then obtain a static image of the photosystem II dimer at 3.5 Å resolution. This result shows that continuous diffraction can be used to overcome long-supposed resolution limits of macromolecular crystallography, with a method that puts great value in commonly encountered imperfect crystals and opens up the possibility for model-free phasing6,7. PMID:26863980

  10. Super-resolution fluorescence microscopy by stepwise optical saturation

    PubMed Central

    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

  11. Printing colour at the optical diffraction limit.

    PubMed

    Kumar, Karthik; Duan, Huigao; Hegde, Ravi S; Koh, Samuel C W; Wei, Jennifer N; Yang, Joel K W

    2012-09-01

    The highest possible resolution for printed colour images is determined by the diffraction limit of visible light. To achieve this limit, individual colour elements (or pixels) with a pitch of 250 nm are required, translating into printed images at a resolution of ∼100,000 dots per inch (d.p.i.). However, methods for dispensing multiple colourants or fabricating structural colour through plasmonic structures have insufficient resolution and limited scalability. Here, we present a non-colourant method that achieves bright-field colour prints with resolutions up to the optical diffraction limit. Colour information is encoded in the dimensional parameters of metal nanostructures, so that tuning their plasmon resonance determines the colours of the individual pixels. Our colour-mapping strategy produces images with both sharp colour changes and fine tonal variations, is amenable to large-volume colour printing via nanoimprint lithography, and could be useful in making microimages for security, steganography, nanoscale optical filters and high-density spectrally encoded optical data storage.

  12. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Latychevskaia, Tatiana, E-mail: tatiana@physik.uzh.ch; Fink, Hans-Werner; Chushkin, Yuriy

    Coherent diffraction imaging is a high-resolution imaging technique whose potential can be greatly enhanced by applying the extrapolation method presented here. We demonstrate the enhancement in resolution of a non-periodical object reconstructed from an experimental X-ray diffraction record which contains about 10% missing information, including the pixels in the center of the diffraction pattern. A diffraction pattern is extrapolated beyond the detector area and as a result, the object is reconstructed at an enhanced resolution and better agreement with experimental amplitudes is achieved. The optimal parameters for the iterative routine and the limits of the extrapolation procedure are discussed.

  13. Improved Resolution Optical Time Stretch Imaging Based on High Efficiency In-Fiber Diffraction.

    PubMed

    Wang, Guoqing; Yan, Zhijun; Yang, Lei; Zhang, Lin; Wang, Chao

    2018-01-12

    Most overlooked challenges in ultrafast optical time stretch imaging (OTSI) are sacrificed spatial resolution and higher optical loss. These challenges are originated from optical diffraction devices used in OTSI, which encode image into spectra of ultrashort optical pulses. Conventional free-space diffraction gratings, as widely used in existing OTSI systems, suffer from several inherent drawbacks: limited diffraction efficiency in a non-Littrow configuration due to inherent zeroth-order reflection, high coupling loss between free-space gratings and optical fibers, bulky footprint, and more importantly, sacrificed imaging resolution due to non-full-aperture illumination for individual wavelengths. Here we report resolution-improved and diffraction-efficient OTSI using in-fiber diffraction for the first time to our knowledge. The key to overcome the existing challenges is a 45° tilted fiber grating (TFG), which serves as a compact in-fiber diffraction device offering improved diffraction efficiency (up to 97%), inherent compatibility with optical fibers, and improved imaging resolution owning to almost full-aperture illumination for all illumination wavelengths. 50 million frames per second imaging of fast moving object at 46 m/s with improved imaging resolution has been demonstrated. This conceptually new in-fiber diffraction design opens the way towards cost-effective, compact and high-resolution OTSI systems for image-based high-throughput detection and measurement.

  14. Speckle imaging for planetary research

    NASA Technical Reports Server (NTRS)

    Nisenson, P.; Goody, R.; Apt, J.; Papaliolios, C.

    1983-01-01

    The present study of speckle imaging technique effectiveness encompasses image reconstruction by means of a division algorithm for Fourier amplitudes, and the Knox-Thompson (1974) algorithm for Fourier phases. Results which have been obtained for Io, Titan, Pallas, Jupiter and Uranus indicate that spatial resolutions lower than the seeing limit by a factor of four are obtainable for objects brighter than Uranus. The resolutions obtained are well above the diffraction limit, due to inadequacies of the video camera employed. A photon-counting camera has been developed to overcome these difficulties, making possible the diffraction-limited resolution of objects as faint as Charon.

  15. Super-resolution differential interference contrast microscopy by structured illumination.

    PubMed

    Chen, Jianling; Xu, Yan; Lv, Xiaohua; Lai, Xiaomin; Zeng, Shaoqun

    2013-01-14

    We propose a structured illumination differential interference contrast (SI-DIC) microscopy, breaking the diffraction resolution limit of differential interference contrast (DIC) microscopy. SI-DIC extends the bandwidth of coherent transfer function of the DIC imaging system, thus the resolution is improved. With 0.8 numerical aperture condenser and objective, the reconstructed SI-DIC image of 53 nm polystyrene beads reveals lateral resolution of approximately 190 nm, doubling that of the conventional DIC image. We also demonstrate biological observations of label-free cells with improved spatial resolution. The SI-DIC microscopy can provide sub-diffraction resolution and high contrast images with marker-free specimens, and has the potential for achieving sub-diffraction resolution quantitative phase imaging.

  16. Breaking the acoustic diffraction limit via nonlinear effect and thermal confinement for potential deep-tissue high-resolution imaging

    PubMed Central

    Yuan, Baohong; Pei, Yanbo; Kandukuri, Jayanth

    2013-01-01

    Our recently developed ultrasound-switchable fluorescence (USF) imaging technique showed that it was feasible to conduct high-resolution fluorescence imaging in a centimeter-deep turbid medium. Because the spatial resolution of this technique highly depends on the ultrasound-induced temperature focal size (UTFS), minimization of UTFS becomes important for further improving the spatial resolution USF technique. In this study, we found that UTFS can be significantly reduced below the diffraction-limited acoustic intensity focal size via nonlinear acoustic effects and thermal confinement by appropriately controlling ultrasound power and exposure time, which can be potentially used for deep-tissue high-resolution imaging. PMID:23479498

  17. Femtosecond gas phase electron diffraction with MeV electrons.

    PubMed

    Yang, Jie; Guehr, Markus; Vecchione, Theodore; Robinson, Matthew S; Li, Renkai; Hartmann, Nick; Shen, Xiaozhe; Coffee, Ryan; Corbett, Jeff; Fry, Alan; Gaffney, Kelly; Gorkhover, Tais; Hast, Carsten; Jobe, Keith; Makasyuk, Igor; Reid, Alexander; Robinson, Joseph; Vetter, Sharon; Wang, Fenglin; Weathersby, Stephen; Yoneda, Charles; Wang, Xijie; Centurion, Martin

    2016-12-16

    We present results on ultrafast gas electron diffraction (UGED) experiments with femtosecond resolution using the MeV electron gun at SLAC National Accelerator Laboratory. UGED is a promising method to investigate molecular dynamics in the gas phase because electron pulses can probe the structure with a high spatial resolution. Until recently, however, it was not possible for UGED to reach the relevant timescale for the motion of the nuclei during a molecular reaction. Using MeV electron pulses has allowed us to overcome the main challenges in reaching femtosecond resolution, namely delivering short electron pulses on a gas target, overcoming the effect of velocity mismatch between pump laser pulses and the probe electron pulses, and maintaining a low timing jitter. At electron kinetic energies above 3 MeV, the velocity mismatch between laser and electron pulses becomes negligible. The relativistic electrons are also less susceptible to temporal broadening due to the Coulomb force. One of the challenges of diffraction with relativistic electrons is that the small de Broglie wavelength results in very small diffraction angles. In this paper we describe the new setup and its characterization, including capturing static diffraction patterns of molecules in the gas phase, finding time-zero with sub-picosecond accuracy and first time-resolved diffraction experiments. The new device can achieve a temporal resolution of 100 fs root-mean-square, and sub-angstrom spatial resolution. The collimation of the beam is sufficient to measure the diffraction pattern, and the transverse coherence is on the order of 2 nm. Currently, the temporal resolution is limited both by the pulse duration of the electron pulse on target and by the timing jitter, while the spatial resolution is limited by the average electron beam current and the signal-to-noise ratio of the detection system. We also discuss plans for improving both the temporal resolution and the spatial resolution.

  18. High-resolution ab initio three-dimensional x-ray diffraction microscopy

    DOE PAGES

    Chapman, Henry N.; Barty, Anton; Marchesini, Stefano; ...

    2006-01-01

    Coherent x-ray diffraction microscopy is a method of imaging nonperiodic isolated objects at resolutions limited, in principle, by only the wavelength and largest scattering angles recorded. We demonstrate x-ray diffraction imaging with high resolution in all three dimensions, as determined by a quantitative analysis of the reconstructed volume images. These images are retrieved from the three-dimensional diffraction data using no a priori knowledge about the shape or composition of the object, which has never before been demonstrated on a nonperiodic object. We also construct two-dimensional images of thick objects with greatly increased depth of focus (without loss of transverse spatialmore » resolution). These methods can be used to image biological and materials science samples at high resolution with x-ray undulator radiation and establishes the techniques to be used in atomic-resolution ultrafast imaging at x-ray free-electron laser sources.« less

  19. Enhancing resolution in coherent x-ray diffraction imaging.

    PubMed

    Noh, Do Young; Kim, Chan; Kim, Yoonhee; Song, Changyong

    2016-12-14

    Achieving a resolution near 1 nm is a critical issue in coherent x-ray diffraction imaging (CDI) for applications in materials and biology. Albeit with various advantages of CDI based on synchrotrons and newly developed x-ray free electron lasers, its applications would be limited without improving resolution well below 10 nm. Here, we review the issues and efforts in improving CDI resolution including various methods for resolution determination. Enhancing diffraction signal at large diffraction angles, with the aid of interference between neighboring strong scatterers or templates, is reviewed and discussed in terms of increasing signal-to-noise ratio. In addition, we discuss errors in image reconstruction algorithms-caused by the discreteness of the Fourier transformations involved-which degrade the spatial resolution, and suggest ways to correct them. We expect this review to be useful for applications of CDI in imaging weakly scattering soft matters using coherent x-ray sources including x-ray free electron lasers.

  20. Signal enhancement and Patterson-search phasing for high-spatial-resolution coherent X-ray diffraction imaging of biological objects.

    PubMed

    Takayama, Yuki; Maki-Yonekura, Saori; Oroguchi, Tomotaka; Nakasako, Masayoshi; Yonekura, Koji

    2015-01-28

    In this decade coherent X-ray diffraction imaging has been demonstrated to reveal internal structures of whole biological cells and organelles. However, the spatial resolution is limited to several tens of nanometers due to the poor scattering power of biological samples. The challenge is to recover correct phase information from experimental diffraction patterns that have a low signal-to-noise ratio and unmeasurable lowest-resolution data. Here, we propose a method to extend spatial resolution by enhancing diffraction signals and by robust phasing. The weak diffraction signals from biological objects are enhanced by interference with strong waves from dispersed colloidal gold particles. The positions of the gold particles determined by Patterson analysis serve as the initial phase, and this dramatically improves reliability and convergence of image reconstruction by iterative phase retrieval. A set of calculations based on current experiments demonstrates that resolution is improved by a factor of two or more.

  1. Signal enhancement and Patterson-search phasing for high-spatial-resolution coherent X-ray diffraction imaging of biological objects

    PubMed Central

    Takayama, Yuki; Maki-Yonekura, Saori; Oroguchi, Tomotaka; Nakasako, Masayoshi; Yonekura, Koji

    2015-01-01

    In this decade coherent X-ray diffraction imaging has been demonstrated to reveal internal structures of whole biological cells and organelles. However, the spatial resolution is limited to several tens of nanometers due to the poor scattering power of biological samples. The challenge is to recover correct phase information from experimental diffraction patterns that have a low signal-to-noise ratio and unmeasurable lowest-resolution data. Here, we propose a method to extend spatial resolution by enhancing diffraction signals and by robust phasing. The weak diffraction signals from biological objects are enhanced by interference with strong waves from dispersed colloidal gold particles. The positions of the gold particles determined by Patterson analysis serve as the initial phase, and this dramatically improves reliability and convergence of image reconstruction by iterative phase retrieval. A set of calculations based on current experiments demonstrates that resolution is improved by a factor of two or more. PMID:25627480

  2. Weak data do not make a free lunch, only a cheap meal

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Luo, Zhipu; Rajashankar, Kanagalaghatta; Dauter, Zbigniew, E-mail: dauter@anl.gov

    2014-02-01

    Refinement and analysis of four structures with various data resolution cutoffs suggests that at present there are no reliable criteria for judging the diffraction data resolution limit and the condition I/σ(I) = 2.0 is reasonable. However, extending the limit by about 0.2 Å beyond the resolution defined by this threshold does not deteriorate the quality of refined structures and in some cases may be beneficial. Four data sets were processed at resolutions significantly exceeding the criteria traditionally used for estimating the diffraction data resolution limit. The analysis of these data and the corresponding model-quality indicators suggests that the criteria ofmore » resolution limits widely adopted in the past may be somewhat conservative. Various parameters, such as R{sub merge} and I/σ(I), optical resolution and the correlation coefficients CC{sub 1/2} and CC*, can be used for judging the internal data quality, whereas the reliability factors R and R{sub free} as well as the maximum-likelihood target values and real-space map correlation coefficients can be used to estimate the agreement between the data and the refined model. However, none of these criteria provide a reliable estimate of the data resolution cutoff limit. The analysis suggests that extension of the maximum resolution by about 0.2 Å beyond the currently adopted limit where the I/σ(I) value drops to 2.0 does not degrade the quality of the refined structural models, but may sometimes be advantageous. Such an extension may be particularly beneficial for significantly anisotropic diffraction. Extension of the maximum resolution at the stage of data collection and structure refinement is cheap in terms of the required effort and is definitely more advisable than accepting a too conservative resolution cutoff, which is unfortunately quite frequent among the crystal structures deposited in the Protein Data Bank.« less

  3. Breaking resolution limits in ultrafast electron diffraction and microscopy.

    PubMed

    Baum, Peter; Zewail, Ahmed H

    2006-10-31

    Ultrafast electron microscopy and diffraction are powerful techniques for the study of the time-resolved structures of molecules, materials, and biological systems. Central to these approaches is the use of ultrafast coherent electron packets. The electron pulses typically have an energy of 30 keV for diffraction and 100-200 keV for microscopy, corresponding to speeds of 33-70% of the speed of light. Although the spatial resolution can reach the atomic scale, the temporal resolution is limited by the pulse width and by the difference in group velocities of electrons and the light used to initiate the dynamical change. In this contribution, we introduce the concept of tilted optical pulses into diffraction and imaging techniques and demonstrate the methodology experimentally. These advances allow us to reach limits of time resolution down to regimes of a few femtoseconds and, possibly, attoseconds. With tilted pulses, every part of the sample is excited at precisely the same time as when the electrons arrive at the specimen. Here, this approach is demonstrated for the most unfavorable case of ultrafast crystallography. We also present a method for measuring the duration of electron packets by autocorrelating electron pulses in free space and without streaking, and we discuss the potential of tilting the electron pulses themselves for applications in domains involving nuclear and electron motions.

  4. Dynamic microscale temperature gradient in a gold nanorod solution measured by diffraction-limited nanothermometry

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, Chengmingyue; Gan, Xiaosong; Li, Xiangping

    2015-09-21

    We quantify the dynamic microscale temperature gradient in a gold nanorod solution using quantum-dot-based microscopic fluorescence nanothermometry. By incorporating CdSe quantum dots into the solution as a nanothermometer, precise temperature mapping with diffraction-limited spatial resolution and sub-degree temperature resolution is achieved. The acquired data on heat generation and dissipation show an excellent agreement with theoretical simulations. This work reveals an effective approach for noninvasive temperature regulation with localized nanoheaters in microfluidic environment.

  5. Structured illumination multimodal 3D-resolved quantitative phase and fluorescence sub-diffraction microscopy

    PubMed Central

    Chowdhury, Shwetadwip; Eldridge, Will J.; Wax, Adam; Izatt, Joseph A.

    2017-01-01

    Sub-diffraction resolution imaging has played a pivotal role in biological research by visualizing key, but previously unresolvable, sub-cellular structures. Unfortunately, applications of far-field sub-diffraction resolution are currently divided between fluorescent and coherent-diffraction regimes, and a multimodal sub-diffraction technique that bridges this gap has not yet been demonstrated. Here we report that structured illumination (SI) allows multimodal sub-diffraction imaging of both coherent quantitative-phase (QP) and fluorescence. Due to SI’s conventionally fluorescent applications, we first demonstrate the principle of SI-enabled three-dimensional (3D) QP sub-diffraction imaging with calibration microspheres. Image analysis confirmed enhanced lateral and axial resolutions over diffraction-limited QP imaging, and established striking parallels between coherent SI and conventional optical diffraction tomography. We next introduce an optical system utilizing SI to achieve 3D sub-diffraction, multimodal QP/fluorescent visualization of A549 biological cells fluorescently tagged for F-actin. Our results suggest that SI has a unique utility in studying biological phenomena with significant molecular, biophysical, and biochemical components. PMID:28663887

  6. High-resolution x-ray diffraction microscopy of specifically labeled yeast cells

    PubMed Central

    Nelson, Johanna; Huang, Xiaojing; Steinbrener, Jan; Shapiro, David; Kirz, Janos; Marchesini, Stefano; Neiman, Aaron M.; Turner, Joshua J.; Jacobsen, Chris

    2010-01-01

    X-ray diffraction microscopy complements other x-ray microscopy methods by being free of lens-imposed radiation dose and resolution limits, and it allows for high-resolution imaging of biological specimens too thick to be viewed by electron microscopy. We report here the highest resolution (11–13 nm) x-ray diffraction micrograph of biological specimens, and a demonstration of molecular-specific gold labeling at different depths within cells via through-focus propagation of the reconstructed wavefield. The lectin concanavalin A conjugated to colloidal gold particles was used to label the α-mannan sugar in the cell wall of the yeast Saccharomyces cerevisiae. Cells were plunge-frozen in liquid ethane and freeze-dried, after which they were imaged whole using x-ray diffraction microscopy at 750 eV photon energy. PMID:20368463

  7. High-resolution x-ray diffraction microscopy of specifically labeled yeast cells

    DOE PAGES

    Nelson, Johanna; Huang, Xiaojing; Steinbrener, Jan; ...

    2010-04-20

    X-ray diffraction microscopy complements other x-ray microscopy methods by being free of lens-imposed radiation dose and resolution limits, and it allows for high-resolution imaging of biological specimens too thick to be viewed by electron microscopy. We report here the highest resolution (11-13 nm) x-ray diffraction micrograph of biological specimens, and a demonstration of molecular-specific gold labeling at different depths within cells via through-focus propagation of the reconstructed wavefield. The lectin concanavalin A conjugated to colloidal gold particles was used to label the α-mannan sugar in the cell wall of the yeast Saccharomyces cerevisiae. Cells were plunge-frozen in liquid ethane andmore » freeze-dried, after which they were imaged whole using x-ray diffraction microscopy at 750 eV photon energy.« less

  8. 15x optical zoom and extreme optical image stabilisation: diffraction limited integral field spectroscopy with the Oxford SWIFT spectrograph

    NASA Astrophysics Data System (ADS)

    Tecza, Matthias; Thatte, Niranjan; Clarke, Fraser; Lynn, James; Freeman, David; Roberts, Jennifer; Dekany, Richard

    2012-09-01

    When commissioned in November 2008 at the Palomar 200 inch Hale Telescope, the Oxford SWIFT I and z band integral field spectrograph, fed by the adaptive optics system PALAO, provided a wide (3×) range of spatial resolutions: three plate scales of 235 mas, 160 mas, and 80 mas per spaxel over a contiguous field-of-view of 89×44 pixels. Depending on observing conditions and guide star brightness we can choose a seeing limited scale of 235 mas per spaxel, or 160 mas and 80 mas per spaxel for very bright guide star AO with substantial increase of enclosed energy. Over the last two years PALAO was upgraded to PALM-3000: an extreme, high-order adaptive optics system with two deformable mirrors with more than 3000 actuators, promising diffraction limited performance in SWIFT's wavelength range. In order to take advantage of this increased spatial resolution we upgraded SWIFT with new pre-optics allowing us to spatially Nyquist sample the diffraction limited PALM-3000 point spread function with 16 mas resolution, reducing the spaxel scale by another factor of 5×. We designed, manufactured, integrated and tested the new pre-optics in the first half of 2011 and commissioned it in December 2011. Here we present the opto-mechanical design and assembly of the new scale changing optics, as well as laboratory and on-sky commissioning results. In optimal observing conditions we achieve substantial Strehl ratios, delivering the near diffraction limited spatial resolution in the I and z bands.

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

    PubMed

    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.

  10. X-ray nanoprobes and diffraction-limited storage rings: opportunities and challenges of fluorescence tomography of biological specimens

    PubMed Central

    de Jonge, Martin D.; Ryan, Christopher G.; Jacobsen, Chris J.

    2014-01-01

    X-ray nanoprobes require coherent illumination to achieve optic-limited resolution, and so will benefit directly from diffraction-limited storage rings. Here, the example of high-resolution X-ray fluorescence tomography is focused on as one of the most voracious demanders of coherent photons, since the detected signal is only a small fraction of the incident flux. Alternative schemes are considered for beam delivery, sample scanning and detectors. One must consider as well the steps before and after the X-ray experiment: sample preparation and examination conditions, and analysis complexity due to minimum dose requirements and self-absorption. By understanding the requirements and opportunities for nanoscale fluorescence tomography, one gains insight into the R&D challenges in optics and instrumentation needed to fully exploit the source advances that diffraction-limited storage rings offer. PMID:25177992

  11. Breaking resolution limits in ultrafast electron diffraction and microscopy

    PubMed Central

    Baum, Peter; Zewail, Ahmed H.

    2006-01-01

    Ultrafast electron microscopy and diffraction are powerful techniques for the study of the time-resolved structures of molecules, materials, and biological systems. Central to these approaches is the use of ultrafast coherent electron packets. The electron pulses typically have an energy of 30 keV for diffraction and 100–200 keV for microscopy, corresponding to speeds of 33–70% of the speed of light. Although the spatial resolution can reach the atomic scale, the temporal resolution is limited by the pulse width and by the difference in group velocities of electrons and the light used to initiate the dynamical change. In this contribution, we introduce the concept of tilted optical pulses into diffraction and imaging techniques and demonstrate the methodology experimentally. These advances allow us to reach limits of time resolution down to regimes of a few femtoseconds and, possibly, attoseconds. With tilted pulses, every part of the sample is excited at precisely the same time as when the electrons arrive at the specimen. Here, this approach is demonstrated for the most unfavorable case of ultrafast crystallography. We also present a method for measuring the duration of electron packets by autocorrelating electron pulses in free space and without streaking, and we discuss the potential of tilting the electron pulses themselves for applications in domains involving nuclear and electron motions. PMID:17056711

  12. Super-Resolution Scanning Laser Microscopy Based on Virtually Structured Detection

    PubMed Central

    Zhi, Yanan; Wang, Benquan; Yao, Xincheng

    2016-01-01

    Light microscopy plays a key role in biological studies and medical diagnosis. The spatial resolution of conventional optical microscopes is limited to approximately half the wavelength of the illumination light as a result of the diffraction limit. Several approaches—including confocal microscopy, stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy, and structured illumination microscopy—have been established to achieve super-resolution imaging. However, none of these methods is suitable for the super-resolution ophthalmoscopy of retinal structures because of laser safety issues and inevitable eye movements. We recently experimentally validated virtually structured detection (VSD) as an alternative strategy to extend the diffraction limit. Without the complexity of structured illumination, VSD provides an easy, low-cost, and phase artifact–free strategy to achieve super-resolution in scanning laser microscopy. In this article we summarize the basic principles of the VSD method, review our demonstrated single-point and line-scan super-resolution systems, and discuss both technical challenges and the potential of VSD-based instrumentation for super-resolution ophthalmoscopy of the retina. PMID:27480461

  13. Spread spectrum phase modulation for coherent X-ray diffraction imaging.

    PubMed

    Zhang, Xuesong; Jiang, Jing; Xiangli, Bin; Arce, Gonzalo R

    2015-09-21

    High dynamic range, phase ambiguity and radiation limited resolution are three challenging issues in coherent X-ray diffraction imaging (CXDI), which limit the achievable imaging resolution. This paper proposes a spread spectrum phase modulation (SSPM) method to address the aforementioned problems in a single strobe. The requirements on phase modulator parameters are presented, and a practical implementation of SSPM is discussed via ray optics analysis. Numerical experiments demonstrate the performance of SSPM under the constraint of available X-ray optics fabrication accuracy, showing its potential to real CXDI applications.

  14. Low-dose, high-resolution and high-efficiency ptychography at STXM beamline of SSRF

    NASA Astrophysics Data System (ADS)

    Xu, Zijian; Wang, Chunpeng; Liu, Haigang; Tao, Xulei; Tai, Renzhong

    2017-06-01

    Ptychography is a diffraction-based X-ray microscopy method that can image extended samples quantitatively while remove the resolution limit imposed by image-forming optical elements. As a natural extension of scanning transmission X-ray microscopy (STXM) imaging method, we developed soft X-ray ptychographic coherent diffraction imaging (PCDI) method at the STXM endstation of BL08U beamline of Shanghai Synchrotron Radiation Facility. Compared to the traditional STXM imaging, the new PCDI method has resulted in significantly lower dose, higher resolution and higher efficiency imaging in our platform. In the demonstration experiments shown here, a spatial resolution of sub-10 nm was obtained for a gold nanowires sample, which is much better than the limit resolution 30 nm of the STXM method, while the radiation dose is only 1/12 of STXM.

  15. Diffraction-limited real-time terahertz imaging by optical frequency up-conversion in a DAST crystal.

    PubMed

    Fan, Shuzhen; Qi, Feng; Notake, Takashi; Nawata, Kouji; Takida, Yuma; Matsukawa, Takeshi; Minamide, Hiroaki

    2015-03-23

    Real-time terahertz (THz) wave imaging has wide applications in areas such as security, industry, biology, medicine, pharmacy, and the arts. This report describes real-time room-temperature THz imaging by nonlinear optical frequency up-conversion in an organic 4-dimethylamino-N'-methyl-4'-stilbazolium tosylate (DAST) crystal, with high resolution reaching the diffraction limit. THz-wave images were converted to the near infrared region and then captured using an InGaAs camera in a tandem imaging system. The resolution of the imaging system was analyzed. Diffraction and interference of THz wave were observed in the experiments. Videos are supplied to show the interference pattern variation that occurs with sample moving and tilting.

  16. Imaging whole Escherichia coli bacteria by using single-particle x-ray diffraction

    NASA Astrophysics Data System (ADS)

    Miao, Jianwei; Hodgson, Keith O.; Ishikawa, Tetsuya; Larabell, Carolyn A.; Legros, Mark A.; Nishino, Yoshinori

    2003-01-01

    We report the first experimental recording, to our knowledge, of the diffraction pattern from intact Escherichia coli bacteria using coherent x-rays with a wavelength of 2 Å. By using the oversampling phasing method, a real space image at a resolution of 30 nm was directly reconstructed from the diffraction pattern. An R factor used for characterizing the quality of the reconstruction was in the range of 5%, which demonstrated the reliability of the reconstruction process. The distribution of proteins inside the bacteria labeled with manganese oxide has been identified and this distribution confirmed by fluorescence microscopy images. Compared with lens-based microscopy, this diffraction-based imaging approach can examine thicker samples, such as whole cultured cells, in three dimensions with resolution limited only by radiation damage. Looking forward, the successful recording and reconstruction of diffraction patterns from biological samples reported here represent an important step toward the potential of imaging single biomolecules at near-atomic resolution by combining single-particle diffraction with x-ray free electron lasers.

  17. Soft X-Ray Diffraction Microscopy of a Frozen Hydrated Yeast Cell

    DOE PAGES

    Huang, Xiaojing; Nelson, Johanna; Kirz, Janos; ...

    2009-11-01

    We report the first image of an intact, frozen hydrated eukaryotic cell using x-ray diffraction microscopy, or coherent x-ray diffraction imaging. By plunge freezing the specimen in liquid ethane and maintaining it below -170 °C, artifacts due to dehydration, ice crystallization, and radiation damage are greatly reduced. In this example, coherent diffraction data using 520 eV x rays were recorded and reconstructed to reveal a budding yeast cell at a resolution better than 25 nm. This demonstration represents an important step towards high resolution imaging of cells in their natural, hydrated state, without limitations imposed by x-ray optics.

  18. Synthetic light-needle photoacoustic microscopy for extended depth of field (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Yang, Jiamiao; Gong, Lei; Xu, Xiao; Hai, Pengfei; Suzuki, Yuta; Wang, Lihong V.

    2017-03-01

    Photoacoustic microscopy (PAM) has been extensively applied in biomedical study because of its ability to visualize tissue morphology and physiology in vivo in three dimensions (3D). However, conventional PAM suffers from a rapidly decreasing resolution away from the focal plane because of the limited depth of focus of an objective lens, which deteriorates the volumetric imaging quality inevitably. Here, we propose a novel method to synthesize an ultra-long light needle to extend a microscope's depth of focus beyond its physical limitations with wavefront engineering method. Furthermore, it enables an improved lateral resolution that exceeds the diffraction limit of the objective lens. The virtual light needle can be flexibly synthesized anywhere throughout the imaging volume without mechanical scanning. Benefiting from these advantages, we developed a synthetic light needle photoacoustic microscopy (SLN-PAM) to achieve an extended depth of field (DOF), sub-diffraction and motionless volumetric imaging. The DOF of our SLN-PAM system is up to 1800 µm, more than 30-fold improvement over that gained by conventional PAM. Our system also achieves the lateral resolution of 1.8 µm (characterized at 532 nm and 0.1 NA objective), about 50% higher than the Rayleigh diffraction limit. Its superior imaging performance was demonstrated by 3D imaging of both non-biological and biological samples. This extended DOF, sub-diffraction and motionless 3D PAM will open up new opportunities for potential biomedical applications.

  19. Bayesian superresolution

    NASA Astrophysics Data System (ADS)

    Isakson, Steve Wesley

    2001-12-01

    Well-known principles of physics explain why resolution restrictions occur in images produced by optical diffraction-limited systems. The limitations involved are present in all diffraction-limited imaging systems, including acoustical and microwave. In most circumstances, however, prior knowledge about the object and the imaging system can lead to resolution improvements. In this dissertation I outline a method to incorporate prior information into the process of reconstructing images to superresolve the object beyond the above limitations. This dissertation research develops the details of this methodology. The approach can provide the most-probable global solution employing a finite number of steps in both far-field and near-field images. In addition, in order to overcome the effects of noise present in any imaging system, this technique provides a weighted image that quantifies the likelihood of various imaging solutions. By utilizing Bayesian probability, the procedure is capable of incorporating prior information about both the object and the noise to overcome the resolution limitation present in many imaging systems. Finally I will present an imaging system capable of detecting the evanescent waves missing from far-field systems, thus improving the resolution further.

  20. Scanning digital lithography providing high speed large area patterning with diffraction limited sub-micron resolution

    NASA Astrophysics Data System (ADS)

    Wen, Sy-Bor; Bhaskar, Arun; Zhang, Hongjie

    2018-07-01

    A scanning digital lithography system using computer controlled digital spatial light modulator, spatial filter, infinity correct optical microscope and high precision translation stage is proposed and examined. Through utilizing the spatial filter to limit orders of diffraction modes for light delivered from the spatial light modulator, we are able to achieve diffraction limited deep submicron spatial resolution with the scanning digital lithography system by using standard one inch level optical components with reasonable prices. Raster scanning of this scanning digital lithography system using a high speed high precision x-y translation stage and piezo mount to real time adjust the focal position of objective lens allows us to achieve large area sub-micron resolved patterning with high speed (compared with e-beam lithography). It is determined in this study that to achieve high quality stitching of lithography patterns with raster scanning, a high-resolution rotation stage will be required to ensure the x and y directions of the projected pattern are in the same x and y translation directions of the nanometer precision x-y translation stage.

  1. High-resolution three-dimensional partially coherent diffraction imaging.

    PubMed

    Clark, J N; Huang, X; Harder, R; Robinson, I K

    2012-01-01

    The wave properties of light, particularly its coherence, are responsible for interference effects, which can be exploited in powerful imaging applications. Coherent diffractive imaging relies heavily on coherence and has recently experienced rapid growth. Coherent diffractive imaging recovers an object from its diffraction pattern by computational phasing with the potential of wavelength-limited resolution. Diminished coherence results in reconstructions that suffer from artefacts or fail completely. Here we demonstrate ab initio phasing of partially coherent diffraction patterns in three dimensions, while simultaneously determining the coherence properties of the illuminating wavefield. Both the dramatic improvements in image interpretability and the three-dimensional evaluation of the coherence will have broad implications for quantitative imaging of nanostructures and wavefield characterization with X-rays and electrons.

  2. Role of coherence in microsphere-assisted nanoscopy

    NASA Astrophysics Data System (ADS)

    Perrin, Stephane; Lecler, Sylvain; Leong-Hoi, Audrey; Montgomery, Paul C.

    2017-06-01

    The loss of the information, due to the diffraction and the evanescent waves, limits the resolving power of classical optical microscopy. In air, the lateral resolution of an optical microscope can approximated at half of the wavelength using a low-coherence illumination. Recently, several methods have been developed in order to overcome this limitation and, in 2011, a new far-field and full-field imaging technique was proposed where a sub-diffraction-limit resolution has been achieved using a transparent microsphere. In this article, the phenomenon of super-resolution using microsphere-assisted microscopy is analysed through rigorous electro-magnetic simulations. The performances of the imaging technique are estimated as function of optical and geometrical parameters. Furthermore, the role of coherence is introduced through the temporal coherence of the light source and the phase response of the object.

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

    PubMed

    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.

  4. Defect of focus in two-line resolution with Hanning amplitude filters

    NASA Astrophysics Data System (ADS)

    Karunasagar, D.; Bhikshamaiah, G.; Keshavulu Goud, M.; Lacha Goud, S.

    In the presence of defocusing the modified Sparrow limits of resolution for two-line objects have been investigated for a diffraction-limited coherent optical system apodized by generalized Hanning amplitude filters. These limits have been studied as a function of different parameters such as intensity ratio, the order of the filter for various amounts of apodization parameter. Results reveal that in some situations the defocusing is effective in enhancing the resolution of an optical system.

  5. High resolution telescope

    DOEpatents

    Massie, Norbert A.; Oster, Yale

    1992-01-01

    A large effective-aperture, low-cost optical telescope with diffraction-limited resolution enables ground-based observation of near-earth space objects. The telescope has a non-redundant, thinned-aperture array in a center-mount, single-structure space frame. It employs speckle interferometric imaging to achieve diffraction-limited resolution. The signal-to-noise ratio problem is mitigated by moving the wavelength of operation to the near-IR, and the image is sensed by a Silicon CCD. The steerable, single-structure array presents a constant pupil. The center-mount, radar-like mount enables low-earth orbit space objects to be tracked as well as increases stiffness of the space frame. In the preferred embodiment, the array has elemental telescopes with subaperture of 2.1 m in a circle-of-nine configuration. The telescope array has an effective aperture of 12 m which provides a diffraction-limited resolution of 0.02 arc seconds. Pathlength matching of the telescope array is maintained by an electro-optical system employing laser metrology. Speckle imaging relaxes pathlength matching tolerance by one order of magnitude as compared to phased arrays. Many features of the telescope contribute to substantial reduction in costs. These include eliminating the conventional protective dome and reducing on-site construction activites. The cost of the telescope scales with the first power of the aperture rather than its third power as in conventional telescopes.

  6. Weak data do not make a free lunch, only a cheap meal

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Luo, Zhipu; Rajashankar, Kanagalaghatta; Dauter, Zbigniew

    2014-01-17

    Four data sets were processed at resolutions significantly exceeding the criteria traditionally used for estimating the diffraction data resolution limit. The analysis of these data and the corresponding model-quality indicators suggests that the criteria of resolution limits widely adopted in the past may be somewhat conservative. Various parameters, such asR mergeandI/σ(I), optical resolution and the correlation coefficients CC 1/2and CC*, can be used for judging the internal data quality, whereas the reliability factorsRandR freeas well as the maximum-likelihood target values and real-space map correlation coefficients can be used to estimate the agreement between the data and the refined model. However,more » none of these criteria provide a reliable estimate of the data resolution cutoff limit. The analysis suggests that extension of the maximum resolution by about 0.2 Å beyond the currently adopted limit where theI/σ(I) value drops to 2.0 does not degrade the quality of the refined structural models, but may sometimes be advantageous. Such an extension may be particularly beneficial for significantly anisotropic diffraction. Extension of the maximum resolution at the stage of data collection and structure refinement is cheap in terms of the required effort and is definitely more advisable than accepting a too conservative resolution cutoff, which is unfortunately quite frequent among the crystal structures deposited in the Protein Data Bank.« less

  7. X-ray diffraction microscopy on frozen hydrated specimens

    NASA Astrophysics Data System (ADS)

    Nelson, Johanna

    X-rays are excellent for imaging thick samples at high resolution because of their large penetration depth compared to electrons and their short wavelength relative to visible light. To image biological material, the absorption contrast of soft X-rays, especially between the carbon and oxygen K-shell absorption edges, can be utilized to give high contrast, high resolution images without the need for stains or labels. Because of radiation damage and the desire for high resolution tomography, live cell imaging is not feasible. However, cells can be frozen in vitrified ice, which reduces the effect of radiation damage while maintaining their natural hydrated state. X-ray diffraction microscopy (XDM) is an imaging technique which eliminates the limitations imposed by current focusing optics simply by removing them entirely. Far-field coherent diffraction intensity patterns are collected on a pixelated detector allowing every scattered photon to be collected within the limits of the detector's efficiency and physical size. An iterative computer algorithm is then used to invert the diffraction intensity into a real space image with both absorption and phase information. This technique transfers the emphasis away from fabrication and alignment of optics, and towards data processing. We have used this method to image a pair of freeze-dried, immuno-labeled yeast cells to the highest resolution (13 nm) yet obtained for a whole eukaryotic cell. We discuss successes and challenges in working with frozen hydrated specimens and efforts aimed at high resolution imaging of vitrified eukaryotic cells in 3D.

  8. Numerical investigations of the potential for laser focus sensors in micrometrology

    NASA Astrophysics Data System (ADS)

    Bischoff, Jörg; Mastylo, Rostyslav; Manske, Eberhard

    2017-06-01

    Laser focus sensors (LFS)1 attached to a scanning nano-positioning and measuring machine (NPMM) enable near diffraction limit resolution with very large measuring areas up to 200 x 200 mm1. Further extensions are planned to address wafer sizes of 8 inch and beyond. Thus, they are preferably suited for micro-metrology on large wafers. On the other hand, the minimum lateral features in state-of-the-art semiconductor industry are as small as a few nanometer and therefore far beyond the resolution limits of classical optics. New techniques such as OCD or ODP3,4 a.k.a. as scatterometry have helped to overcome these constraints considerably. However, scatterometry relies on regular patterns and therefore, the measurements have to be performed on special reference gratings or boxes rather than in-die. Consequently, there is a gap between measurement and the actual structure of interest which becomes more and more an issues with shrinking feature sizes. On the other hand, near-field approaches would also allow to extent the resolution limit greatly5 but they require very challenging controls to keep the working distance small enough to stay within the near field zone. Therefore, the feasibility and the limits of a LFS scanner system have been investigated theoretically. Based on simulations of laser focus sensor scanning across simple topographies, it was found that there is potential to overcome the diffraction limitations to some extent by means of vicinity interference effects caused by the optical interaction of adjacent topography features. We think that it might be well possible to reconstruct the diffracting profile by means of rigorous diffraction simulation based on a thorough model of the laser focus sensor optics in combination with topography diffraction 6 in a similar way as applied in OCD. The difference lies in the kind of signal itself which has to be modeled. While standard OCD is based on spectra, LFS utilizes height scan signals. Simulation results are presented for different types of topographies (dense vs. sparse, regular vs. single) with lateral features near and beyond the classical resolution limit. Moreover, the influence of topography height on the detectability is investigated. To this end, several sensor principles and polarization setups are considered such as a dual color pin hole sensor and a Foucault knife sensor. It is shown that resolution beyond the Abbe or Rayleigh limit is possible even with "classical" optical setups when combining measurements with sophisticated profile retrieval techniques and some a-priori knowledge. Finally, measurement uncertainties are derived based on perturbation simulations according to the method presented in 7.

  9. Microscale optical cryptography using a subdiffraction-limit optical key

    NASA Astrophysics Data System (ADS)

    Ogura, Yusuke; Aino, Masahiko; Tanida, Jun

    2018-04-01

    We present microscale optical cryptography using a subdiffraction-limit optical pattern, which is finer than the diffraction-limit size of the decrypting optical system, as a key and a substrate with a reflectance distribution as an encrypted image. Because of the subdiffraction-limit spatial coding, this method enables us to construct a secret image with the diffraction-limit resolution. Simulation and experimental results demonstrate, both qualitatively and quantitatively, that the secret image becomes recognizable when and only when the substrate is illuminated with the designed key pattern.

  10. High resolution observations of low contrast phenomena from an Advanced Geosynchronous Platform (AGP)

    NASA Technical Reports Server (NTRS)

    Maxwell, M. S.

    1984-01-01

    Present technology allows radiometric monitoring of the Earth, ocean and atmosphere from a geosynchronous platform with good spatial, spectral and temporal resolution. The proposed system could provide a capability for multispectral remote sensing with a 50 m nadir spatial resolution in the visible bands, 250 m in the 4 micron band and 1 km in the 11 micron thermal infrared band. The diffraction limited telescope has a 1 m aperture, a 10 m focal length (with a shorter focal length in the infrared) and linear and area arrays of detectors. The diffraction limited resolution applies to scenes of any brightness but for a dark low contrast scenes, the good signal to noise ratio of the system contribute to the observation capability. The capabilities of the AGP system are assessed for quantitative observations of ocean scenes. Instrument and ground system configuration are presented and projected sensor capabilities are analyzed.

  11. Prospects of high-resolution resonant X-ray inelastic scattering studies on solid materials, liquids and gases at diffraction-limited storage rings.

    PubMed

    Schmitt, Thorsten; de Groot, Frank M F; Rubensson, Jan Erik

    2014-09-01

    The spectroscopic technique of resonant inelastic X-ray scattering (RIXS) will particularly profit from immensely improved brilliance of diffraction-limited storage rings (DLSRs). In RIXS one measures the intensities of excitations as a function of energy and momentum transfer. DLSRs will allow for pushing the achievable energy resolution, signal intensity and the sampled spot size to new limits. With RIXS one nowadays probes a broad range of electronic systems reaching from simple molecules to complex materials displaying phenomena like peculiar magnetism, two-dimensional electron gases, superconductivity, photovoltaic energy conversion and heterogeneous catalysis. In this article the types of improved RIXS studies that will become possible with X-ray beams from DLSRs are envisioned.

  12. Novel radio-frequency gun structures for ultrafast relativistic electron diffraction.

    PubMed

    Musumeci, P; Faillace, L; Fukasawa, A; Moody, J T; O'Shea, B; Rosenzweig, J B; Scoby, C M

    2009-08-01

    Radio-frequency (RF) photoinjector-based relativistic ultrafast electron diffraction (UED) is a promising new technique that has the potential to probe structural changes at the atomic scale with sub-100 fs temporal resolution in a single shot. We analyze the limitations on the temporal and spatial resolution of this technique considering the operating parameters of a standard 1.6 cell RF gun (which is the RF photoinjector used for the first experimental tests of relativistic UED at Stanford Linear Accelerator Center; University of California, Los Angeles; Brookhaven National Laboratory), and study the possibility of employing novel RF structures to circumvent some of these limits.

  13. Coherent X-ray beam metrology using 2D high-resolution Fresnel-diffraction analysis.

    PubMed

    Ruiz-Lopez, M; Faenov, A; Pikuz, T; Ozaki, N; Mitrofanov, A; Albertazzi, B; Hartley, N; Matsuoka, T; Ochante, Y; Tange, Y; Yabuuchi, T; Habara, T; Tanaka, K A; Inubushi, Y; Yabashi, M; Nishikino, M; Kawachi, T; Pikuz, S; Ishikawa, T; Kodama, R; Bleiner, D

    2017-01-01

    Direct metrology of coherent short-wavelength beamlines is important for obtaining operational beam characteristics at the experimental site. However, since beam-time limitation imposes fast metrology procedures, a multi-parametric metrology from as low as a single shot is desirable. Here a two-dimensional (2D) procedure based on high-resolution Fresnel diffraction analysis is discussed and applied, which allowed an efficient and detailed beamline characterization at the SACLA XFEL. So far, the potential of Fresnel diffraction for beamline metrology has not been fully exploited because its high-frequency fringes could be only partly resolved with ordinary pixel-limited detectors. Using the high-spatial-frequency imaging capability of an irradiated LiF crystal, 2D information of the coherence degree, beam divergence and beam quality factor M 2 were retrieved from simple diffraction patterns. The developed beam metrology was validated with a laboratory reference laser, and then successfully applied at a beamline facility, in agreement with the source specifications.

  14. Accessing High Spatial Resolution in Astronomy Using Interference Methods

    NASA Astrophysics Data System (ADS)

    Carbonel, Cyril; Grasset, Sébastien; Maysonnave, Jean

    2018-04-01

    In astronomy, methods such as direct imaging or interferometry-based techniques (Michelson stellar interferometry for example) are used for observations. A particular advantage of interferometry is that it permits greater spatial resolution compared to direct imaging with a single telescope, which is limited by diffraction owing to the aperture of the instrument as shown by Rueckner et al. in a lecture demonstration. The focus of this paper, addressed to teachers and/or students in high schools and universities, is to easily underline both an application of interferometry in astronomy and stress its interest for resolution. To this end very simple optical experiments are presented to explain all the concepts. We show how an interference pattern resulting from the combined signals of two telescopes allows us to measure the distance between two stars with a resolution beyond the diffraction limit. Finally this work emphasizes the breathtaking resolution obtained in state-of-the-art instruments such as the VLTi (Very Large Telescope interferometer).

  15. Experimental observation of sub-Rayleigh quantum imaging with a two-photon entangled source

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xu, De-Qin; School of Science, Tianjin University of Technology and Education, Tianjin 300222; Song, Xin-Bing

    It has been theoretically predicted that N-photon quantum imaging can realize either an N-fold resolution improvement (Heisenberg-like scaling) or a √(N)-fold resolution improvement (standard quantum limit) beyond the Rayleigh diffraction bound, over classical imaging. Here, we report the experimental study on spatial sub-Rayleigh quantum imaging using a two-photon entangled source. Two experimental schemes are proposed and performed. In a Fraunhofer diffraction scheme with a lens, two-photon Airy disk pattern is observed with subwavelength diffraction property. In a lens imaging apparatus, however, two-photon sub-Rayleigh imaging for an object is realized with super-resolution property. The experimental results agree with the theoretical predictionmore » in the two-photon quantum imaging regime.« less

  16. Large scale superres 3D imaging: light-sheet single-molecule localization microscopy (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Lu, Chieh Han; Chen, Peilin; Chen, Bi-Chang

    2017-02-01

    Optical imaging techniques provide much important information in understanding life science especially cellular structure and morphology because "seeing is believing". However, the resolution of optical imaging is limited by the diffraction limit, which is discovered by Ernst Abbe, i.e. λ/2(NA) (NA is the numerical aperture of the objective lens). Fluorescence super-resolution microscopic techniques such as Stimulated emission depletion microscopy (STED), Photoactivated localization microscopy (PALM), and Stochastic optical reconstruction microscopy (STORM) are invented to have the capability of seeing biological entities down to molecular level that are smaller than the diffraction limit (around 200-nm in lateral resolution). These techniques do not physically violate the Abbe limit of resolution but exploit the photoluminescence properties and labelling specificity of fluorescence molecules to achieve super-resolution imaging. However, these super-resolution techniques limit most of their applications to the 2D imaging of fixed or dead samples due to the high laser power needed or slow speed for the localization process. Extended from 2D imaging, light sheet microscopy has been proven to have a lot of applications on 3D imaging at much better spatiotemporal resolutions due to its intrinsic optical sectioning and high imaging speed. Herein, we combine the advantage of localization microscopy and light-sheet microscopy to have super-resolved cellular imaging in 3D across large field of view. With high-density labeled spontaneous blinking fluorophore and wide-field detection of light-sheet microscopy, these allow us to construct 3D super-resolution multi-cellular imaging at high speed ( minutes) by light-sheet single-molecule localization microscopy.

  17. STED super-resolution microscopy of clinical paraffin-embedded human rectal cancer tissue.

    PubMed

    Ilgen, Peter; Stoldt, Stefan; Conradi, Lena-Christin; Wurm, Christian Andreas; Rüschoff, Josef; Ghadimi, B Michael; Liersch, Torsten; Jakobs, Stefan

    2014-01-01

    Formalin fixed and paraffin-embedded human tissue resected during cancer surgery is indispensable for diagnostic and therapeutic purposes and represents a vast and largely unexploited resource for research. Optical microscopy of such specimen is curtailed by the diffraction-limited resolution of conventional optical microscopy. To overcome this limitation, we used STED super-resolution microscopy enabling optical resolution well below the diffraction barrier. We visualized nanoscale protein distributions in sections of well-annotated paraffin-embedded human rectal cancer tissue stored in a clinical repository. Using antisera against several mitochondrial proteins, STED microscopy revealed distinct sub-mitochondrial protein distributions, suggesting a high level of structural preservation. Analysis of human tissues stored for up to 17 years demonstrated that these samples were still amenable for super-resolution microscopy. STED microscopy of sections of HER2 positive rectal adenocarcinoma revealed details in the surface and intracellular HER2 distribution that were blurred in the corresponding conventional images, demonstrating the potential of super-resolution microscopy to explore the thus far largely untapped nanoscale regime in tissues stored in biorepositories.

  18. STED Super-Resolution Microscopy of Clinical Paraffin-Embedded Human Rectal Cancer Tissue

    PubMed Central

    Wurm, Christian Andreas; Rüschoff, Josef; Ghadimi, B. Michael; Liersch, Torsten; Jakobs, Stefan

    2014-01-01

    Formalin fixed and paraffin-embedded human tissue resected during cancer surgery is indispensable for diagnostic and therapeutic purposes and represents a vast and largely unexploited resource for research. Optical microscopy of such specimen is curtailed by the diffraction-limited resolution of conventional optical microscopy. To overcome this limitation, we used STED super-resolution microscopy enabling optical resolution well below the diffraction barrier. We visualized nanoscale protein distributions in sections of well-annotated paraffin-embedded human rectal cancer tissue stored in a clinical repository. Using antisera against several mitochondrial proteins, STED microscopy revealed distinct sub-mitochondrial protein distributions, suggesting a high level of structural preservation. Analysis of human tissues stored for up to 17 years demonstrated that these samples were still amenable for super-resolution microscopy. STED microscopy of sections of HER2 positive rectal adenocarcinoma revealed details in the surface and intracellular HER2 distribution that were blurred in the corresponding conventional images, demonstrating the potential of super-resolution microscopy to explore the thus far largely untapped nanoscale regime in tissues stored in biorepositories. PMID:25025184

  19. Modeling the depth-sectioning effect in reflection-mode dynamic speckle-field interferometric microscopy

    PubMed Central

    Zhou, Renjie; Jin, Di; Hosseini, Poorya; Singh, Vijay Raj; Kim, Yang-hyo; Kuang, Cuifang; Dasari, Ramachandra R.; Yaqoob, Zahid; So, Peter T. C.

    2017-01-01

    Unlike most optical coherence microscopy (OCM) systems, dynamic speckle-field interferometric microscopy (DSIM) achieves depth sectioning through the spatial-coherence gating effect. Under high numerical aperture (NA) speckle-field illumination, our previous experiments have demonstrated less than 1 μm depth resolution in reflection-mode DSIM, while doubling the diffraction limited resolution as under structured illumination. However, there has not been a physical model to rigorously describe the speckle imaging process, in particular explaining the sectioning effect under high illumination and imaging NA settings in DSIM. In this paper, we develop such a model based on the diffraction tomography theory and the speckle statistics. Using this model, we calculate the system response function, which is used to further obtain the depth resolution limit in reflection-mode DSIM. Theoretically calculated depth resolution limit is in an excellent agreement with experiment results. We envision that our physical model will not only help in understanding the imaging process in DSIM, but also enable better designing such systems for depth-resolved measurements in biological cells and tissues. PMID:28085800

  20. Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells

    PubMed Central

    Rodriguez, Jose A.; Xu, Rui; Chen, Chien-Chun; Huang, Zhifeng; Jiang, Huaidong; Chen, Allan L.; Raines, Kevin S.; Pryor Jr, Alan; Nam, Daewoong; Wiegart, Lutz; Song, Changyong; Madsen, Anders; Chushkin, Yuriy; Zontone, Federico; Bradley, Peter J.; Miao, Jianwei

    2015-01-01

    A structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 keV X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and the three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. It is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres. PMID:26306199

  1. Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells

    DOE PAGES

    Rodriguez, Jose A.; Xu, Rui; Chen, Chien -Chun; ...

    2015-09-01

    Here, a structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 Kev X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and themore » three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. Finally, it is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres.« less

  2. Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells.

    PubMed

    Rodriguez, Jose A; Xu, Rui; Chen, Chien-Chun; Huang, Zhifeng; Jiang, Huaidong; Chen, Allan L; Raines, Kevin S; Pryor, Alan; Nam, Daewoong; Wiegart, Lutz; Song, Changyong; Madsen, Anders; Chushkin, Yuriy; Zontone, Federico; Bradley, Peter J; Miao, Jianwei

    2015-09-01

    A structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 keV X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and the three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. It is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres.

  3. From Airy to Abbe: quantifying the effects of wide-angle focusing for scalar spherical waves

    NASA Astrophysics Data System (ADS)

    Calm, Yitzi M.; Merlo, Juan M.; Burns, Michael J.; Naughton, Michael J.

    2017-10-01

    Recent advances in optical microscopy have enabled imaging with spatial resolution beyond the diffraction limit. This limit is sometimes taken as one of several different criteria according to different conventions, including Rayleigh’s 0.61λ /NA, Abbe’s 0.5λ /NA, and Sparrow’s 0.47λ /NA. In this paper, we perform a parametric study, numerically integrating the scalar Kirchhoff diffraction integrals, and we propose new functional forms for the resolution limits derived from scalar focusing. The new expressions remain accurate under wide angle focusing, up to 90^\\circ . Our results could materially impact the design of high intensity focused ultrasound systems, and can be used as a qualitative guideline for the design of a particular type of planar optical element: the flat lens metasurface.

  4. Prospects of high-resolution resonant X-ray inelastic scattering studies on solid materials, liquids and gases at diffraction-limited storage rings

    PubMed Central

    Schmitt, Thorsten; de Groot, Frank M. F.; Rubensson, Jan-Erik

    2014-01-01

    The spectroscopic technique of resonant inelastic X-ray scattering (RIXS) will particularly profit from immensely improved brilliance of diffraction-limited storage rings (DLSRs). In RIXS one measures the intensities of excitations as a function of energy and momentum transfer. DLSRs will allow for pushing the achievable energy resolution, signal intensity and the sampled spot size to new limits. With RIXS one nowadays probes a broad range of electronic systems reaching from simple molecules to complex materials displaying phenomena like peculiar magnetism, two-dimensional electron gases, superconductivity, photovoltaic energy conversion and heterogeneous catalysis. In this article the types of improved RIXS studies that will become possible with X-ray beams from DLSRs are envisioned. PMID:25177995

  5. Coherent total internal reflection dark-field microscopy: label-free imaging beyond the diffraction limit.

    PubMed

    von Olshausen, Philipp; Rohrbach, Alexander

    2013-10-15

    Coherent imaging is barely applicable in life-science microscopy due to multiple interference artifacts. Here, we show how these interferences can be used to improve image resolution and contrast. We present a dark-field microscopy technique with evanescent illumination via total internal reflection that delivers high-contrast images of coherently scattering samples. By incoherent averaging of multiple coherent images illuminated from different directions we can resolve image structures that remain unresolved by conventional (incoherent) fluorescence microscopy. We provide images of 190 nm beads revealing resolution beyond the diffraction limit and slightly increased object distances. An analytical model is introduced that accounts for the observed effects and which is confirmed by numerical simulations. Our approach may be a route to fast, label-free, super-resolution imaging in live-cell microscopy.

  6. Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT

    PubMed Central

    Hoyer, Patrick; de Medeiros, Gustavo; Balázs, Bálint; Norlin, Nils; Besir, Christina; Hanne, Janina; Kräusslich, Hans-Georg; Engelhardt, Johann; Sahl, Steffen J.; Hell, Stefan W.; Hufnagel, Lars

    2016-01-01

    We present a plane-scanning RESOLFT [reversible saturable/switchable optical (fluorescence) transitions] light-sheet (LS) nanoscope, which fundamentally overcomes the diffraction barrier in the axial direction via confinement of the fluorescent molecular state to a sheet of subdiffraction thickness around the focal plane. To this end, reversibly switchable fluorophores located right above and below the focal plane are transferred to a nonfluorescent state at each scanning step. LS-RESOLFT nanoscopy offers wide-field 3D imaging of living biological specimens with low light dose and axial resolution far beyond the diffraction barrier. We demonstrate optical sections that are thinner by 5–12-fold compared with their conventional diffraction-limited LS analogs. PMID:26984498

  7. Modeling super-resolution SERS using a T-matrix method to elucidate molecule-nanoparticle coupling and the origins of localization errors

    NASA Astrophysics Data System (ADS)

    Heaps, Charles W.; Schatz, George C.

    2017-06-01

    A computational method to model diffraction-limited images from super-resolution surface-enhanced Raman scattering microscopy is introduced. Despite significant experimental progress in plasmon-based super-resolution imaging, theoretical predictions of the diffraction limited images remain a challenge. The method is used to calculate localization errors and image intensities for a single spherical gold nanoparticle-molecule system. The light scattering is calculated using a modification of generalized Mie (T-matrix) theory with a point dipole source and diffraction limited images are calculated using vectorial diffraction theory. The calculation produces the multipole expansion for each emitter and the coherent superposition of all fields. Imaging the constituent fields in addition to the total field provides new insight into the strong coupling between the molecule and the nanoparticle. Regardless of whether the molecular dipole moment is oriented parallel or perpendicular to the nanoparticle surface, the anisotropic excitation distorts the center of the nanoparticle as measured by the point spread function by approximately fifty percent of the particle radius toward to the molecule. Inspection of the nanoparticle multipoles reveals that distortion arises from a weak quadrupole resonance interfering with the dipole field in the nanoparticle. When the nanoparticle-molecule fields are in-phase, the distorted nanoparticle field dominates the observed image. When out-of-phase, the nanoparticle and molecule are of comparable intensity and interference between the two emitters dominates the observed image. The method is also applied to different wavelengths and particle radii. At off-resonant wavelengths, the method predicts images closer to the molecule not because of relative intensities but because of greater distortion in the nanoparticle. The method is a promising approach to improving the understanding of plasmon-enhanced super-resolution experiments.

  8. Near-field limitations of Fresnel-regime coherent diffraction imaging

    DOE PAGES

    Pound, Benjamin A.; Barber, John L.; Nguyen, Kimberly; ...

    2017-08-04

    Coherent diffraction imaging (CDI) is a rapidly developing form of imaging that offers the potential of wavelength-limited resolution without image-forming lenses. In CDI, the intensity of the diffraction pattern is measured directly by the detector, and various iterative phase retrieval algorithms are used to “invert” the diffraction pattern and reconstruct a high-resolution image of the sample. But, there are certain requirements in CDI that must be met to reconstruct the object. Although most experiments are conducted in the “far-field”—or Fraunhofer—regime where the requirements are not as stringent, some experiments must be conducted in the “near field” where Fresnel diffraction mustmore » be considered. According to the derivation of Fresnel diffraction, successful reconstructions can only be obtained when the small-angle number, a derived quantity, is much less than one. We show, however, that it is not actually necessary to fulfill the small-angle condition. The Fresnel kernel well approximates the exact kernel in regions where the phase oscillates slowly, and in regions of fast oscillations, indicated by large A n , the error between kernels should be negligible due to stationary-phase arguments. Finally we verify, by experiment, this conclusion with a helium neon laser setup and show that it should hold at x-ray wavelengths as well.« less

  9. Near-field limitations of Fresnel-regime coherent diffraction imaging

    NASA Astrophysics Data System (ADS)

    Pound, Benjamin A.; Barber, John L.; Nguyen, Kimberly; Tyson, Matthew C.; Sandberg, Richard L.

    2017-08-01

    Coherent diffraction imaging (CDI) is a rapidly developing form of imaging that offers the potential of wavelength-limited resolution without image-forming lenses. In CDI, the intensity of the diffraction pattern is measured directly by the detector, and various iterative phase retrieval algorithms are used to "invert" the diffraction pattern and reconstruct a high-resolution image of the sample. However, there are certain requirements in CDI that must be met to reconstruct the object. Although most experiments are conducted in the "far-field"—or Fraunhofer—regime where the requirements are not as stringent, some experiments must be conducted in the "near field" where Fresnel diffraction must be considered. According to the derivation of Fresnel diffraction, successful reconstructions can only be obtained when the small-angle number, a derived quantity, is much less than one. We show, however, that it is not actually necessary to fulfill the small-angle condition. The Fresnel kernel well approximates the exact kernel in regions where the phase oscillates slowly, and in regions of fast oscillations, indicated by large A n , the error between kernels should be negligible due to stationary-phase arguments. We experimentally verify this conclusion with a helium neon laser setup and show that it should hold at x-ray wavelengths as well.

  10. Sub-diffraction limit laser ablation via multiple exposures using a digital micromirror device.

    PubMed

    Heath, Daniel J; Grant-Jacob, James A; Feinaeugle, Matthias; Mills, Ben; Eason, Robert W

    2017-08-01

    We present the use of digital micromirror devices as variable illumination masks for pitch-splitting multiple exposures to laser machine the surfaces of materials. Ultrafast laser pulses of length 150 fs and 800 nm central wavelength were used for the sequential machining of contiguous patterns on the surface of samples in order to build up complex structures with sub-diffraction limit features. Machined patterns of tens to hundreds of micrometers in lateral dimensions with feature separations as low as 270 nm were produced in electroless nickel on an optical setup diffraction limited to 727 nm, showing a reduction factor below the Abbe diffraction limit of ∼2.7×. This was compared to similar patterns in a photoresist optimized for two-photon absorption, which showed a reduction factor of only 2×, demonstrating that multiple exposures via ablation can produce a greater resolution enhancement than via two-photon polymerization.

  11. Protein crystal structure obtained at 2.9 Å resolution from injecting bacterial cells into an X-ray free-electron laser beam

    PubMed Central

    Sawaya, Michael R.; Cascio, Duilio; Gingery, Mari; Rodriguez, Jose; Goldschmidt, Lukasz; Colletier, Jacques-Philippe; Messerschmidt, Marc M.; Boutet, Sébastien; Koglin, Jason E.; Williams, Garth J.; Brewster, Aaron S.; Nass, Karol; Hattne, Johan; Botha, Sabine; Doak, R. Bruce; Shoeman, Robert L.; DePonte, Daniel P.; Park, Hyun-Woo; Federici, Brian A.; Sauter, Nicholas K.; Schlichting, Ilme; Eisenberg, David S.

    2014-01-01

    It has long been known that toxins produced by Bacillus thuringiensis (Bt) are stored in the bacterial cells in crystalline form. Here we describe the structure determination of the Cry3A toxin found naturally crystallized within Bt cells. When whole Bt cells were streamed into an X-ray free-electron laser beam we found that scattering from other cell components did not obscure diffraction from the crystals. The resolution limits of the best diffraction images collected from cells were the same as from isolated crystals. The integrity of the cells at the moment of diffraction is unclear; however, given the short time (∼5 µs) between exiting the injector to intersecting with the X-ray beam, our result is a 2.9-Å-resolution structure of a crystalline protein as it exists in a living cell. The study suggests that authentic in vivo diffraction studies can produce atomic-level structural information. PMID:25136092

  12. Planar super-oscillatory lens for sub-diffraction optical needles at violet wavelengths

    PubMed Central

    Yuan, Guanghui; Rogers, Edward T. F.; Roy, Tapashree; Adamo, Giorgio; Shen, Zexiang; Zheludev, Nikolay I.

    2014-01-01

    Planar optical lenses are fundamental elements of miniaturized photonic devices. However, conventional planar optical lenses are constrained by the diffraction limit in the optical far-field due to the band-limited wavevectors supported by free-space and loss of high-spatial-frequency evanescent components. As inspired by Einstein's radiation ‘needle stick', electromagnetic energy can be delivered into an arbitrarily small solid angle. Such sub-diffraction optical needles have been numerically investigated using diffractive optical elements (DOEs) together with specially polarized optical beams, but experimental demonstration is extremely difficult due to the bulky size of DOEs and the required alignment precision. Planar super-oscillatory lenses (SOLs) were proposed to overcome these constraints and demonstrated that sub-diffraction focal spots can actually be formed without any evanescent waves, making far-field, label-free super-resolution imaging possible. Here we extend the super-oscillation concept into the vectorial-field regime to work with circularly polarized light, and experimentally demonstrate, for the first time, a circularly polarized optical needle with sub-diffraction transverse spot size (0.45λ) and axial long depth of focus (DOF) of 15λ using a planar SOL at a violet wavelength of 405 nm. This sub-diffraction circularly polarized optical needle has potential applications in circular dichroism spectroscopy, super-resolution imaging, high-density optical storage, heat-assisted magnetic recording, nano-manufacturing and nano-metrology. PMID:25208611

  13. Rayleigh-wave diffractions due to a void in the layered half space

    USGS Publications Warehouse

    Xia, J.; Xu, Y.; Miller, R.D.; Nyquist, Jonathan E.

    2006-01-01

    Void detection is challenging due to the complexity of near-surface materials and the limited resolution of geophysical methods. Although multichannel, high-frequency, surface-wave techniques can provide reliable shear (S)-wave velocities in different geological settings, they are not suitable for detecting voids directly based on anomalies of the S-wave velocity because of limitations on the resolution of S-wave velocity profiles inverted from surface-wave phase velocities. Xia et al. (2006a) derived a Rayleigh-wave diffraction traveltime equation due to a void in the homogeneous half space. Encouraging results of directly detecting a void from Rayleigh-wave diffractions were presented (Xia et al., 2006a). In this paper we used four two-dimensional square voids in the layered half space to demonstrate the feasibility of detecting a void with Rayleigh-wave diffractions. Rayleigh-wave diffractions were recognizable for all these models after removing direct surface waves by F-K filtering. We evaluate the feasibility of applying the Rayleigh-wave diffraction traveltime equation to a void in the layered earth model. The phase velocity of diffracted Rayleigh waves is predominately determined by surrounding materials of a void. The modeling results demonstrate that the Rayleigh-wave diffraction traveltime equation due to a void in the homogeneous half space can be applied to the case of a void in the layered half space. In practice, only two diffraction times are necessary to define the depth to the top of a void and the average velocity of diffracted Rayleigh waves. ?? 2005 Society of Exploration Geophysicists.

  14. Coherent diffractive imaging of time-evolving samples with improved temporal resolution

    DOE PAGES

    Ulvestad, A.; Tripathi, A.; Hruszkewycz, S. O.; ...

    2016-05-19

    Bragg coherent x-ray diffractive imaging is a powerful technique for investigating dynamic nanoscale processes in nanoparticles immersed in reactive, realistic environments. Its temporal resolution is limited, however, by the oversampling requirements of three-dimensional phase retrieval. Here, we show that incorporating the entire measurement time series, which is typically a continuous physical process, into phase retrieval allows the oversampling requirement at each time step to be reduced, leading to a subsequent improvement in the temporal resolution by a factor of 2-20 times. The increased time resolution will allow imaging of faster dynamics and of radiation-dose-sensitive samples. Furthermore, this approach, which wemore » call "chrono CDI," may find use in improving the time resolution in other imaging techniques.« less

  15. Sub-diffraction limit resolution in microscopy

    NASA Technical Reports Server (NTRS)

    Cheng, Ming (Inventor); Chen, Weinong (Inventor)

    2007-01-01

    A method and apparatus for visualizing sub-micron size particles employs a polarizing microscope wherein a focused beam of polarized light is projected onto a target, and a portion of the illuminating light is blocked from reaching the specimen, whereby to produce a shadow region, and projecting diffracted light from the target onto the shadow region.

  16. Scanning superlens microscopy for non-invasive large field-of-view visible light nanoscale imaging

    NASA Astrophysics Data System (ADS)

    Wang, Feifei; Liu, Lianqing; Yu, Haibo; Wen, Yangdong; Yu, Peng; Liu, Zhu; Wang, Yuechao; Li, Wen Jung

    2016-12-01

    Nanoscale correlation of structural information acquisition with specific-molecule identification provides new insight for studying rare subcellular events. To achieve this correlation, scanning electron microscopy has been combined with super-resolution fluorescent microscopy, despite its destructivity when acquiring biological structure information. Here we propose time-efficient non-invasive microsphere-based scanning superlens microscopy that enables the large-area observation of live-cell morphology or sub-membrane structures with sub-diffraction-limited resolution and is demonstrated by observing biological and non-biological objects. This microscopy operates in both non-invasive and contact modes with ~200 times the acquisition efficiency of atomic force microscopy, which is achieved by replacing the point of an atomic force microscope tip with an imaging area of microspheres and stitching the areas recorded during scanning, enabling sub-diffraction-limited resolution. Our method marks a possible path to non-invasive cell imaging and simultaneous tracking of specific molecules with nanoscale resolution, facilitating the study of subcellular events over a total cell period.

  17. High resolution telescope including an array of elemental telescopes aligned along a common axis and supported on a space frame with a pivot at its geometric center

    DOEpatents

    Norbert, M.A.; Yale, O.

    1992-04-28

    A large effective-aperture, low-cost optical telescope with diffraction-limited resolution enables ground-based observation of near-earth space objects. The telescope has a non-redundant, thinned-aperture array in a center-mount, single-structure space frame. It employes speckle interferometric imaging to achieve diffraction-limited resolution. The signal-to-noise ratio problem is mitigated by moving the wavelength of operation to the near-IR, and the image is sensed by a Silicon CCD. The steerable, single-structure array presents a constant pupil. The center-mount, radar-like mount enables low-earth orbit space objects to be tracked as well as increases stiffness of the space frame. In the preferred embodiment, the array has elemental telescopes with subaperture of 2.1 m in a circle-of-nine configuration. The telescope array has an effective aperture of 12 m which provides a diffraction-limited resolution of 0.02 arc seconds. Pathlength matching of the telescope array is maintained by a electro-optical system employing laser metrology. Speckle imaging relaxes pathlength matching tolerance by one order of magnitude as compared to phased arrays. Many features of the telescope contribute to substantial reduction in costs. These include eliminating the conventional protective dome and reducing on-site construction activities. The cost of the telescope scales with the first power of the aperture rather than its third power as in conventional telescopes. 15 figs.

  18. High resolution telescope including an array of elemental telescopes aligned along a common axis and supported on a space frame with a pivot at its geometric center

    DOEpatents

    Norbert, Massie A.; Yale, Oster

    1992-01-01

    A large effective-aperture, low-cost optical telescope with diffraction-limited resolution enables ground-based observation of near-earth space objects. The telescope has a non-redundant, thinned-aperture array in a center-mount, single-structure space frame. It employes speckle interferometric imaging to achieve diffraction-limited resolution. The signal-to-noise ratio problem is mitigated by moving the wavelength of operation to the near-IR, and the image is sensed by a Silicon CCD. The steerable, single-structure array presents a constant pupil. The center-mount, radar-like mount enables low-earth orbit space objects to be tracked as well as increases stiffness of the space frame. In the preferred embodiment, the array has elemental telescopes with subaperture of 2.1 m in a circle-of-nine configuration. The telescope array has an effective aperture of 12 m which provides a diffraction-limited resolution of 0.02 arc seconds. Pathlength matching of the telescope array is maintained by a electro-optical system employing laser metrology. Speckle imaging relaxes pathlength matching tolerance by one order of magnitude as compared to phased arrays. Many features of the telescope contribute to substantial reduction in costs. These include eliminating the conventional protective dome and reducing on-site construction activities. The cost of the telescope scales with the first power of the aperture rather than its third power as in conventional telescopes.

  19. Sub-micron materials characterization using near-field optics

    NASA Astrophysics Data System (ADS)

    Blodgett, David Wesley

    1998-12-01

    High-resolution sub-surface materials characterization and inspection are critical in the microelectronics and thin films industries. To this end, a technique is described that couples the bulk property measurement capabilities of high-frequency ultrasound with the high-resolution surface imaging capabilities of the near-field optical microscope. Sensing bulk microstructure variations in the material, such as grain boundaries, requires a detection footprint smaller than the variation itself. The near-field optical microscope, with the ability to exceed the diffraction limit in optical resolution, meets this requirement. Two apertureless near-field optical microscopes, on-axis and off-axis illumination, have been designed and built. Near-field and far-field approach curves for both microscopes are presented. The sensitivity of the near-field approach curve was 8.3 muV/nm. Resolution studies for the near-field microscope indicate optical resolutions on the order of 50 nm, which exceeds the diffraction limit. The near-field microscope has been adapted to detect both contact-transducer-generated and laser-generated ultrasound. The successful detection of high-frequency ultrasound with the near-field optical microscope demonstrates the potential of this technique.

  20. Design and analysis of a fast, two-mirror soft-x-ray microscope

    NASA Technical Reports Server (NTRS)

    Shealy, D. L.; Wang, C.; Jiang, W.; Jin, L.; Hoover, R. B.

    1992-01-01

    During the past several years, a number of investigators have addressed the design, analysis, fabrication, and testing of spherical Schwarzschild microscopes for soft-x-ray applications using multilayer coatings. Some of these systems have demonstrated diffraction limited resolution for small numerical apertures. Rigorously aplanatic, two-aspherical mirror Head microscopes can provide near diffraction limited resolution for very large numerical apertures. The relationships between the numerical aperture, mirror radii and diameters, magnifications, and total system length for Schwarzschild microscope configurations are summarized. Also, an analysis of the characteristics of the Head-Schwarzschild surfaces will be reported. The numerical surface data predicted by the Head equations were fit by a variety of functions and analyzed by conventional optical design codes. Efforts have been made to determine whether current optical substrate and multilayer coating technologies will permit construction of a very fast Head microscope which can provide resolution approaching that of the wavelength of the incident radiation.

  1. Multi-color localization microscopy of fixed cells as a promising tool to study organization of bacterial cytoskeleton

    NASA Astrophysics Data System (ADS)

    Vedyaykin, A. D.; Gorbunov, V. V.; Sabantsev, A. V.; Polinovskaya, V. S.; Vishnyakov, I. E.; Melnikov, A. S.; Serdobintsev, P. Yu; Khodorkovskii, M. A.

    2015-11-01

    Localization microscopy allows visualization of biological structures with resolution well below the diffraction limit. Localization microscopy was used to study FtsZ organization in Escherichia coli previously in combination with fluorescent protein labeling, but the fact that fluorescent chimeric protein was unable to rescue temperature-sensitive ftsZ mutants suggests that obtained images may not represent native FtsZ structures faithfully. Indirect immunolabeling of FtsZ not only overcomes this problem, but also allows the use of the powerful visualization methods arsenal available for different structures in fixed cells. In this work we simultaneously obtained super-resolution images of FtsZ structures and diffraction-limited or super-resolution images of DNA and cell surface in E. coli, which allows for the study of the spatial arrangement of FtsZ structures with respect to the nucleoid positions and septum formation.

  2. Diffraction-limited Mid-infrared Integral Field Spectroscopy of Io's Volcanic Activity with ALES on the Large Binocular Telescope

    NASA Astrophysics Data System (ADS)

    Skrutskie, Michael F.; de Kleer, Katherine R.; Stone, Jordan; Conrad, Al; Davies, Ashley; de Pater, Imke; Leisenring, Jarron; Hinz, Philip; Skemer, Andrew; Veillet, Christian; Woodward, Charles E.; Ertel, Steve; Spalding, Eckhart

    2017-10-01

    The Arizona Lenslet for Exoplanet Spectroscopy (ALES) is an enhancement to the Large Binocular Telescope's mid-infrared imager, LMIRcam, that permits low-resolution (R~20) spectroscopy between 2.8 and 4.2 μm of every diffraction-limited resolution element in a 2.5"x2.5" field-of-view on a 2048x2048 HAWAII-2RG 5.2 μm-cutoff array. The 1" disk of Io, dotted with powerful self-luminous volcanic eruptions, provides an ideal target for ALES, where the single 8.4-meter aperture diffraction-limited scale for Io at opposition ranges from 240 kilometers (80 milliarcseconds) at 2.8 μm to 360 kilometers (120 milliarcseconds) at 4.2 μm. ALES provides the capability to assess the color temperature of each volcanic thermal emission site as well as map broadband absorbers such as SO2 frost. A monitoring campaign in the Spring 2017 semester provided two global snapshots of Io's volcanic activity with ALES as well as characterization of a new brightening episode at Loki Patera over four epochs between January and May 2017.

  3. Pushing the plasmonic imaging nanolithography to nano-manufacturing

    NASA Astrophysics Data System (ADS)

    Gao, Ping; Li, Xiong; Zhao, Zeyu; Ma, Xiaoliang; Pu, Mingbo; Wang, Changtao; Luo, Xiangang

    2017-12-01

    Suffering from the so-called diffraction limit, the minimum resolution of conventional photolithography is limited to λ / 2 or λ / 4, where λ is the incident wavelength. The physical mechanism of this limit lies at the fact that the evanescent waves that carry subwavelength information of the object decay exponentially in a medium, and cannot reach the image plane. Surface plasmons (SPs) are non-radiative electromagnetic waves that propagate along the interface between metal and dielectric, which exhibits unique sub-diffraction optical characteristics. In recent years, benefiting from SPs' features, researchers have proposed a variety of plasmonic lithography methods in the manner of interference, imaging and direct writing, and have demonstrated that sub-diffraction resolution could be achieved by theoretical simulations or experiments. Among the various plasmonic lithography modes, plasmonic imaging lithography seems to be of particular importance for applications due to its compatibility with conventional lithography. Recent results show that the half pitch of nanograting can be shrinked down to 22 nm and even 16 nm. This paper will give an overview of research progress, representative achievements of plasmonic imaging lithography, the remained problems and outlook of further developments.

  4. Grain rotation and lattice deformation during photoinduced chemical reactions revealed by in situ X-ray nanodiffraction.

    PubMed

    Huang, Zhifeng; Bartels, Matthias; Xu, Rui; Osterhoff, Markus; Kalbfleisch, Sebastian; Sprung, Michael; Suzuki, Akihiro; Takahashi, Yukio; Blanton, Thomas N; Salditt, Tim; Miao, Jianwei

    2015-07-01

    In situ X-ray diffraction (XRD) and transmission electron microscopy (TEM) have been used to investigate many physical science phenomena, ranging from phase transitions, chemical reactions and crystal growth to grain boundary dynamics. A major limitation of in situ XRD and TEM is a compromise that has to be made between spatial and temporal resolution. Here, we report the development of in situ X-ray nanodiffraction to measure high-resolution diffraction patterns from single grains with up to 5 ms temporal resolution. We observed, for the first time, grain rotation and lattice deformation in chemical reactions induced by X-ray photons: Br(-) + hv → Br + e(-) and e(-) + Ag(+) → Ag(0). The grain rotation and lattice deformation associated with the chemical reactions were quantified to be as fast as 3.25 rad s(-1) and as large as 0.5 Å, respectively. The ability to measure high-resolution diffraction patterns from individual grains with a temporal resolution of several milliseconds is expected to find broad applications in materials science, physics, chemistry and nanoscience.

  5. Design and implementation of an optimal laser pulse front tilting scheme for ultrafast electron diffraction in reflection geometry with high temporal resolution.

    PubMed

    Pennacchio, Francesco; Vanacore, Giovanni M; Mancini, Giulia F; Oppermann, Malte; Jayaraman, Rajeswari; Musumeci, Pietro; Baum, Peter; Carbone, Fabrizio

    2017-07-01

    Ultrafast electron diffraction is a powerful technique to investigate out-of-equilibrium atomic dynamics in solids with high temporal resolution. When diffraction is performed in reflection geometry, the main limitation is the mismatch in group velocity between the overlapping pump light and the electron probe pulses, which affects the overall temporal resolution of the experiment. A solution already available in the literature involved pulse front tilt of the pump beam at the sample, providing a sub-picosecond time resolution. However, in the reported optical scheme, the tilted pulse is characterized by a temporal chirp of about 1 ps at 1 mm away from the centre of the beam, which limits the investigation of surface dynamics in large crystals. In this paper, we propose an optimal tilting scheme designed for a radio-frequency-compressed ultrafast electron diffraction setup working in reflection geometry with 30 keV electron pulses containing up to 10 5 electrons/pulse. To characterize our scheme, we performed optical cross-correlation measurements, obtaining an average temporal width of the tilted pulse lower than 250 fs. The calibration of the electron-laser temporal overlap was obtained by monitoring the spatial profile of the electron beam when interacting with the plasma optically induced at the apex of a copper needle (plasma lensing effect). Finally, we report the first time-resolved results obtained on graphite, where the electron-phonon coupling dynamics is observed, showing an overall temporal resolution in the sub-500 fs regime. The successful implementation of this configuration opens the way to directly probe structural dynamics of low-dimensional systems in the sub-picosecond regime, with pulsed electrons.

  6. Design and implementation of an optimal laser pulse front tilting scheme for ultrafast electron diffraction in reflection geometry with high temporal resolution

    PubMed Central

    Pennacchio, Francesco; Vanacore, Giovanni M.; Mancini, Giulia F.; Oppermann, Malte; Jayaraman, Rajeswari; Musumeci, Pietro; Baum, Peter; Carbone, Fabrizio

    2017-01-01

    Ultrafast electron diffraction is a powerful technique to investigate out-of-equilibrium atomic dynamics in solids with high temporal resolution. When diffraction is performed in reflection geometry, the main limitation is the mismatch in group velocity between the overlapping pump light and the electron probe pulses, which affects the overall temporal resolution of the experiment. A solution already available in the literature involved pulse front tilt of the pump beam at the sample, providing a sub-picosecond time resolution. However, in the reported optical scheme, the tilted pulse is characterized by a temporal chirp of about 1 ps at 1 mm away from the centre of the beam, which limits the investigation of surface dynamics in large crystals. In this paper, we propose an optimal tilting scheme designed for a radio-frequency-compressed ultrafast electron diffraction setup working in reflection geometry with 30 keV electron pulses containing up to 105 electrons/pulse. To characterize our scheme, we performed optical cross-correlation measurements, obtaining an average temporal width of the tilted pulse lower than 250 fs. The calibration of the electron-laser temporal overlap was obtained by monitoring the spatial profile of the electron beam when interacting with the plasma optically induced at the apex of a copper needle (plasma lensing effect). Finally, we report the first time-resolved results obtained on graphite, where the electron-phonon coupling dynamics is observed, showing an overall temporal resolution in the sub-500 fs regime. The successful implementation of this configuration opens the way to directly probe structural dynamics of low-dimensional systems in the sub-picosecond regime, with pulsed electrons. PMID:28713841

  7. Super-resolution nanofabrication with metal-ion doped hybrid material through an optical dual-beam approach

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cao, Yaoyu; Li, Xiangping; Gu, Min, E-mail: mgu@swin.edu.au

    2014-12-29

    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 formore » realizing functional nanostructures.« less

  8. Fundamental limits to superresolution fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Small, Alex

    2013-02-01

    Superresolution fluorescence microscopy techniques such as PALM, STORM, STED, and Structured Illumination Microscopy (SIM) enable imaging of live cells at nanometer resolution. The common theme in all of these techniques is that the diffraction limit is circumvented by controlling the states of fluorescent molecules. Although the samples are labeled very densely (i.e. with spacing much smaller than the Airy distance), not all of the molecules are emitting at the same time. Consequently, one does not encounter overlapping blurs. In the deterministic techniques (STED, SIM) the achievable resolution scales as the wavelength of light divided by the square root of the intensity of a beam used to control the fluorescent state. In the stochastic techniques (PALM, STORM), the achievable resolution scales as the wavelength of light divided by the square root of the number of photons collected. Although these limits arise from very different mechanisms (parabolic beam profiles for STED and SIM, statistics for PALM and STORM), in all cases the resolution scales inversely with the square root of a measure of the number of photons used in the experiment. We have developed a proof that this relationship between resolution and photon count is universal to techniques that control the states of fluorophores using classical light. Our proof encompasses linear and nonlinear optics, as well as computational post-processing techniques for extracting information beyond the diffraction limit. If there are techniques that can achieve a more efficient relationship between resolution and photon count, those techniques will require light exhibiting non-classical correlations.

  9. Biological imaging by soft X-ray diffraction microscopy

    NASA Astrophysics Data System (ADS)

    Shapiro, David

    We have developed a microscope for soft x-ray diffraction imaging of dry or frozen hydrated biological specimens. This lensless imaging system does not suffer from the resolution or specimen thickness limitations that other short wavelength microscopes experience. The microscope, currently situated at beamline 9.0.1 of the Advanced Light Source, can collect diffraction data to 12 nm resolution with 750 eV photons and 17 nm resolution with 520 eV photons. The specimen can be rotated with a precision goniometer through an angle of 160 degrees allowing for the collection of nearly complete three-dimensional diffraction data. The microscope is fully computer controlled through a graphical user interface and a scripting language automates the collection of both two-dimensional and three-dimensional data. Diffraction data from a freeze-dried dwarf yeast cell, Saccharomyces cerevisiae carrying the CLN3-1 mutation, was collected to 12 run resolution from 8 specimen orientations spanning a total rotation of 8 degrees. The diffraction data was phased using the difference map algorithm and the reconstructions provide real space images of the cell to 30 nm resolution from each of the orientations. The agreement of the different reconstructions provides confidence in the recovered, and previously unknown, structure and indicates the three dimensionality of the cell. This work represents the first imaging of the natural complex refractive contrast from a whole unstained cell by the diffraction microscopy method and has achieved a resolution superior to lens based x-ray tomographic reconstructions of similar specimens. Studies of the effects of exposure to large radiation doses were also carried out. It was determined that the freeze-dried cell suffers from an initial collapse, which is followed by a uniform, but slow, shrinkage. This structural damage to the cell is not accompanied by a diminished ability to see small features in the specimen. Preliminary measurements on frozen-hydrated yeast indicate that the frozen specimens do not exhibit these changes even with doses as high as 5 x 109 Gray.

  10. Accessing High Spatial Resolution in Astronomy Using Interference Methods

    ERIC Educational Resources Information Center

    Carbonel, Cyril; Grasset, Sébastien; Maysonnave, Jean

    2018-01-01

    In astronomy, methods such as direct imaging or interferometry-based techniques (Michelson stellar interferometry for example) are used for observations. A particular advantage of interferometry is that it permits greater spatial resolution compared to direct imaging with a single telescope, which is limited by diffraction owing to the aperture of…

  11. Coherent imaging at the diffraction limit

    PubMed Central

    Thibault, Pierre; Guizar-Sicairos, Manuel; Menzel, Andreas

    2014-01-01

    X-ray ptychography, a scanning coherent diffractive imaging technique, holds promise for imaging with dose-limited resolution and sensitivity. If the foreseen increase of coherent flux by orders of magnitude can be matched by additional technological and analytical advances, ptychography may approach imaging speeds familiar from full-field methods while retaining its inherently quantitative nature and metrological versatility. Beyond promises of high throughput, spectroscopic applications in three dimensions become feasible, as do measurements of sample dynamics through time-resolved imaging or careful characterization of decoherence effects. PMID:25177990

  12. Coherent imaging at the diffraction limit.

    PubMed

    Thibault, Pierre; Guizar-Sicairos, Manuel; Menzel, Andreas

    2014-09-01

    X-ray ptychography, a scanning coherent diffractive imaging technique, holds promise for imaging with dose-limited resolution and sensitivity. If the foreseen increase of coherent flux by orders of magnitude can be matched by additional technological and analytical advances, ptychography may approach imaging speeds familiar from full-field methods while retaining its inherently quantitative nature and metrological versatility. Beyond promises of high throughput, spectroscopic applications in three dimensions become feasible, as do measurements of sample dynamics through time-resolved imaging or careful characterization of decoherence effects.

  13. Indexing amyloid peptide diffraction from serial femtosecond crystallography: new algorithms for sparse patterns

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Brewster, Aaron S.; Sawaya, Michael R.; University of California, Los Angeles, CA 90095-1570

    2015-02-01

    Special methods are required to interpret sparse diffraction patterns collected from peptide crystals at X-ray free-electron lasers. Bragg spots can be indexed from composite-image powder rings, with crystal orientations then deduced from a very limited number of spot positions. Still diffraction patterns from peptide nanocrystals with small unit cells are challenging to index using conventional methods owing to the limited number of spots and the lack of crystal orientation information for individual images. New indexing algorithms have been developed as part of the Computational Crystallography Toolbox (cctbx) to overcome these challenges. Accurate unit-cell information derived from an aggregate data setmore » from thousands of diffraction patterns can be used to determine a crystal orientation matrix for individual images with as few as five reflections. These algorithms are potentially applicable not only to amyloid peptides but also to any set of diffraction patterns with sparse properties, such as low-resolution virus structures or high-throughput screening of still images captured by raster-scanning at synchrotron sources. As a proof of concept for this technique, successful integration of X-ray free-electron laser (XFEL) data to 2.5 Å resolution for the amyloid segment GNNQQNY from the Sup35 yeast prion is presented.« less

  14. Efficient photonic reformatting of celestial light for diffraction-limited spectroscopy

    NASA Astrophysics Data System (ADS)

    MacLachlan, D. G.; Harris, R. J.; Gris-Sánchez, I.; Morris, T. J.; Choudhury, D.; Gendron, E.; Basden, A. G.; Spaleniak, I.; Arriola, A.; Birks, T. A.; Allington-Smith, J. R.; Thomson, R. R.

    2017-02-01

    The spectral resolution of a dispersive astronomical spectrograph is limited by the trade-off between throughput and the width of the entrance slit. Photonic guided wave transitions have been proposed as a route to bypass this trade-off, by enabling the efficient reformatting of incoherent seeing-limited light collected by the telescope into a linear array of single modes: a pseudo-slit which is highly multimode in one axis but diffraction-limited in the dispersion axis of the spectrograph. It is anticipated that the size of a single-object spectrograph fed with light in this manner would be essentially independent of the telescope aperture size. A further anticipated benefit is that such spectrographs would be free of `modal noise', a phenomenon that occurs in high-resolution multimode fibre-fed spectrographs due to the coherent nature of the telescope point spread function (PSF). We seek to address these aspects by integrating a multicore fibre photonic lantern with an ultrafast laser inscribed three-dimensional waveguide interconnect to spatially reformat the modes within the PSF into a diffraction-limited pseudo-slit. Using the CANARY adaptive optics (AO) demonstrator on the William Herschel Telescope, and 1530 ± 80 nm stellar light, the device exhibits a transmission of 47-53 per cent depending upon the mode of AO correction applied. We also show the advantage of using AO to couple light into such a device by sampling only the core of the CANARY PSF. This result underscores the possibility that a fully optimized guided-wave device can be used with AO to provide efficient spectroscopy at high spectral resolution.

  15. Theoretical modeling and evaluation of the axial resolution of the adaptive optics scanning laser ophthalmoscope.

    PubMed

    Venkateswaran, Krishnakumar; Roorda, Austin; Romero-Borja, Fernando

    2004-01-01

    We present axial resolution calculated using a mathematical model of the adaptive optics scanning laser ophthalmoscope (AOSLO). The peak intensity and the width of the axial intensity response are computed with the residual Zernike coefficients after the aberrations are corrected using adaptive optics for eight subjects and compared with the axial resolution of a diffraction-limited eye. The AOSLO currently uses a confocal pinhole that is 80 microm, or 3.48 times the width of the Airy disk radius of the collection optics, and projects to 7.41 microm on the retina. For this pinhole, the axial resolution of a diffraction-limited system is 114 microm and the computed axial resolution varies between 120 and 146 microm for the human subjects included in this study. The results of this analysis indicate that to improve axial resolution, it is best to reduce the pinhole size. The resulting reduction in detected light may demand, however, a more sophisticated adaptive optics system. The study also shows that imaging systems with large pinholes are relatively insensitive to misalignment in the lateral positioning of the confocal pinhole. However, when small pinholes are used to maximize resolution, alignment becomes critical. ( c) 2004 Society of Photo-Optical Instrumentation Engineers.

  16. The Adaptive Optics Lucky Imager: Diffraction limited imaging at visible wavelengths with large ground-based telescopes

    NASA Astrophysics Data System (ADS)

    Crass, Jonathan; Mackay, Craig; King, David; Rebolo-López, Rafael; Labadie, Lucas; Puga, Marta; Oscoz, Alejandro; González Escalera, Victor; Pérez Garrido, Antonio; López, Roberto; Pérez-Prieto, Jorge; Rodríguez-Ramos, Luis; Velasco, Sergio; Villó, Isidro

    2015-01-01

    One of the continuing challenges facing astronomers today is the need to obtain ever higher resolution images of the sky. Whether studying nearby crowded fields or distant objects, with increased resolution comes the ability to probe systems in more detail and advance our understanding of the Universe. Obtaining these high-resolution images at visible wavelengths however has previously been limited to the Hubble Space Telescope (HST) due to atmospheric effects limiting the spatial resolution of ground-based telescopes to a fraction of their potential. With HST now having a finite lifespan, it is prudent to investigate other techniques capable of providing these kind of observations from the ground. Maintaining this capability is one of the goals of the Adaptive Optics Lucky Imager (AOLI).Achieving the highest resolutions requires the largest telescope apertures, however, this comes at the cost of increased atmospheric distortion. To overcome these atmospheric effects, there are two main techniques employed today: adaptive optics (AO) and lucky imaging. These techniques individually are unable to provide diffraction limited imaging in the visible on large ground-based telescopes; AO currently only works at infrared wavelengths while lucky imaging reduces in effectiveness on telescopes greater than 2.5 metres in diameter. The limitations of both techniques can be overcome by combing them together to provide diffraction limited imaging at visible wavelengths on the ground.The Adaptive Optics Lucky Imager is being developed as a European collaboration and combines AO and lucky imaging in a dedicated instrument for the first time. Initially for use on the 4.2 metre William Herschel Telescope, AOLI uses a low-order adaptive optics system to reduce the effects of atmospheric turbulence before imaging with a lucky imaging based science detector. The AO system employs a novel type of wavefront sensor, the non-linear Curvature Wavefront Sensor (nlCWFS) which provides significant sky-coverage using natural guide-stars alone.Here we present an overview of the instrument design, results from the first on-sky and laboratory testing and on-going development work of the instrument and its adaptive optics system.

  17. Evanescent Properties of Optical Diffraction from 2-Dimensional Hexagonal Photonic Crystals and Their Sensor Applications.

    PubMed

    Liao, Yu-Yang; Chen, Yung-Tsan; Chen, Chien-Chun; Huang, Jian-Jang

    2018-04-03

    The sensitivity of traditional diffraction grating sensors is limited by the spatial resolution of the measurement setup. Thus, a large space is required to improve sensor performance. Here, we demonstrate a compact hexagonal photonic crystal (PhC) optical sensor with high sensitivity. PhCs are able to diffract optical beams to various angles in azimuthal space. The critical wavelength that satisfies the phase matching or becomes evanescent was used to benchmark the refractive index of a target analyte applied on a PhC sensor. Using a glucose solution as an example, our sensor demonstrated very high sensitivity and a low limit of detection. This shows that the diffraction mechanism of hexagonal photonic crystals can be used for sensors when compact size is a concern.

  18. Far Infrared Imaging Spectrometer for Large Aperture Infrared Telescope System

    DTIC Science & Technology

    1985-12-01

    resolution Fabry - Perot spectrometer (103 < Resolution < 104) for wavelengths from about 50 to 200 micrometer, employing extended field diffraction limited...photo- metry. The Naval Research Laboratory will provide a high resolution Far Infrared Imaging Spectrometer (FIRIS) using Fabry - Perot techniques in...detectors to provide spatial information. The Fabry - Perot uses electromagnetic coil displacement drivers with a lead screw drive to obtain parallel

  19. Translation position determination in ptychographic coherent diffraction imaging.

    PubMed

    Zhang, Fucai; Peterson, Isaac; Vila-Comamala, Joan; Diaz, Ana; Berenguer, Felisa; Bean, Richard; Chen, Bo; Menzel, Andreas; Robinson, Ian K; Rodenburg, John M

    2013-06-03

    Accurate knowledge of translation positions is essential in ptychography to achieve a good image quality and the diffraction limited resolution. We propose a method to retrieve and correct position errors during the image reconstruction iterations. Sub-pixel position accuracy after refinement is shown to be achievable within several tens of iterations. Simulation and experimental results for both optical and X-ray wavelengths are given. The method improves both the quality of the retrieved object image and relaxes the position accuracy requirement while acquiring the diffraction patterns.

  20. High-resolution structure of viruses from random diffraction snapshots

    PubMed Central

    Hosseinizadeh, A.; Schwander, P.; Dashti, A.; Fung, R.; D'Souza, R. M.; Ourmazd, A.

    2014-01-01

    The advent of the X-ray free-electron laser (XFEL) has made it possible to record diffraction snapshots of biological entities injected into the X-ray beam before the onset of radiation damage. Algorithmic means must then be used to determine the snapshot orientations and thence the three-dimensional structure of the object. Existing Bayesian approaches are limited in reconstruction resolution typically to 1/10 of the object diameter, with the computational expense increasing as the eighth power of the ratio of diameter to resolution. We present an approach capable of exploiting object symmetries to recover three-dimensional structure to high resolution, and thus reconstruct the structure of the satellite tobacco necrosis virus to atomic level. Our approach offers the highest reconstruction resolution for XFEL snapshots to date and provides a potentially powerful alternative route for analysis of data from crystalline and nano-crystalline objects. PMID:24914154

  1. High-resolution structure of viruses from random diffraction snapshots.

    PubMed

    Hosseinizadeh, A; Schwander, P; Dashti, A; Fung, R; D'Souza, R M; Ourmazd, A

    2014-07-17

    The advent of the X-ray free-electron laser (XFEL) has made it possible to record diffraction snapshots of biological entities injected into the X-ray beam before the onset of radiation damage. Algorithmic means must then be used to determine the snapshot orientations and thence the three-dimensional structure of the object. Existing Bayesian approaches are limited in reconstruction resolution typically to 1/10 of the object diameter, with the computational expense increasing as the eighth power of the ratio of diameter to resolution. We present an approach capable of exploiting object symmetries to recover three-dimensional structure to high resolution, and thus reconstruct the structure of the satellite tobacco necrosis virus to atomic level. Our approach offers the highest reconstruction resolution for XFEL snapshots to date and provides a potentially powerful alternative route for analysis of data from crystalline and nano-crystalline objects.

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

    NASA Astrophysics Data System (ADS)

    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.

  3. SIL-STED microscopy technique enhancing super-resolution of fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Park, No-Cheol; Lim, Geon; Lee, Won-sup; Moon, Hyungbae; Choi, Guk-Jong; Park, Young-Pil

    2017-08-01

    We have characterized a new type STED microscope which combines a high numerical aperture (NA) optical head with a solid immersion lens (SIL), and we call it as SIL-STED microscope. The advantage of a SIL-STED microscope is that its high NA of the SIL makes it superior to a general STED microscope in lateral resolution, thus overcoming the optical diffraction limit at the macromolecular level and enabling advanced super-resolution imaging of cell surface or cell membrane structure and function Do. This study presents the first implementation of higher NA illumination in a STED microscope limiting the fluorescence lateral resolution to about 40 nm. The refractive index of the SIL which is made of material KTaO3 is about 2.23 and 2.20 at a wavelength of 633 nm and 780 nm which are used for excitation and depletion in STED imaging, respectively. Based on the vector diffraction theory, the electric field focused by the SILSTED microscope is numerically calculated so that the numerical results of the point dispersion function of the microscope and the expected resolution could be analyzed. For further investigation, fluorescence imaging of nano size fluorescent beads is fulfilled to show improved performance of the technique.

  4. Evanescent-Wave Filtering in Images Using Remote Terahertz Structured Illumination

    NASA Astrophysics Data System (ADS)

    Flammini, M.; Pontecorvo, E.; Giliberti, V.; Rizza, C.; Ciattoni, A.; Ortolani, M.; DelRe, E.

    2017-11-01

    Imaging with structured illumination allows for the retrieval of subwavelength features of an object by conversion of evanescent waves into propagating waves. In conditions in which the object plane and the structured-illumination plane do not coincide, this conversion process is subject to progressive filtering of the components with high spatial frequency when the distance between the two planes increases, until the diffraction-limited lateral resolution is restored when the distance exceeds the extension of evanescent waves. We study the progressive filtering of evanescent waves by developing a remote super-resolution terahertz imaging system operating at a wavelength λ =1.00 mm , based on a freestanding knife edge and a reflective confocal terahertz microscope. In the images recorded with increasing knife-edge-to-object-plane distance, we observe the transition from a super-resolution of λ /17 ≃60 μ m to the diffraction-limited lateral resolution of Δ x ≃λ expected for our confocal microscope. The extreme nonparaxial conditions are analyzed in detail, exploiting the fact that, in the terahertz frequency range, the knife edge can be positioned at a variable subwavelength distance from the object plane. Electromagnetic simulations of radiation scattering by the knife edge reproduce the experimental super-resolution achieved.

  5. Absolute angular encoder based on optical diffraction

    NASA Astrophysics Data System (ADS)

    Wu, Jian; Zhou, Tingting; Yuan, Bo; Wang, Liqiang

    2015-08-01

    A new encoding method for absolute angular encoder based on optical diffraction was proposed in the present study. In this method, an encoder disc is specially designed that a series of elements are uniformly spaced in one circle and each element is consisted of four diffraction gratings, which are tilted in the directions of 30°, 60°, -60° and -30°, respectively. The disc is illuminated by a coherent light and the diffractive signals are received. The positions of diffractive spots are used for absolute encoding and their intensities are for subdivision, which is different from the traditional optical encoder based on transparent/opaque binary principle. Since the track's width in the disc is not limited in the diffraction pattern, it provides a new way to solve the contradiction between the size and resolution, which is good for minimization of encoder. According to the proposed principle, the diffraction pattern disc with a diameter of 40 mm was made by lithography in the glass substrate. A prototype of absolute angular encoder with a resolution of 20" was built up. Its maximum error was tested as 78" by comparing with a small angle measuring system based on laser beam deflection.

  6. A large area high resolution imaging detector for fast atom diffraction

    NASA Astrophysics Data System (ADS)

    Lupone, Sylvain; Soulisse, Pierre; Roncin, Philippe

    2018-07-01

    We describe a high resolution imaging detector based on a single 80 mm micro-channel-plate (MCP) and a phosphor screen mounted on a UHV flange of only 100 mm inner diameter. It relies on standard components and we describe its performance with one or two MCPs. A resolution of 80 μm rms is observed on the beam profile. At low count rate, individual impact can be pinpointed with few μm accuracy but the resolution is probably limited by the MCP channel diameter. The detector has been used to record the diffraction of fast atoms at grazing incidence on crystal surfaces (GIFAD), a technique probing the electronic density of the topmost layer only. The detector was also used to record the scattering profile during azimuthal scan of the crystal to produce triangulation curves revealing the surface crystallographic directions of molecular layers. It should also be compatible with reflection high energy electron (RHEED) experiment when fragile surfaces require a low exposure to the electron beam. The discussions on the mode of operation specific to diffraction experiments apply also to commercial detectors.

  7. Compact high-resolution spectrographs for large and extremely large telescopes: using the diffraction limit

    NASA Astrophysics Data System (ADS)

    Robertson, J. Gordon; Bland-Hawthorn, Joss

    2012-09-01

    As telescopes get larger, the size of a seeing-limited spectrograph for a given resolving power becomes larger also, and for ELTs the size will be so great that high resolution instruments of simple design will be infeasible. Solutions include adaptive optics (but not providing full correction for short wavelengths) or image slicers (which give feasible but still large instruments). Here we develop the solution proposed by Bland-Hawthorn and Horton: the use of diffraction-limited spectrographs which are compact even for high resolving power. Their use is made possible by the photonic lantern, which splits a multi-mode optical fiber into a number of single-mode fibers. We describe preliminary designs for such spectrographs, at a resolving power of R ~ 50,000. While they are small and use relatively simple optics, the challenges are to accommodate the longest possible fiber slit (hence maximum number of single-mode fibers in one spectrograph) and to accept the beam from each fiber at a focal ratio considerably faster than for most spectrograph collimators, while maintaining diffraction-limited imaging quality. It is possible to obtain excellent performance despite these challenges. We also briefly consider the number of such spectrographs required, which can be reduced by full or partial adaptive optics correction, and/or moving towards longer wavelengths.

  8. Transverse Coherence Limited Coherent Diffraction Imaging using a Molybdenum Soft X-ray Laser Pumped at Moderate Pump Energies.

    PubMed

    Zürch, M; Jung, R; Späth, C; Tümmler, J; Guggenmos, A; Attwood, D; Kleineberg, U; Stiel, H; Spielmann, C

    2017-07-13

    Coherent diffraction imaging (CDI) in the extreme ultraviolet has become an important tool for nanoscale investigations. Laser-driven high harmonic generation (HHG) sources allow for lab scale applications such as cancer cell classification and phase-resolved surface studies. HHG sources exhibit excellent coherence but limited photon flux due poor conversion efficiency. In contrast, table-top soft X-ray lasers (SXRL) feature excellent temporal coherence and extraordinary high flux at limited transverse coherence. Here, the performance of a SXRL pumped at moderate pump energies is evaluated for CDI and compared to a HHG source. For CDI, a lower bound for the required mutual coherence factor of |μ 12 | ≥ 0.75 is found by comparing a reconstruction with fixed support to a conventional characterization using double slits. A comparison of the captured diffraction signals suggests that SXRLs have the potential for imaging micron scale objects with sub-20 nm resolution in orders of magnitude shorter integration time compared to a conventional HHG source. Here, the low transverse coherence diameter limits the resolution to approximately 180 nm. The extraordinary high photon flux per laser shot, scalability towards higher repetition rate and capability of seeding with a high harmonic source opens a route for higher performance nanoscale imaging systems based on SXRLs.

  9. Super-resolution optical telescopes with local light diffraction shrinkage

    PubMed Central

    Wang, Changtao; Tang, Dongliang; Wang, Yanqin; Zhao, Zeyu; Wang, Jiong; Pu, Mingbo; Zhang, Yudong; Yan, Wei; Gao, Ping; Luo, Xiangang

    2015-01-01

    Suffering from giant size of objective lenses and infeasible manipulations of distant targets, telescopes could not seek helps from present super-resolution imaging, such as scanning near-field optical microscopy, perfect lens and stimulated emission depletion microscopy. In this paper, local light diffraction shrinkage associated with optical super-oscillatory phenomenon is proposed for real-time and optically restoring super-resolution imaging information in a telescope system. It is found that fine target features concealed in diffraction-limited optical images of a telescope could be observed in a small local field of view, benefiting from a relayed metasurface-based super-oscillatory imaging optics in which some local Fourier components beyond the cut-off frequency of telescope could be restored. As experimental examples, a minimal resolution to 0.55 of Rayleigh criterion is obtained, and imaging complex targets and large targets by superimposing multiple local fields of views are demonstrated as well. This investigation provides an access for real-time, incoherent and super-resolution telescopes without the manipulation of distant targets. More importantly, it gives counterintuitive evidence to the common knowledge that relayed optics could not deliver more imaging details than objective systems. PMID:26677820

  10. High spatial resolution with zoomable saw-tooth refractive lenses?

    NASA Astrophysics Data System (ADS)

    Jark, Werner

    2011-09-01

    Refractive x-ray lenses can be assembled from two opposing saw-tooth structures, when they are inclined with respect to each other and almost touch at one end. An incident plane wave will then traverse a varying number of triangular prisms, which direct the beam towards the optical axis and focus it. Optically speaking the plane wave traverses a parabolic lens profile, which is approximated by trapezoidal segments. The parabolic profile will focus ideally, when a lens can be discussed in the "thin lens" approximation. Now the saw-tooth refractive lens is found to be too "thick". The residual aberrations limit the focusing capability to just submicrometer focusing, significantly above the limit in diffraction limited focusing. It is shown that the aberrations can be removed by introducing a variation into the originally constant saw-tooth angle. After this modification the lens can be operated in the diffraction limited regime. Spot sizes even below 0.1 micrometer are then feasible. This performance in terms of spatial resolution is found to be limited to focusing to microspots and is not available, when the saw-tooth refractive lens is used in an imaging setup. In this case the spatial resolution deteriorates rapidly with increasing off axis distance of the object to be imaged.

  11. A versatile diffractive maskless lithography for single-shot and serial microfabrication.

    PubMed

    Jenness, Nathan J; Hill, Ryan T; Hucknall, Angus; Chilkoti, Ashutosh; Clark, Robert L

    2010-05-24

    We demonstrate a diffractive maskless lithographic system that is capable of rapidly performing both serial and single-shot micropatterning. Utilizing the diffractive properties of phase holograms displayed on a spatial light modulator, arbitrary intensity distributions were produced to form two and three dimensional micropatterns/structures in a variety of substrates. A straightforward graphical user interface was implemented to allow users to load templates and change patterning modes within the span of a few minutes. A minimum resolution of approximately 700 nm is demonstrated for both patterning modes, which compares favorably to the 232 nm resolution limit predicted by the Rayleigh criterion. The presented method is rapid and adaptable, allowing for the parallel fabrication of microstructures in photoresist as well as the fabrication of protein microstructures that retain functional activity.

  12. Near-field microscopy with a microfabricated solid immersion lens

    NASA Astrophysics Data System (ADS)

    Fletcher, Daniel Alden

    2001-07-01

    Diffraction of focused light prevents optical microscopes from resolving features in air smaller than half the wavelength, λ Spatial resolution can be improved by passing light through a sub-wavelength metal aperture scanned close to a sample, but aperture-based probes suffer from low optical throughput, typically below 10-4. An alternate and more efficient technique is solid immersion microscopy in which light is focused through a high refractive index Solid Immersion Lens (SIL). This work describes the fabrication, modeling, and use of a microfabricated SIL to obtain spatial resolution better than the diffraction limit in air with high optical throughput for infrared applications. SILs on the order of 10 μm in diameter are fabricated from single-crystal silicon and integrated onto silicon cantilevers with tips for scanning. We measure a focused spot size of λ/5 with optical throughput better than 10-1 at a wavelength of λ = 9.3 μm. Spatial resolution is improved to λ/10 with metal apertures fabricated directly on the tip of the silicon SIL. Microlenses have reduced spherical aberration and better transparency than large lenses but cannot be made arbitrarily small and still focus. We model the advantages and limitations of focusing in lenses close to the wavelength in diameter using an extension of Mie theory. We also investigate a new contrast mechanism unique to microlenses resulting from the decrease in field-of-view with lens diameter. This technique is shown to achieve λ/4 spatial resolution. We explore applications of the microfabricated silicon SIL for high spatial resolution thermal microscopy and biological spectroscopy. Thermal radiation is collected through the SIL from a heated surface with spatial resolution four times better than that of a diffraction- limited infrared microscope. Using a Fourier-transform infrared spectrometer, we observe absorption peaks in bacteria cells positioned at the focus of the silicon SIL.

  13. Resolution factors in edgeline holography.

    PubMed

    Trolinger, J D; Gee, T H

    1971-06-01

    When an in-line Fresnel hologram of an object such as a projectile in flight is made, the reconstruction comprises an image of the outside edge of the object superimposed upon a Fresnel diffraction pattern of the edge and an unmodulated portion of the reconstruction beam. When the reconstructed image is bandpass filtered, the only remaining significant contribution is that of a diffraction pattern which is symmetrical about an edgeline gaussian image of the object. The present paper discusses the application of this type of holography in accurately locating the edge of a large dynamic object, the position of which is not accurately known in any dimension. A theoretical and experimental analysis was performed to study the effects of motion, hologram size, film type, and practical limitations upon the attainable resolution in the reconstructed image. The bandlimiting effect of motion is used to relate the motion effected resolution limit of holography to that of photography. The study shows that an edgeline can be accurately located even at high velocity normal to the edge.

  14. Experimental assessment and analysis of super-resolution in fluorescence microscopy based on multiple-point spread function fitting of spectrally demultiplexed images

    NASA Astrophysics Data System (ADS)

    Nishimura, Takahiro; Kimura, Hitoshi; Ogura, Yusuke; Tanida, Jun

    2018-06-01

    This paper presents an experimental assessment and analysis of super-resolution microscopy based on multiple-point spread function fitting of spectrally demultiplexed images using a designed DNA structure as a test target. For the purpose, a DNA structure was designed to have binding sites at a certain interval that is smaller than the diffraction limit. The structure was labeled with several types of quantum dots (QDs) to acquire their spatial information as spectrally encoded images. The obtained images are analyzed with a point spread function multifitting algorithm to determine the QD locations that indicate the binding site positions. The experimental results show that the labeled locations can be observed beyond the diffraction-limited resolution using three-colored fluorescence images that were obtained with a confocal fluorescence microscope. Numerical simulations show that labeling with eight types of QDs enables the positions aligned at 27.2-nm pitches on the DNA structure to be resolved with high accuracy.

  15. Facing the phase problem in Coherent Diffractive Imaging via Memetic Algorithms.

    PubMed

    Colombo, Alessandro; Galli, Davide Emilio; De Caro, Liberato; Scattarella, Francesco; Carlino, Elvio

    2017-02-09

    Coherent Diffractive Imaging is a lensless technique that allows imaging of matter at a spatial resolution not limited by lens aberrations. This technique exploits the measured diffraction pattern of a coherent beam scattered by periodic and non-periodic objects to retrieve spatial information. The diffracted intensity, for weak-scattering objects, is proportional to the modulus of the Fourier Transform of the object scattering function. Any phase information, needed to retrieve its scattering function, has to be retrieved by means of suitable algorithms. Here we present a new approach, based on a memetic algorithm, i.e. a hybrid genetic algorithm, to face the phase problem, which exploits the synergy of deterministic and stochastic optimization methods. The new approach has been tested on simulated data and applied to the phasing of transmission electron microscopy coherent electron diffraction data of a SrTiO 3 sample. We have been able to quantitatively retrieve the projected atomic potential, and also image the oxygen columns, which are not directly visible in the relevant high-resolution transmission electron microscopy images. Our approach proves to be a new powerful tool for the study of matter at atomic resolution and opens new perspectives in those applications in which effective phase retrieval is necessary.

  16. Limit characteristics of digital optoelectronic processor

    NASA Astrophysics Data System (ADS)

    Kolobrodov, V. G.; Tymchik, G. S.; Kolobrodov, M. S.

    2018-01-01

    In this article, the limiting characteristics of a digital optoelectronic processor are explored. The limits are defined by diffraction effects and a matrix structure of the devices for input and output of optical signals. The purpose of a present research is to optimize the parameters of the processor's components. The developed physical and mathematical model of DOEP allowed to establish the limit characteristics of the processor, restricted by diffraction effects and an array structure of the equipment for input and output of optical signals, as well as to optimize the parameters of the processor's components. The diameter of the entrance pupil of the Fourier lens is determined by the size of SLM and the pixel size of the modulator. To determine the spectral resolution, it is offered to use a concept of an optimum phase when the resolved diffraction maxima coincide with the pixel centers of the radiation detector.

  17. Subwavelength resolution from multilayered structure (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Cheng, Bo Han; Jen, Yi-Jun; Liu, Wei-Chih; Lin, Shan-wen; Lan, Yung-Chiang; Tsai, Din Ping

    2016-10-01

    Breaking optical diffraction limit is one of the most important issues needed to be overcome for the demand of high-density optoelectronic components. Here, a multilayered structure which consists of alternating semiconductor and dielectric layers for breaking optical diffraction limitation at THz frequency region are proposed and analyzed. We numerically demonstrate that such multilayered structure not only can act as a hyperbolic metamaterial but also a birefringence material via the control of the external temperature (or magnetic field). A practical approach is provided to control all the diffraction signals toward a specific direction by using transfer matrix method and effective medium theory. Numerical calculations and computer simulation (based on finite element method, FEM) are carried out, which agree well with each other. The temperature (or magnetic field) parameter can be tuned to create an effective material with nearly flat isofrequency feature to transfer (project) all the k-space signals excited from the object to be resolved to the image plane. Furthermore, this multilayered structure can resolve subwavelength structures at various incident THz light sources simultaneously. In addition, the resolution power for a fixed operating frequency also can be tuned by only changing the magnitude of external magnetic field. Such a device provides a practical route for multi-functional material, photolithography and real-time super-resolution image.

  18. Relativistic electron diffraction at the UCLA Pegasus photoinjector laboratory.

    PubMed

    Musumeci, P; Moody, J T; Scoby, C M

    2008-10-01

    Electron diffraction holds the promise to yield real-time resolution of atomic motion in an easily accessible environment like a university laboratory at a fraction of the cost of fourth-generation X-ray sources. Currently the limit in time-resolution for conventional electron diffraction is set by how short an electron pulse can be made. A very promising solution to maintain the highest possible beam intensity without excessive pulse broadening from space charge effects is to increase the electron energy to the MeV level where relativistic effects significantly reduce the space charge forces. Rf photoinjectors can in principle deliver up to 10(7)-10(8) electrons packed in bunches of approximately 100-fs length, allowing an unprecedented time resolution and enabling the study of irreversible phenomena by single-shot diffraction patterns. The use of rf photoinjectors as sources for ultrafast electron diffraction has been recently at the center of various theoretical and experimental studies. The UCLA Pegasus laboratory, commissioned in early 2007 as an advanced photoinjector facility, is the only operating system in the country, which has recently demonstrated electron diffraction using a relativistic beam from an rf photoinjector. Due to the use of a state-of-the-art ultrashort photoinjector driver laser system, the beam has been measured to be sub-100-fs long, at least a factor of 5 better than what measured in previous relativistic electron diffraction setups. Moreover, diffraction patterns from various metal targets (titanium and aluminum) have been obtained using the Pegasus beam. One of the main laboratory goals in the near future is to fully develop the rf photoinjector-based ultrafast electron diffraction technique with particular attention to the optimization of the working point of the photoinjector in a low-charge ultrashort pulse regime, and to the development of suitable beam diagnostics.

  19. Ptychographic overlap constraint errors and the limits of their numerical recovery using conjugate gradient descent methods.

    PubMed

    Tripathi, Ashish; McNulty, Ian; Shpyrko, Oleg G

    2014-01-27

    Ptychographic coherent x-ray diffractive imaging is a form of scanning microscopy that does not require optics to image a sample. A series of scanned coherent diffraction patterns recorded from multiple overlapping illuminated regions on the sample are inverted numerically to retrieve its image. The technique recovers the phase lost by detecting the diffraction patterns by using experimentally known constraints, in this case the measured diffraction intensities and the assumed scan positions on the sample. The spatial resolution of the recovered image of the sample is limited by the angular extent over which the diffraction patterns are recorded and how well these constraints are known. Here, we explore how reconstruction quality degrades with uncertainties in the scan positions. We show experimentally that large errors in the assumed scan positions on the sample can be numerically determined and corrected using conjugate gradient descent methods. We also explore in simulations the limits, based on the signal to noise of the diffraction patterns and amount of overlap between adjacent scan positions, of just how large these errors can be and still be rendered tractable by this method.

  20. High Sensitivity, High Angular Resolution Far-infrared Photometry from the KAO

    NASA Technical Reports Server (NTRS)

    Lester, D.; Harvey, P. M.; Wilking, B. A.; Joy, M.

    1984-01-01

    Most of the luminosity of embedded sources is reemitted in the far-infrared continuum. Measurements in the far-infrared are essential to understand the energetics of the interstellar medium, and of star formation regions in particular. Measurements from the KAO, are made in diffraction limited beams that sample a spatial scale considerably smaller than that given by IRAS. The KAO instrument technology has matured to the point that the single scan limiting flux of IRAS at 100 micro can be reached in a diffraction limited beam in a single typical KAO observing leg. The far-infrared photometer system and selections of recent observations are presented.

  1. Pupil-segmentation-based adaptive optics for microscopy

    NASA Astrophysics Data System (ADS)

    Ji, Na; Milkie, Daniel E.; Betzig, Eric

    2011-03-01

    Inhomogeneous optical properties of biological samples make it difficult to obtain diffraction-limited resolution in depth. Correcting the sample-induced optical aberrations needs adaptive optics (AO). However, the direct wavefront-sensing approach commonly used in astronomy is not suitable for most biological samples due to their strong scattering of light. We developed an image-based AO approach that is insensitive to sample scattering. By comparing images of the sample taken with different segments of the pupil illuminated, local tilt in the wavefront is measured from image shift. The aberrated wavefront is then obtained either by measuring the local phase directly using interference or with phase reconstruction algorithms similar to those used in astronomical AO. We implemented this pupil-segmentation-based approach in a two-photon fluorescence microscope and demonstrated that diffraction-limited resolution can be recovered from nonbiological and biological samples.

  2. Dynamic full-field infrared imaging with multiple synchrotron beams

    PubMed Central

    Stavitski, Eli; Smith, Randy J.; Bourassa, Megan W.; Acerbo, Alvin S.; Carr, G. L.; Miller, Lisa M.

    2013-01-01

    Microspectroscopic imaging in the infrared (IR) spectral region allows for the examination of spatially resolved chemical composition on the microscale. More than a decade ago, it was demonstrated that diffraction limited spatial resolution can be achieved when an apertured, single pixel IR microscope is coupled to the high brightness of a synchrotron light source. Nowadays, many IR microscopes are equipped with multi-pixel Focal Plane Array (FPA) detectors, which dramatically improve data acquisition times for imaging large areas. Recently, progress been made toward efficiently coupling synchrotron IR beamlines to multi-pixel detectors, but they utilize expensive and highly customized optical schemes. Here we demonstrate the development and application of a simple optical configuration that can be implemented on most existing synchrotron IR beamlines in order to achieve full-field IR imaging with diffraction-limited spatial resolution. Specifically, the synchrotron radiation fan is extracted from the bending magnet and split into four beams that are combined on the sample, allowing it to fill a large section of the FPA. With this optical configuration, we are able to oversample an image by more than a factor of two, even at the shortest wavelengths, making image restoration through deconvolution algorithms possible. High chemical sensitivity, rapid acquisition times, and superior signal-to-noise characteristics of the instrument are demonstrated. The unique characteristics of this setup enabled the real time study of heterogeneous chemical dynamics with diffraction-limited spatial resolution for the first time. PMID:23458231

  3. Quantum enhanced superresolution microscopy (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Oron, Dan; Tenne, Ron; Israel, Yonatan; Silberberg, Yaron

    2017-02-01

    Far-field optical microscopy beyond the Abbe diffraction limit, making use of nonlinear excitation (e.g. STED), or temporal fluctuations in fluorescence (PALM, STORM, SOFI) is already a reality. In contrast, overcoming the diffraction limit using non-classical properties of light is very difficult to achieve due to the fragility of quantum states of light. Here, we experimentally demonstrate superresolution microscopy based on quantum properties of light naturally emitted by fluorophores used as markers in fluorescence microscopy. Our approach is based on photon antibunching, the tendency of fluorophores to emit photons one by one rather than in bursts. Although a distinctively quantum phenomenon, antibunching is readily observed in most common fluorophores even at room temperature. This nonclassical resource can be utilized directly to enhance the imaging resolution, since the non-classical far-field intensity correlations induced by antibunching carry high spatial frequency information on the spatial distribution of emitters. Detecting photon statistics simultaneously in the entire field of view, we were able to detect non-classical correlations of the second and third order, and reconstructed images with resolution significantly beyond the diffraction limit. Alternatively, we demonstrate the utilization of antibunching for augmenting the capabilities of localization-based superresolution imaging in the presence of multiple emitters, using a novel detector comprised of an array of single photon detectors connected to a densely packed fiber bundle. These features allow us to enhance the spatial and temporal resolution with which multiple emitters can be imaged compared with other techniques that rely on CCD cameras.

  4. Mission Concepts for High-Resolution Solar Imaging with a Photon Sieve

    NASA Astrophysics Data System (ADS)

    Rabin, Douglas M.; Davila, Joseph; Daw, Adrian N.; Denis, Kevin L.; Novo-Gradac, Anne-Marie; Shah, Neerav; Widmyer, Thomas R.

    2017-08-01

    The best EUV coronal imagers are unable to probe the expected energy dissipation scales of the solar corona (<100 km) because conventional optics cannot be figured to near diffraction-limited accuracy at these wavelengths. Davila (2011) has proposed that a photon sieve, a diffractive imaging element similar to a Fresnel zone plate, provides a technically feasible path to the required angular resolution. We have produced photon sieves as large as 80 mm clear aperture. We discuss laboratory measurements of these devices and the path to larger apertures. The focal length of a sieve with high EUV resolution is at least 10 m. Options for solar imaging with such a sieve include a sounding rocket, a single spacecraft with a deployed boom, and two spacecraft flying in precise formation.

  5. Indexing amyloid peptide diffraction from serial femtosecond crystallography: New algorithms for sparse patterns

    DOE PAGES

    Brewster, Aaron S.; Sawaya, Michael R.; Rodriguez, Jose; ...

    2015-01-23

    Still diffraction patterns from peptide nanocrystals with small unit cells are challenging to index using conventional methods owing to the limited number of spots and the lack of crystal orientation information for individual images. New indexing algorithms have been developed as part of the Computational Crystallography Toolbox( cctbx) to overcome these challenges. Accurate unit-cell information derived from an aggregate data set from thousands of diffraction patterns can be used to determine a crystal orientation matrix for individual images with as few as five reflections. These algorithms are potentially applicable not only to amyloid peptides but also to any set ofmore » diffraction patterns with sparse properties, such as low-resolution virus structures or high-throughput screening of still images captured by raster-scanning at synchrotron sources. As a proof of concept for this technique, successful integration of X-ray free-electron laser (XFEL) data to 2.5 Å resolution for the amyloid segment GNNQQNY from the Sup35 yeast prion is presented.« less

  6. Example-Based Super-Resolution Fluorescence Microscopy.

    PubMed

    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.

  7. Molecular Switch for Sub-Diffraction Laser Lithography by Photoenol Intermediate-State Cis-Trans Isomerization.

    PubMed

    Mueller, Patrick; Zieger, Markus M; Richter, Benjamin; Quick, Alexander S; Fischer, Joachim; Mueller, Jonathan B; Zhou, Lu; Nienhaus, Gerd Ulrich; Bastmeyer, Martin; Barner-Kowollik, Christopher; Wegener, Martin

    2017-06-27

    Recent developments in stimulated-emission depletion (STED) microscopy have led to a step change in the achievable resolution and allowed breaking the diffraction limit by large factors. The core principle is based on a reversible molecular switch, allowing for light-triggered activation and deactivation in combination with a laser focus that incorporates a point or line of zero intensity. In the past years, the concept has been transferred from microscopy to maskless laser lithography, namely direct laser writing (DLW), in order to overcome the diffraction limit for optical lithography. Herein, we propose and experimentally introduce a system that realizes such a molecular switch for lithography. Specifically, the population of intermediate-state photoenol isomers of α-methyl benzaldehydes generated by two-photon absorption at 700 nm fundamental wavelength can be reversibly depleted by simultaneous irradiation at 440 nm, suppressing the subsequent Diels-Alder cycloaddition reaction which constitutes the chemical core of the writing process. We demonstrate the potential of the proposed mechanism for STED-inspired DLW by covalently functionalizing the surface of glass substrates via the photoenol-driven STED-inspired process exploiting reversible photoenol activation with a polymerization initiator. Subsequently, macromolecules are grown from the functionalized areas and the spatially coded glass slides are characterized by atomic-force microscopy. Our approach allows lines with a full-width-at-half-maximum of down to 60 nm and line gratings with a lateral resolution of 100 nm to be written, both surpassing the diffraction limit.

  8. Two-color CW STED nanoscopy

    NASA Astrophysics Data System (ADS)

    Chen, Xuanze; Liu, Yujia; Yang, Xusan; Wang, Tingting; Alonas, Eric; Santangelo, Philip J.; Ren, Qiushi; Xi, Peng

    2013-02-01

    Fluorescent microscopy has become an essential tool to study biological molecules, pathways and events in living cells, tissues and animals. Meanwhile even the most advanced confocal microscopy can only yield optical resolution approaching Abbe diffraction limit of 200 nm. This is still larger than many subcellular structures, which are too small to be resolved in detail. These limitations have driven the development of super-resolution optical imaging methodologies over the past decade. In stimulated emission depletion (STED) microscopy, the excitation focus is overlapped by an intense doughnut-shaped spot to instantly de-excite markers from their fluorescent state to the ground state by stimulated emission. This effectively eliminates the periphery of the Point Spread Function (PSF), resulting in a narrower focal region, or super-resolution. Scanning a sharpened spot through the specimen renders images with sub-diffraction resolution. Multi-color STED imaging can present important structural and functional information for protein-protein interaction. In this work, we presented a two-color, synchronization-free STED microscopy with a Ti:Sapphire oscillator. The excitation wavelengths were 532nm and 635nm, respectively. With pump power of 4.6 W and sample irradiance of 310 mW, we achieved super-resolution as high as 71 nm. Human respiratory syncytial virus (hRSV) proteins were imaged with our two-color CW STED for co-localization analysis.

  9. Design of discrete and continuous super-resolving Toraldo pupils in the microwave range.

    PubMed

    Olmi, Luca; Bolli, Pietro; Mugnai, Daniela

    2018-03-20

    The concept of super-resolution refers to various methods for improving the angular resolution of an optical imaging system beyond the classical diffraction limit. In optical microscopy, several techniques have been successfully developed with the aim of narrowing the central lobe of the illumination point spread function. In astronomy, however, no similar techniques can be used. A feasible method to design antennas and telescopes with angular resolution better than the diffraction limit consists of using variable transmittance pupils. In particular, discrete binary phase masks (0 or π ) with finite phase-jump positions, known as Toraldo pupils (TPs), have the advantage of being easy to fabricate but offer relatively little flexibility in terms of achieving specific trade-offs between design parameters, such as the angular width of the main lobe and the intensity of sidelobes. In this paper, we show that a complex transmittance filter (equivalent to a continuous TP, i.e., consisting of infinitely narrow concentric rings) can achieve more easily the desired trade-off between design parameters. We also show how the super-resolution effect can be generated with both amplitude- and phase-only masks and confirm the expected performance with electromagnetic numerical simulations in the microwave range.

  10. Resolution limits of ultrafast ultrasound localization microscopy

    NASA Astrophysics Data System (ADS)

    Desailly, Yann; Pierre, Juliette; Couture, Olivier; Tanter, Mickael

    2015-11-01

    As in other imaging methods based on waves, the resolution of ultrasound imaging is limited by the wavelength. However, the diffraction-limit can be overcome by super-localizing single events from isolated sources. In recent years, we developed plane-wave ultrasound allowing frame rates up to 20 000 fps. Ultrafast processes such as rapid movement or disruption of ultrasound contrast agents (UCA) can thus be monitored, providing us with distinct punctual sources that could be localized beyond the diffraction limit. We previously showed experimentally that resolutions beyond λ/10 can be reached in ultrafast ultrasound localization microscopy (uULM) using a 128 transducer matrix in reception. Higher resolutions are theoretically achievable and the aim of this study is to predict the maximum resolution in uULM with respect to acquisition parameters (frequency, transducer geometry, sampling electronics). The accuracy of uULM is the error on the localization of a bubble, considered a point-source in a homogeneous medium. The proposed model consists in two steps: determining the timing accuracy of the microbubble echo in radiofrequency data, then transferring this time accuracy into spatial accuracy. The simplified model predicts a maximum resolution of 40 μm for a 1.75 MHz transducer matrix composed of two rows of 64 elements. Experimental confirmation of the model was performed by flowing microbubbles within a 60 μm microfluidic channel and localizing their blinking under ultrafast imaging (500 Hz frame rate). The experimental resolution, determined as the standard deviation in the positioning of the microbubbles, was predicted within 6 μm (13%) of the theoretical values and followed the analytical relationship with respect to the number of elements and depth. Understanding the underlying physical principles determining the resolution of superlocalization will allow the optimization of the imaging setup for each organ. Ultimately, accuracies better than the size of capillaries are achievable at several centimeter depths.

  11. FRIDA: diffraction-limited imaging and integral-field spectroscopy for the GTC

    NASA Astrophysics Data System (ADS)

    Watson, Alan M.; Acosta-Pulido, José A.; Álvarez-Núñez, Luis C.; Bringas-Rico, Vicente; Cardiel, Nicolás.; Cardona, Salvador; Chapa, Oscar; Díaz García, José Javier; Eikenberry, Stephen S.; Espejo, Carlos; Flores-Meza, Rubén. A.; Fuentes-Fernández, Jorge; Gallego, Jesús; Garcés Medina, José Leonardo; Garzón López, Francisco; Hammersley, Peter; Keiman, Carolina; Lara, Gerardo; López, José Alberto; López, Pablo L.; Lucero, Diana; Moreno Arce, Heidy; Pascual Ramirez, Sergio; Patrón Recio, Jesús; Prieto, Almudena; Rodríguez, Alberto José; Marco de la Rosa, José; Sánchez, Beatriz; Uribe, Jorge A.; Váldez Berriozabal, Francisco

    2016-08-01

    FRIDA is a diffraction-limited imager and integral-field spectrometer that is being built for the adaptive-optics focus of the Gran Telescopio Canarias. In imaging mode FRIDA will provide scales of 0.010, 0.020 and 0.040 arcsec/pixel and in IFS mode spectral resolutions of 1500, 4000 and 30,000. FRIDA is starting systems integration and is scheduled to complete fully integrated system tests at the laboratory by the end of 2017 and to be delivered to GTC shortly thereafter. In this contribution we present a summary of its design, fabrication, current status and potential scientific applications.

  12. Super-resolution atomic force photoactivated microscopy of biological samples (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Lee, Seunghyun; Kim, Hyemin; Shin, Seungjun; Doh, Junsang; Kim, Chulhong

    2017-03-01

    Optical microscopy (OM) and photoacoustic microscopy (PAM) have previously been used to image the optical absorption of intercellular features of biological cells. However, the optical diffraction limit ( 200 nm) makes it difficult for these modalities to image nanoscale inner cell structures and the distribution of internal cell components. Although super-resolution fluorescence microscopy, such as stimulated emission depletion microscopy (STED) and stochastic optical reconstruction microscopy (STORM), has successfully performed nanoscale biological imaging, these modalities require the use of exogenous fluorescence agents, which are unfavorable for biological samples. Our newly developed atomic force photoactivated microscopy (AFPM) can provide optical absorption images with nanoscale lateral resolution without any exogenous contrast agents. AFPM combines conventional atomic force microscopy (AFM) and an optical excitation system, and simultaneously provides multiple contrasts, such as the topography and magnitude of optical absorption. AFPM can detect the intrinsic optical absorption of samples with 8 nm lateral resolution, easily overcoming the diffraction limit. Using the label-free AFPM system, we have successfully imaged the optical absorption properties of a single melanoma cell (B16F10) and a rosette leaf epidermal cell of Arabidopsis (ecotype Columbia (Col-0)) with nanoscale lateral resolution. The remarkable images show the melanosome distribution of a melanoma cell and the biological structures of a plant cell. AFPM provides superior imaging of optical absorption with a nanoscale lateral resolution, and it promises to become widely used in biological and chemical research.

  13. Optical aperture synthesis with electronically connected telescopes

    PubMed Central

    Dravins, Dainis; Lagadec, Tiphaine; Nuñez, Paul D.

    2015-01-01

    Highest resolution imaging in astronomy is achieved by interferometry, connecting telescopes over increasingly longer distances and at successively shorter wavelengths. Here, we present the first diffraction-limited images in visual light, produced by an array of independent optical telescopes, connected electronically only, with no optical links between them. With an array of small telescopes, second-order optical coherence of the sources is measured through intensity interferometry over 180 baselines between pairs of telescopes, and two-dimensional images reconstructed. The technique aims at diffraction-limited optical aperture synthesis over kilometre-long baselines to reach resolutions showing details on stellar surfaces and perhaps even the silhouettes of transiting exoplanets. Intensity interferometry circumvents problems of atmospheric turbulence that constrain ordinary interferometry. Since the electronic signal can be copied, many baselines can be built up between dispersed telescopes, and over long distances. Using arrays of air Cherenkov telescopes, this should enable the optical equivalent of interferometric arrays currently operating at radio wavelengths. PMID:25880705

  14. Pushing the limits of spatial resolution with the Kuiper Airborne observatory

    NASA Technical Reports Server (NTRS)

    Lester, Daniel

    1994-01-01

    The study of astronomical objects at high spatial resolution in the far-IR is one of the most serious limitations to our work at these wavelengths, which carry information about the luminosity of dusty and obscured sources. At IR wavelengths shorter than 30 microns, ground based telescopes with large apertures at superb sites achieve diffraction-limited performance close to the seeing limit in the optical. At millimeter wavelengths, ground based interferometers achieve resolution that is close to this. The inaccessibility of the far-IR from the ground makes it difficult, however, to achieve complementary resolution in the far-IR. The 1983 IRAS survey, while extraordinarily sensitive, provides us with a sky map at a spatial resolution that is limited by detector size on a spatial scale that is far larger than that available in other wavelengths on the ground. The survey resolution is of order 4 min in the 100 micron bandpass, and 2 min at 60 microns (IRAS Explanatory Supplement, 1988). Information on a scale of 1' is available on some sources from the CPC. Deconvolution and image resolution using this database is one of the subjects of this workshop.

  15. Computational high-resolution optical imaging of the living human retina

    NASA Astrophysics Data System (ADS)

    Shemonski, Nathan D.; South, Fredrick A.; Liu, Yuan-Zhi; Adie, Steven G.; Scott Carney, P.; Boppart, Stephen A.

    2015-07-01

    High-resolution in vivo imaging is of great importance for the fields of biology and medicine. The introduction of hardware-based adaptive optics (HAO) has pushed the limits of optical imaging, enabling high-resolution near diffraction-limited imaging of previously unresolvable structures. In ophthalmology, when combined with optical coherence tomography, HAO has enabled a detailed three-dimensional visualization of photoreceptor distributions and individual nerve fibre bundles in the living human retina. However, the introduction of HAO hardware and supporting software adds considerable complexity and cost to an imaging system, limiting the number of researchers and medical professionals who could benefit from the technology. Here we demonstrate a fully automated computational approach that enables high-resolution in vivo ophthalmic imaging without the need for HAO. The results demonstrate that computational methods in coherent microscopy are applicable in highly dynamic living systems.

  16. Imaging fully hydrated whole cells by coherent x-ray diffraction microscopy.

    PubMed

    Nam, Daewoong; Park, Jaehyun; Gallagher-Jones, Marcus; Kim, Sangsoo; Kim, Sunam; Kohmura, Yoshiki; Naitow, Hisashi; Kunishima, Naoki; Yoshida, Takashi; Ishikawa, Tetsuya; Song, Changyong

    2013-03-01

    Nanoscale imaging of biological specimens in their native condition is of long-standing interest, in particular with direct, high resolution views of internal structures of intact specimens, though as yet progress has been limited. Here we introduce wet coherent x-ray diffraction microscopy capable of imaging fully hydrated and unstained biological specimens. Whole cell morphologies and internal structures better than 25 nm can be clearly visualized without contrast degradation.

  17. Bragg projection ptychography on niobium phase domains

    NASA Astrophysics Data System (ADS)

    Burdet, Nicolas; Shi, Xiaowen; Clark, Jesse N.; Huang, Xiaojing; Harder, Ross; Robinson, Ian

    2017-07-01

    Bragg projection ptychography (BPP) is a coherent x-ray diffraction imaging technique which combines the strengths of scanning microscopy with the phase contrast of x-ray ptychography. Here we apply it for high resolution imaging of the phase-shifted crystalline domains associated with epitaxial growth. The advantages of BPP are that the spatial extent of the sample is arbitrary, it is nondestructive, and it gives potentially diffraction limited spatial resolution. Here we demonstrate the application of BPP for revealing the domain structure caused by epitaxial misfit in a nanostructured metallic thin film. Experimental coherent diffraction data were collected from a niobium thin film, epitaxially grown on a sapphire substrate as the beam was scanned across the sample. The data were analyzed by BPP using a carefully selected combination of refinement procedures. The resulting image shows a close packed array of epitaxial domains, shifted with respect to each other due to misfit between the film and its substrate.

  18. SRRF: Universal live-cell super-resolution microscopy.

    PubMed

    Culley, Siân; Tosheva, Kalina L; Matos Pereira, Pedro; Henriques, Ricardo

    2018-08-01

    Super-resolution microscopy techniques break the diffraction limit of conventional optical microscopy to achieve resolutions approaching tens of nanometres. The major advantage of such techniques is that they provide resolutions close to those obtainable with electron microscopy while maintaining the benefits of light microscopy such as a wide palette of high specificity molecular labels, straightforward sample preparation and live-cell compatibility. Despite this, the application of super-resolution microscopy to dynamic, living samples has thus far been limited and often requires specialised, complex hardware. Here we demonstrate how a novel analytical approach, Super-Resolution Radial Fluctuations (SRRF), is able to make live-cell super-resolution microscopy accessible to a wider range of researchers. We show its applicability to live samples expressing GFP using commercial confocal as well as laser- and LED-based widefield microscopes, with the latter achieving long-term timelapse imaging with minimal photobleaching. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  19. Lensless transport-of-intensity phase microscopy and tomography with a color LED matrix

    NASA Astrophysics Data System (ADS)

    Zuo, Chao; Sun, Jiasong; Zhang, Jialin; Hu, Yan; Chen, Qian

    2015-07-01

    We demonstrate lens-less quantitative phase microscopy and diffraction tomography based on a compact on-chip platform, using only a CMOS image sensor and a programmable color LED array. Based on multi-wavelength transport-of- intensity phase retrieval and multi-angle illumination diffraction tomography, this platform offers high quality, depth resolved images with a lateral resolution of ˜3.7μm and an axial resolution of ˜5μm, over wide large imaging FOV of 24mm2. The resolution and FOV can be further improved by using a larger image sensors with small pixels straightforwardly. This compact, low-cost, robust, portable platform with a decent imaging performance may offer a cost-effective tool for telemedicine needs, or for reducing health care costs for point-of-care diagnostics in resource-limited environments.

  20. Angular reconstitution-based 3D reconstructions of nanomolecular structures from superresolution light-microscopy images

    PubMed Central

    Salas, Desirée; Le Gall, Antoine; Fiche, Jean-Bernard; Valeri, Alessandro; Ke, Yonggang; Bron, Patrick; Bellot, Gaetan

    2017-01-01

    Superresolution light microscopy allows the imaging of labeled supramolecular assemblies at a resolution surpassing the classical diffraction limit. A serious limitation of the superresolution approach is sample heterogeneity and the stochastic character of the labeling procedure. To increase the reproducibility and the resolution of the superresolution results, we apply multivariate statistical analysis methods and 3D reconstruction approaches originally developed for cryogenic electron microscopy of single particles. These methods allow for the reference-free 3D reconstruction of nanomolecular structures from two-dimensional superresolution projection images. Since these 2D projection images all show the structure in high-resolution directions of the optical microscope, the resulting 3D reconstructions have the best possible isotropic resolution in all directions. PMID:28811371

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

    PubMed

    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.

  2. Open data set of live cyanobacterial cells imaged using an X-ray laser

    NASA Astrophysics Data System (ADS)

    van der Schot, Gijs; Svenda, Martin; Maia, Filipe R. N. C.; Hantke, Max F.; Deponte, Daniel P.; Seibert, M. Marvin; Aquila, Andrew; Schulz, Joachim; Kirian, Richard A.; Liang, Mengning; Stellato, Francesco; Bari, Sadia; Iwan, Bianca; Andreasson, Jakob; Timneanu, Nicusor; Bielecki, Johan; Westphal, Daniel; Nunes de Almeida, Francisca; Odić, Duško; Hasse, Dirk; Carlsson, Gunilla H.; Larsson, Daniel S. D.; Barty, Anton; Martin, Andrew V.; Schorb, Sebastian; Bostedt, Christoph; Bozek, John D.; Carron, Sebastian; Ferguson, Ken; Rolles, Daniel; Rudenko, Artem; Epp, Sascha W.; Foucar, Lutz; Rudek, Benedikt; Erk, Benjamin; Hartmann, Robert; Kimmel, Nils; Holl, Peter; Englert, Lars; Loh, N. Duane; Chapman, Henry N.; Andersson, Inger; Hajdu, Janos; Ekeberg, Tomas

    2016-08-01

    Structural studies on living cells by conventional methods are limited to low resolution because radiation damage kills cells long before the necessary dose for high resolution can be delivered. X-ray free-electron lasers circumvent this problem by outrunning key damage processes with an ultra-short and extremely bright coherent X-ray pulse. Diffraction-before-destruction experiments provide high-resolution data from cells that are alive when the femtosecond X-ray pulse traverses the sample. This paper presents two data sets from micron-sized cyanobacteria obtained at the Linac Coherent Light Source, containing a total of 199,000 diffraction patterns. Utilizing this type of diffraction data will require the development of new analysis methods and algorithms for studying structure and structural variability in large populations of cells and to create abstract models. Such studies will allow us to understand living cells and populations of cells in new ways. New X-ray lasers, like the European XFEL, will produce billions of pulses per day, and could open new areas in structural sciences.

  3. Open data set of live cyanobacterial cells imaged using an X-ray laser.

    PubMed

    van der Schot, Gijs; Svenda, Martin; Maia, Filipe R N C; Hantke, Max F; DePonte, Daniel P; Seibert, M Marvin; Aquila, Andrew; Schulz, Joachim; Kirian, Richard A; Liang, Mengning; Stellato, Francesco; Bari, Sadia; Iwan, Bianca; Andreasson, Jakob; Timneanu, Nicusor; Bielecki, Johan; Westphal, Daniel; Nunes de Almeida, Francisca; Odić, Duško; Hasse, Dirk; Carlsson, Gunilla H; Larsson, Daniel S D; Barty, Anton; Martin, Andrew V; Schorb, Sebastian; Bostedt, Christoph; Bozek, John D; Carron, Sebastian; Ferguson, Ken; Rolles, Daniel; Rudenko, Artem; Epp, Sascha W; Foucar, Lutz; Rudek, Benedikt; Erk, Benjamin; Hartmann, Robert; Kimmel, Nils; Holl, Peter; Englert, Lars; Loh, N Duane; Chapman, Henry N; Andersson, Inger; Hajdu, Janos; Ekeberg, Tomas

    2016-08-01

    Structural studies on living cells by conventional methods are limited to low resolution because radiation damage kills cells long before the necessary dose for high resolution can be delivered. X-ray free-electron lasers circumvent this problem by outrunning key damage processes with an ultra-short and extremely bright coherent X-ray pulse. Diffraction-before-destruction experiments provide high-resolution data from cells that are alive when the femtosecond X-ray pulse traverses the sample. This paper presents two data sets from micron-sized cyanobacteria obtained at the Linac Coherent Light Source, containing a total of 199,000 diffraction patterns. Utilizing this type of diffraction data will require the development of new analysis methods and algorithms for studying structure and structural variability in large populations of cells and to create abstract models. Such studies will allow us to understand living cells and populations of cells in new ways. New X-ray lasers, like the European XFEL, will produce billions of pulses per day, and could open new areas in structural sciences.

  4. Open data set of live cyanobacterial cells imaged using an X-ray laser

    PubMed Central

    van der Schot, Gijs; Svenda, Martin; Maia, Filipe R.N.C.; Hantke, Max F.; DePonte, Daniel P.; Seibert, M. Marvin; Aquila, Andrew; Schulz, Joachim; Kirian, Richard A.; Liang, Mengning; Stellato, Francesco; Bari, Sadia; Iwan, Bianca; Andreasson, Jakob; Timneanu, Nicusor; Bielecki, Johan; Westphal, Daniel; Nunes de Almeida, Francisca; Odić, Duško; Hasse, Dirk; Carlsson, Gunilla H.; Larsson, Daniel S.D.; Barty, Anton; Martin, Andrew V.; Schorb, Sebastian; Bostedt, Christoph; Bozek, John D.; Carron, Sebastian; Ferguson, Ken; Rolles, Daniel; Rudenko, Artem; Epp, Sascha W.; Foucar, Lutz; Rudek, Benedikt; Erk, Benjamin; Hartmann, Robert; Kimmel, Nils; Holl, Peter; Englert, Lars; Loh, N. Duane; Chapman, Henry N.; Andersson, Inger; Hajdu, Janos; Ekeberg, Tomas

    2016-01-01

    Structural studies on living cells by conventional methods are limited to low resolution because radiation damage kills cells long before the necessary dose for high resolution can be delivered. X-ray free-electron lasers circumvent this problem by outrunning key damage processes with an ultra-short and extremely bright coherent X-ray pulse. Diffraction-before-destruction experiments provide high-resolution data from cells that are alive when the femtosecond X-ray pulse traverses the sample. This paper presents two data sets from micron-sized cyanobacteria obtained at the Linac Coherent Light Source, containing a total of 199,000 diffraction patterns. Utilizing this type of diffraction data will require the development of new analysis methods and algorithms for studying structure and structural variability in large populations of cells and to create abstract models. Such studies will allow us to understand living cells and populations of cells in new ways. New X-ray lasers, like the European XFEL, will produce billions of pulses per day, and could open new areas in structural sciences. PMID:27479514

  5. Opto-mechanical design and development of a 460mm diffractive transmissive telescope

    NASA Astrophysics Data System (ADS)

    Qi, Bo; Wang, Lihua; Cui, Zhangang; Bian, Jiang; Xiang, Sihua; Ma, Haotong; Fan, Bin

    2018-01-01

    Using lightweight, replicated diffractive optics, we can construct extremely large aperture telescopes in space.The transmissive primary significantly reduces the sensitivities to out of plane motion as compared to reflective systems while reducing the manufacturing time and costs. This paper focuses on the design, fabrication and ground demonstration of a 460mm diffractive transmissive telescope the primary F/# is 6, optical field of view is 0.2° imagine bandwidth is 486nm 656nm.The design method of diffractive optical system was verified, the ability to capture a high-quality image using diffractive telescope collection optics was tested.The results show that the limit resolution is 94lp/mm, the diffractive system has a good imagine performance with broad bandwidths. This technology is particularly promising as a means to achieve extremely large optical primaries from compact, lightweight packages.

  6. Restoration of multichannel microwave radiometric images

    NASA Technical Reports Server (NTRS)

    Chin, R. T.; Yeh, C. L.; Olson, W. S.

    1983-01-01

    A constrained iterative image restoration method is applied to multichannel diffraction-limited imagery. This method is based on the Gerchberg-Papoulis algorithm utilizing incomplete information and partial constraints. The procedure is described using the orthogonal projection operators which project onto two prescribed subspaces iteratively. Some of its properties and limitations are also presented. The selection of appropriate constraints was emphasized in a practical application. Multichannel microwave images, each having different spatial resolution, were restored to a common highest resolution to demonstrate the effectiveness of the method. Both noise-free and noisy images were used in this investigation.

  7. MIPS - The Multiband Imaging Photometer for SIRTF. [Multiband Imaging Photometer for SIRTF

    NASA Technical Reports Server (NTRS)

    Rieke, G. H.; Lada, C.; Lebofsky, M.; Low, F.; Strittmatter, P.; Young, E.; Arens, J.; Beichman, C.; Gautier, T. N.; Werner, M.

    1986-01-01

    The Multiband Imaging Photometer for SIRTF (MIPS) is to be designed to reach as closely as possible the fundamental sensitivity and angular resolution limits for SIRTF over the 3 to 700 micron spectral region. It will use high performance photoconductive detectors from 3 to 200 micron with integrating JFET amplifiers. From 200 to 700 microns, the MIPS will use a bolometer cooled by an adiabatic demagnetization refrigerator. Over much of its operating range, the MIPS will make possible observations at and beyond the conventional Rayleigh diffraction limit of angular resolution.

  8. MIPS - The Multiband Imaging Photometer for SIRTF

    NASA Technical Reports Server (NTRS)

    Rieke, G. H.; Lada, C.; Lebofsky, M.; Low, F.; Strittmatter, P.; Young, E.; Beichman, C.; Gautier, T. N.; Mould, J.; Werner, M.

    1986-01-01

    The Multiband Imaging Photometer System (MIPS) for SIRTF is to be designed to reach as closely as possible the fundamental sensitivity and angular resolution limits for SIRTF over the 3 to 700 microns spectral region. It will use high performance photoconductive detectors from 3 to 200 microns with integrating JFET amplifiers. From 200 to 700 microns, the MIPS will use a bolometer cooled by an adiabatic demagnetization refrigerator. Over much of its operating range, the MIPS will make possible observations at and beyond the conventional Rayleigh diffraction limit of angular resolution.

  9. Feasibility of detecting near-surface feature with Rayleigh-wave diffraction

    USGS Publications Warehouse

    Xia, J.; Nyquist, Jonathan E.; Xu, Y.; Roth, M.J.S.; Miller, R.D.

    2007-01-01

    Detection of near-surfaces features such as voids and faults is challenging due to the complexity of near-surface materials and the limited resolution of geophysical methods. Although multichannel, high-frequency, surface-wave techniques can provide reliable shear (S)-wave velocities in different geological settings, they are not suitable for detecting voids directly based on anomalies of the S-wave velocity because of limitations on the resolution of S-wave velocity profiles inverted from surface-wave phase velocities. Therefore, we studied the feasibility of directly detecting near-surfaces features with surface-wave diffractions. Based on the properties of surface waves, we have derived a Rayleigh-wave diffraction traveltime equation. We also have solved the equation for the depth to the top of a void and an average velocity of Rayleigh waves. Using these equations, the depth to the top of a void/fault can be determined based on traveltime data from a diffraction curve. In practice, only two diffraction times are necessary to define the depth to the top of a void/fault and the average Rayleigh-wave velocity that generates the diffraction curve. We used four two-dimensional square voids to demonstrate the feasibility of detecting a void with Rayleigh-wave diffractions: a 2??m by 2??m with a depth to the top of the void of 2??m, 4??m by 4??m with a depth to the top of the void of 7??m, and 6??m by 6??m with depths to the top of the void 12??m and 17??m. We also modeled surface waves due to a vertical fault. Rayleigh-wave diffractions were recognizable for all these models after FK filtering was applied to the synthetic data. The Rayleigh-wave diffraction traveltime equation was verified by the modeled data. Modeling results suggested that FK filtering is critical to enhance diffracted surface waves. A real-world example is presented to show how to utilize the derived equation of surface-wave diffractions. ?? 2006 Elsevier B.V. All rights reserved.

  10. Conical diffraction as a versatile building block to implement new imaging modalities for superresolution in fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Fallet, Clément; Caron, Julien; Oddos, Stephane; Tinevez, Jean-Yves; Moisan, Lionel; Sirat, Gabriel Y.; Braitbart, Philippe O.; Shorte, Spencer L.

    2014-08-01

    We present a new technology for super-resolution fluorescence imaging, based on conical diffraction. Conical diffraction is a linear, singular phenomenon taking place when a polarized beam is diffracted through a biaxial crystal. The illumination patterns generated by conical diffraction are more compact than the classical Gaussian beam; we use them to generate a super-resolution imaging modality. Conical Diffraction Microscopy (CODIM) resolution enhancement can be achieved with any type of objective on any kind of sample preparation and standard fluorophores. Conical diffraction can be used in multiple fashion to create new and disruptive technologies for super-resolution microscopy. This paper will focus on the first one that has been implemented and give a glimpse at what the future of microscopy using conical diffraction could be.

  11. Improving the diffraction of apoA-IV crystals through extreme dehydration.

    PubMed

    Deng, Xiaodi; Davidson, W Sean; Thompson, Thomas B

    2012-01-01

    Apolipoproteins are the protein component of high-density lipoproteins (HDL), which are necessary for mobilizing lipid-like molecules throughout the body. Apolipoproteins undergo self-association, especially at higher concentrations, making them difficult to crystallize. Here, the crystallization and diffraction of the core fragment of apolipoprotein A-IV (apoA-IV), consisting of residues 64-335, is presented. ApoA-IV(64-335) crystallized readily in a variety of hexagonal (P6) morphologies with similar unit-cell parameters, all containing a long axis of nearly 550 Å in length. Preliminary diffraction experiments with the different crystal morphologies all resulted in limited streaky diffraction to 3.5 Å resolution. Crystal dehydration was applied to the different morphologies with variable success and was also used as a quality indicator of crystal-growth conditions. The results show that the morphologies that withstood the most extreme dehydration conditions showed the greatest improvement in diffraction. One morphology in particular was able to withstand dehydration in 60% PEG 3350 for over 12 h, which resulted in well defined intensities to 2.7 Å resolution. These results suggest that the approach of integrating dehydration with variation in crystal-growth conditions might be a general technique to optimize diffraction. © 2012 International Union of Crystallography. All rights reserved.

  12. High throughput screening of ligand binding to macromolecules using high resolution powder diffraction

    DOEpatents

    Von Dreele, Robert B.; D'Amico, Kevin

    2006-10-31

    A process is provided for the high throughput screening of binding of ligands to macromolecules using high resolution powder diffraction data including producing a first sample slurry of a selected polycrystalline macromolecule material and a solvent, producing a second sample slurry of a selected polycrystalline macromolecule material, one or more ligands and the solvent, obtaining a high resolution powder diffraction pattern on each of said first sample slurry and the second sample slurry, and, comparing the high resolution powder diffraction pattern of the first sample slurry and the high resolution powder diffraction pattern of the second sample slurry whereby a difference in the high resolution powder diffraction patterns of the first sample slurry and the second sample slurry provides a positive indication for the formation of a complex between the selected polycrystalline macromolecule material and at least one of the one or more ligands.

  13. Throughput of diffraction-limited field optics systems for infrared and millimetric telescopes

    NASA Technical Reports Server (NTRS)

    Hildebrand, R. H.; Winston, R.

    1982-01-01

    Telescopes for submillimeter wavelengths have point spread functions some millimeters or centimeters in diameter, but the detectors may be only fractions of a millimeter in size. Thus a field aperture and collecting optics are needed. Optimizing the aperture by a calculation of the effects of diffraction on signal and resolution as a function of size of the collecting aperture is shown. Calculations are compared to experimental results from observations of Mars at submillimeter wavelengths.

  14. Enhancing resolution and contrast in second-harmonic generation microscopy using an advanced maximum likelihood estimation restoration method

    NASA Astrophysics Data System (ADS)

    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.

  15. An assessment of the resolution limitation due to radiation-damage in X-ray diffraction microscopy

    DOE PAGES

    Howells, M. R.; Beetz, T.; Chapman, H. N.; ...

    2008-11-17

    X-ray diffraction microscopy (XDM) is a new form of x-ray imaging that is being practiced at several third-generation synchrotron-radiation x-ray facilities. Nine years have elapsed since the technique was first introduced and it has made rapid progress in demonstrating high-resolution three-dimensional imaging and promises few-nm resolution with much larger samples than can be imaged in the transmission electron microscope. Both life- and materials-science applications of XDM are intended, and it is expected that the principal limitation to resolution will be radiation damage for life science and the coherent power of available x-ray sources for material science. In this paper wemore » address the question of the role of radiation damage. We use a statistical analysis based on the so-called "dose fractionation theorem" of Hegerl and Hoppe to calculate the dose needed to make an image of a single life-science sample by XDM with a given resolution. We find that for simply-shaped objects the needed dose scales with the inverse fourth power of the resolution and present experimental evidence to support this finding. To determine the maximum tolerable dose we have assembled a number of data taken from the literature plus some measurements of our own which cover ranges of resolution that are not well covered otherwise. The conclusion of this study is that, based on the natural contrast between protein and water and "Rose-criterion" image quality, one should be able to image a frozen-hydrated biological sample using XDM at a resolution of about 10 nm.« less

  16. Optical method for high magnification imaging and video recording of live cells at sub-micron resolution

    NASA Astrophysics Data System (ADS)

    Romo, Jaime E., Jr.

    Optical microscopy, the most common technique for viewing living microorganisms, is limited in resolution by Abbe's criterion. Recent microscopy techniques focus on circumnavigating the light diffraction limit by using different methods to obtain the topography of the sample. Systems like the AFM and SEM provide images with fields of view in the nanometer range with high resolvable detail, however these techniques are expensive, and limited in their ability to document live cells. The Dino-Lite digital microscope coupled with the Zeiss Axiovert 25 CFL microscope delivers a cost-effective method for recording live cells. Fields of view ranging from 8 microns to 300 microns with fair resolution provide a reliable method for discovering native cell structures at the nanoscale. In this report, cultured HeLa cells are recorded using different optical configurations resulting in documentation of cell dynamics at high magnification and resolution.

  17. Limitation of Liquid Crystal on Silicon Spatial Light Modular for Holographic Three-dimensional Displays

    NASA Technical Reports Server (NTRS)

    Wang, Xinghua; Wang, Bin; Bos, Philip J.; Anderson, James E.; Kujawinska, Malgorzata; Pouch, John; Miranda, Feliz

    2004-01-01

    In a 3-D display system based on an opto-electronic reconstruction of a digitally recorded hologram, the field of view of such a system is limited by the spatial resolution of the liquid crystal on silicon (LCOS) spatial light modular (SLM) used to perform the opto-electronic reconstruction. In this article, the special resolution limitation of LCOS SLM associated with the fringe field effect and interpixel coupling is determined by the liquid crystal detector simulation and the Finite Difference Time Domain (FDTD) simulation. The diffraction efficiency loss associated with the imperfection in the phase profile is studied with an example of opto-electronic reconstruction of an amplitude object. A high spatial resolution LCOS SLM with a wide reconstruction angle is proposed.

  18. Homology‐based hydrogen bond information improves crystallographic structures in the PDB

    PubMed Central

    van Beusekom, Bart; Touw, Wouter G.; Tatineni, Mahidhar; Somani, Sandeep; Rajagopal, Gunaretnam; Luo, Jinquan; Gilliland, Gary L.; Perrakis, Anastassis

    2017-01-01

    Abstract The Protein Data Bank (PDB) is the global archive for structural information on macromolecules, and a popular resource for researchers, teachers, and students, amassing more than one million unique users each year. Crystallographic structure models in the PDB (more than 100,000 entries) are optimized against the crystal diffraction data and geometrical restraints. This process of crystallographic refinement typically ignored hydrogen bond (H‐bond) distances as a source of information. However, H‐bond restraints can improve structures at low resolution where diffraction data are limited. To improve low‐resolution structure refinement, we present methods for deriving H‐bond information either globally from well‐refined high‐resolution structures from the PDB‐REDO databank, or specifically from on‐the‐fly constructed sets of homologous high‐resolution structures. Refinement incorporating HOmology DErived Restraints (HODER), improves geometrical quality and the fit to the diffraction data for many low‐resolution structures. To make these improvements readily available to the general public, we applied our new algorithms to all crystallographic structures in the PDB: using massively parallel computing, we constructed a new instance of the PDB‐REDO databank (https://pdb-redo.eu). This resource is useful for researchers to gain insight on individual structures, on specific protein families (as we demonstrate with examples), and on general features of protein structure using data mining approaches on a uniformly treated dataset. PMID:29168245

  19. Homology-based hydrogen bond information improves crystallographic structures in the PDB.

    PubMed

    van Beusekom, Bart; Touw, Wouter G; Tatineni, Mahidhar; Somani, Sandeep; Rajagopal, Gunaretnam; Luo, Jinquan; Gilliland, Gary L; Perrakis, Anastassis; Joosten, Robbie P

    2018-03-01

    The Protein Data Bank (PDB) is the global archive for structural information on macromolecules, and a popular resource for researchers, teachers, and students, amassing more than one million unique users each year. Crystallographic structure models in the PDB (more than 100,000 entries) are optimized against the crystal diffraction data and geometrical restraints. This process of crystallographic refinement typically ignored hydrogen bond (H-bond) distances as a source of information. However, H-bond restraints can improve structures at low resolution where diffraction data are limited. To improve low-resolution structure refinement, we present methods for deriving H-bond information either globally from well-refined high-resolution structures from the PDB-REDO databank, or specifically from on-the-fly constructed sets of homologous high-resolution structures. Refinement incorporating HOmology DErived Restraints (HODER), improves geometrical quality and the fit to the diffraction data for many low-resolution structures. To make these improvements readily available to the general public, we applied our new algorithms to all crystallographic structures in the PDB: using massively parallel computing, we constructed a new instance of the PDB-REDO databank (https://pdb-redo.eu). This resource is useful for researchers to gain insight on individual structures, on specific protein families (as we demonstrate with examples), and on general features of protein structure using data mining approaches on a uniformly treated dataset. © 2017 The Protein Society.

  20. Synchrotron FTIR Imaging For The Identification Of Cell Types Within Human Tissues

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Walsh, Michael J.; Pounder, F. Nell; Nasse, Michael J.

    2010-02-03

    The use of synchrotron Fourier Transform Infrared spectroscopy (S-FTIR) has been shown to be a very promising tool for biomedical research. S-FTIR spectroscopy allows for the fast acquisition of infrared (IR) spectra at a spatial resolution approaching the IR diffraction limit. The development of the Infrared Environmental Imaging (IRENI) beamline at the Synchrotron Radiation Center (SRC) at the University of Wisconsin-Madison has allowed for diffraction limited imaging measurements of cells in human prostate and breast tissues. This has allowed for the identification of cell types within tissues that would otherwise not have been resolvable using conventional FTIR sources.

  1. White-light diffraction phase microscopy at doubled space-bandwidth product.

    PubMed

    Shan, Mingguang; Kandel, Mikhail E; Majeed, Hassaan; Nastasa, Viorel; Popescu, Gabriel

    2016-12-12

    White light diffraction microscopy (wDPM) is a quantitative phase imaging method that benefits from both temporal and spatial phase sensitivity, granted, respectively, by the common-path geometry and white light illumination. However, like all off-axis quantitative phase imaging methods, wDPM is characterized by a reduced space-bandwidth product compared to phase shifting approaches. This happens essentially because the ultimate resolution of the image is governed by the period of the interferogram and not just the diffraction limit. As a result, off-axis techniques generates single-shot, i.e., high time-bandwidth, phase measurements, at the expense of either spatial resolution or field of view. Here, we show that combining phase-shifting and off-axis, the original space-bandwidth is preserved. Specifically, we developed phase-shifting diffraction phase microscopy with white light, in which we measure and combine two phase shifted interferograms. Due to the white light illumination, the phase images are characterized by low spatial noise, i.e., <1nm pathlength. We illustrate the operation of the instrument with test samples, blood cells, and unlabeled prostate tissue biopsy.

  2. Comparative Analysis of Thaumatin Crystals Grown on Earth and in Microgravity. Experiment 23

    NASA Technical Reports Server (NTRS)

    Ng, Joseph D.; Lorber, Bernard; Giege, Richard; Koszelak, Stanley; Day, John; Greenwood, Aaron; McPherson, Alexander

    1998-01-01

    The protein thaumatin was studied as a model macromolecule for crystallization in microgravity environment experiments conducted on two U.S. Space Shuttle missions (second United States Microgravity Laboratory (USML-2) and Life and Microgravity Spacelab (LMS)). In this investigation we evaluated and compared the quality of space- and Earth-grown thaumatin crystals using x-ray diffraction analysis and characterized them according to crystal size, diffraction resolution limit, and mosaicity. Two different approaches for growing thaumatin crystals in the microgravity environment, dialysis and liquid-liquid diffusion, were employed as a joint experiment by our two investigative teams. Thaumatin crystals grown under a microgravity environment were generally larger in volume with fewer total crystals. They diffracted to significantly higher resolution and with improved diffraction properties as judged by relative Wilson plots. The mosaicity for space-grown crystals was significantly less than for those grown on Earth. Increasing concentrations of protein in the crystallization chambers under microgravity lead to larger crystals. The data presented here lend further support to the idea that protein crystals of improved quality can be obtained in a microgravity environment.

  3. Measuring the spatial resolution of an optical system in an undergraduate optics laboratory

    NASA Astrophysics Data System (ADS)

    Leung, Calvin; Donnelly, T. D.

    2017-06-01

    Two methods of quantifying the spatial resolution of a camera are described, performed, and compared, with the objective of designing an imaging-system experiment for students in an undergraduate optics laboratory. With the goal of characterizing the resolution of a typical digital single-lens reflex (DSLR) camera, we motivate, introduce, and show agreement between traditional test-target contrast measurements and the technique of using Fourier analysis to obtain the modulation transfer function (MTF). The advantages and drawbacks of each method are compared. Finally, we explore the rich optical physics at work in the camera system by calculating the MTF as a function of wavelength and f-number. For example, we find that the Canon 40D demonstrates better spatial resolution at short wavelengths, in accordance with scalar diffraction theory, but is not diffraction-limited, being significantly affected by spherical aberration. The experiment and data analysis routines described here can be built and written in an undergraduate optics lab setting.

  4. Characterization of Ultra-fine Grained and Nanocrystalline Materials Using Transmission Kikuchi Diffraction

    PubMed Central

    Proust, Gwénaëlle; Trimby, Patrick; Piazolo, Sandra; Retraint, Delphine

    2017-01-01

    One of the challenges in microstructure analysis nowadays resides in the reliable and accurate characterization of ultra-fine grained (UFG) and nanocrystalline materials. The traditional techniques associated with scanning electron microscopy (SEM), such as electron backscatter diffraction (EBSD), do not possess the required spatial resolution due to the large interaction volume between the electrons from the beam and the atoms of the material. Transmission electron microscopy (TEM) has the required spatial resolution. However, due to a lack of automation in the analysis system, the rate of data acquisition is slow which limits the area of the specimen that can be characterized. This paper presents a new characterization technique, Transmission Kikuchi Diffraction (TKD), which enables the analysis of the microstructure of UFG and nanocrystalline materials using an SEM equipped with a standard EBSD system. The spatial resolution of this technique can reach 2 nm. This technique can be applied to a large range of materials that would be difficult to analyze using traditional EBSD. After presenting the experimental set up and describing the different steps necessary to realize a TKD analysis, examples of its use on metal alloys and minerals are shown to illustrate the resolution of the technique and its flexibility in term of material to be characterized. PMID:28447998

  5. Characterization of Ultra-fine Grained and Nanocrystalline Materials Using Transmission Kikuchi Diffraction.

    PubMed

    Proust, Gwénaëlle; Trimby, Patrick; Piazolo, Sandra; Retraint, Delphine

    2017-04-01

    One of the challenges in microstructure analysis nowadays resides in the reliable and accurate characterization of ultra-fine grained (UFG) and nanocrystalline materials. The traditional techniques associated with scanning electron microscopy (SEM), such as electron backscatter diffraction (EBSD), do not possess the required spatial resolution due to the large interaction volume between the electrons from the beam and the atoms of the material. Transmission electron microscopy (TEM) has the required spatial resolution. However, due to a lack of automation in the analysis system, the rate of data acquisition is slow which limits the area of the specimen that can be characterized. This paper presents a new characterization technique, Transmission Kikuchi Diffraction (TKD), which enables the analysis of the microstructure of UFG and nanocrystalline materials using an SEM equipped with a standard EBSD system. The spatial resolution of this technique can reach 2 nm. This technique can be applied to a large range of materials that would be difficult to analyze using traditional EBSD. After presenting the experimental set up and describing the different steps necessary to realize a TKD analysis, examples of its use on metal alloys and minerals are shown to illustrate the resolution of the technique and its flexibility in term of material to be characterized.

  6. What is the diffraction limit? From Airy to Abbe using direct numerical integration

    NASA Astrophysics Data System (ADS)

    Calm, Y. M.; Merlo, J. M.; Burns, M. J.; Kempa, K.; Naughton, M. J.

    The resolution of a conventional optical microscope is sometimes taken from Airy's point spread function (PSF), 0 . 61 λ / NA , and sometimes from Abbe, λ / 2 NA , where NA is the numerical aperture, however modern fluorescence and near-field optical microscopies achieve spatial resolution far better than either of these limits. There is a new category of 2D metamaterials called planar optical elements (POEs), which have a microscopic thickness (< λ), macroscopic transverse dimensions (> 100 λ), and are composed of an array of nanostructured light scatterers. POEs are found in a range of micro- and nano-photonic technologies, and will influence the future optical nanoscopy. With this pretext, we shed some light on the 'diffraction limit' by numerically evaluating Kirchhoff's scalar formulae (in their exact form) and identifying the features of highly non-paraxial, 3D PSFs. We show that the Airy and Abbe criteria are connected, and we comment on the design rules for a particular type of POE: the flat lens. This work is supported by the W. M. Keck Foundation.

  7. A pulse-front-tilt–compensated streaked optical spectrometer with high throughput and picosecond time resolution

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Katz, J., E-mail: jkat@lle.rochester.edu; Boni, R.; Rivlis, R.

    A high-throughput, broadband optical spectrometer coupled to the Rochester optical streak system equipped with a Photonis P820 streak tube was designed to record time-resolved spectra with 1-ps time resolution. Spectral resolution of 0.8 nm is achieved over a wavelength coverage range of 480 to 580 nm, using a 300-groove/mm diffraction grating in conjunction with a pair of 225-mm-focal-length doublets operating at an f/2.9 aperture. Overall pulse-front tilt across the beam diameter generated by the diffraction grating is reduced by preferentially delaying discrete segments of the collimated input beam using a 34-element reflective echelon optic. The introduced delay temporally aligns themore » beam segments and the net pulse-front tilt is limited to the accumulation across an individual sub-element. The resulting spectrometer design balances resolving power and pulse-front tilt while maintaining high throughput.« less

  8. Macromolecule Crystal Quality Improvement in Microgravity: The Role of Impurities

    NASA Technical Reports Server (NTRS)

    Judge, Russell A.; Snell, Edward H.; Pusey, Marc L.; Sportiello, Michael G.; Todd, Paul; Bellamy, Henry; Borgstahl, Gloria E.; Pokros, Matt; Cassanto, John M.

    2000-01-01

    While macromolecule impurities may affect crystal size and morphology the over-riding question is; "How do macromolecule impurities effect crystal X-ray quality and diffraction resolution?" In the case of chicken egg white lysozyme, crystals can be grown in the presence of a number of impurities without affecting diffraction resolution. One impurity however, the lysozyme dimer, does negatively impact the X-ray crystal properties. Crystal quality improvement as a result of better partitioning of this impurity during crystallization in microgravity has been reported'. In our recent experimental work dimer partitioning was found to be not significantly different between the two environments. Mosaicity analysis of pure crystals showed a reduced mosaicity and increased signal to noise for the microgravity grown crystals. Dimer incorporation however, did greatly reduce the resolution limit in both ground and microgravity grown crystals. These results indicate that impurity effects in microgravity are complex and may rely on the conditions or techniques employed.

  9. Femtosecond X-ray coherent diffraction of aligned amyloid fibrils on low background graphene.

    PubMed

    Seuring, Carolin; Ayyer, Kartik; Filippaki, Eleftheria; Barthelmess, Miriam; Longchamp, Jean-Nicolas; Ringler, Philippe; Pardini, Tommaso; Wojtas, David H; Coleman, Matthew A; Dörner, Katerina; Fuglerud, Silje; Hammarin, Greger; Habenstein, Birgit; Langkilde, Annette E; Loquet, Antoine; Meents, Alke; Riek, Roland; Stahlberg, Henning; Boutet, Sébastien; Hunter, Mark S; Koglin, Jason; Liang, Mengning; Ginn, Helen M; Millane, Rick P; Frank, Matthias; Barty, Anton; Chapman, Henry N

    2018-05-09

    Here we present a new approach to diffraction imaging of amyloid fibrils, combining a free-standing graphene support and single nanofocused X-ray pulses of femtosecond duration from an X-ray free-electron laser. Due to the very low background scattering from the graphene support and mutual alignment of filaments, diffraction from tobacco mosaic virus (TMV) filaments and amyloid protofibrils is obtained to 2.7 Å and 2.4 Å resolution in single diffraction patterns, respectively. Some TMV diffraction patterns exhibit asymmetry that indicates the presence of a limited number of axial rotations in the XFEL focus. Signal-to-noise levels from individual diffraction patterns are enhanced using computational alignment and merging, giving patterns that are superior to those obtainable from synchrotron radiation sources. We anticipate that our approach will be a starting point for further investigations into unsolved structures of filaments and other weakly scattering objects.

  10. Birefringent coherent diffraction imaging

    NASA Astrophysics Data System (ADS)

    Karpov, Dmitry; dos Santos Rolo, Tomy; Rich, Hannah; Kryuchkov, Yuriy; Kiefer, Boris; Fohtung, E.

    2016-10-01

    Directional dependence of the index of refraction contains a wealth of information about anisotropic optical properties in semiconducting and insulating materials. Here we present a novel high-resolution lens-less technique that uses birefringence as a contrast mechanism to map the index of refraction and dielectric permittivity in optically anisotropic materials. We applied this approach successfully to a liquid crystal polymer film using polarized light from helium neon laser. This approach is scalable to imaging with diffraction-limited resolution, a prospect rapidly becoming a reality in view of emergent brilliant X-ray sources. Applications of this novel imaging technique are in disruptive technologies, including novel electronic devices, in which both charge and spin carry information as in multiferroic materials and photonic materials such as light modulators and optical storage.

  11. Label-free super-resolution with coherent nonlinear structured-illumination microscopy

    NASA Astrophysics Data System (ADS)

    Huttunen, Mikko J.; Abbas, Aazad; Upham, Jeremy; Boyd, Robert W.

    2017-08-01

    Structured-illumination microscopy enables up to a two-fold lateral resolution improvement by spatially modulating the intensity profile of the illumination beam. We propose a novel way to generalize the concept of structured illumination to nonlinear widefield modalities by spatially modulating, instead of field intensities, the phase of the incident field while interferometrically measuring the complex-valued scattered field. We numerically demonstrate that for second-order and third-order processes an almost four- and six-fold increase in lateral resolution is achievable, respectively. This procedure overcomes the conventional Abbe diffraction limit and provides new possibilities for label-free super-resolution microscopy.

  12. Spatial resolution limitation of liquid crystal spatial light modulator

    NASA Astrophysics Data System (ADS)

    Wang, Xinghua; Wang, Bin; McManamon, Paul F., III; Pouch, John J.; Miranda, Felix A.; Anderson, James E.; Bos, Philip J.

    2004-10-01

    The effect of fringing electric fields in a liquid crystal (LC) Optical Phased Array (OPA), also referred to as a spatial light modulator (SLM), is a governing factor that determines the diffraction efficiency (DE) of the LC OPA for high resolution spatial phase modulation. In this article, the fringing field effect in a high resolution LC OPA is studied by accurate modeling the DE of the LC blazed gratings by LC director simulation and Finite Difference Time Domain (FDTD) simulation. Influence factors that contribute significantly to the DE are discussed. Such results provide fundamental understanding for high resolution LC devices.

  13. Overcoming turbulence-induced space-variant blur by using phase-diverse speckle.

    PubMed

    Thelen, Brian J; Paxman, Richard G; Carrara, David A; Seldin, John H

    2009-01-01

    Space-variant blur occurs when imaging through volume turbulence over sufficiently large fields of view. Space-variant effects are particularly severe in horizontal-path imaging, slant-path (air-to-ground or ground-to-air) geometries, and ground-based imaging of low-elevation satellites or astronomical objects. In these geometries, the isoplanatic angle can be comparable to or even smaller than the diffraction-limited resolution angle. We report on a postdetection correction method that seeks to correct for the effects of space-variant aberrations, with the goal of reconstructing near-diffraction-limited imagery. Our approach has been to generalize the method of phase-diverse speckle (PDS) by using a physically motivated distributed-phase-screen model. Simulation results are presented that demonstrate the reconstruction of near-diffraction-limited imagery under both matched and mismatched model assumptions. In addition, we present evidence that PDS could be used as a beaconless wavefront sensor in a multiconjugate adaptive optics system when imaging extended scenes.

  14. Diffraction-limited IR Microspectroscopy with IRENI

    Treesearch

    J. Sedlmair; B. Illman; M. Unger; C. Hirschmugl

    2012-01-01

    In a unique way, IRENI (Infrared environmental Imaging), operated at the Synchrotron Radiation Center in Madison, combines IR spectroscopy and IR imaging, revealing the chemical morphology of a sample. Most storage ring based IR confocal microscopes have to overcome a trade-off between spatial resolution versus...

  15. Diffraction limited gamma-ray optics using Fresnel lenses for micro-arc second angular resolution

    NASA Astrophysics Data System (ADS)

    Skinner, G.; von Ballmoos, P.; Gehrels, N.; Krzmanic, J.

    2003-03-01

    Refractive indices at gamma-ray wavelengths are such that material thicknesses of the order of millimeters allow the phase of a wavefront to be changed by up to 2π . Thus a phase Fresnel lens can be made from a simple profiled thin disk of, for example, aluminium or plastic. Such a lens can easily have a collecting area of several square meters and an efficiency >90%. Ordinary engineering tolerances allow the manufacture of a lens which can be diffraction limited in the pico-meter wavelength band (up to ˜MeV) and thus provides a simple optical system with angular resolution better than a micro arc second i.e. the resolution necessary to resolve structures on the scale of the event horizon of super-massive black holes in AGN. However the focal length of such a lens is very long - up to a million km. Nevertheless studies have shown that a mission `Fresnel' using a detector and a phase Fresnel lens on two station-keeping spacecraft separated by such a distance is feasible. Results from these studies and work on other proof of concept studies are presented.

  16. Design and fabrication of a miniature objective consisting of high refractive index zinc sulfide lenses for laser surgery

    PubMed Central

    Shadfan, Adam; Pawlowski, Michal; Wang, Ye; Subramanian, Kaushik; Gabay, Ilan; Ben-Yakar, Adela; Tkaczyk, Tomasz

    2016-01-01

    A miniature laser ablation probe relying on an optical fiber to deliver light requires a high coupling efficiency objective with sufficient magnification in order to provide adequate power and field for surgery. A diffraction-limited optical design is presented that utilizes high refractive index zinc sulfide to meet specifications while reducing the miniature objective down to two lenses. The design has a hypercentric conjugate plane on the fiber side and is telecentric on the tissue end. Two versions of the objective were built on a diamond lathe—a traditional cylindrical design and a custom-tapered mount. Both received an antireflective coating. The objectives performed as designed in terms of observable resolution and field of view as measured by imaging a 1951 USAF resolution target. The slanted edge technique was used to find Strehl ratios of 0.75 and 0.78, respectively, indicating nearly diffraction-limited performance. Finally, preliminary ablation experiments indicated threshold fluence of gold film was comparable to similar reported probes. PMID:28579656

  17. Design and fabrication of a miniature objective consisting of high refractive index zinc sulfide lenses for laser surgery

    NASA Astrophysics Data System (ADS)

    Shadfan, Adam; Pawlowski, Michal; Wang, Ye; Subramanian, Kaushik; Gabay, Ilan; Ben-Yakar, Adela; Tkaczyk, Tomasz

    2016-02-01

    A miniature laser ablation probe relying on an optical fiber to deliver light requires a high coupling efficiency objective with sufficient magnification in order to provide adequate power and field for surgery. A diffraction-limited optical design is presented that utilizes high refractive index zinc sulfide to meet specifications while reducing the miniature objective down to two lenses. The design has a hypercentric conjugate plane on the fiber side and is telecentric on the tissue end. Two versions of the objective were built on a diamond lathe-a traditional cylindrical design and a custom-tapered mount. Both received an antireflective coating. The objectives performed as designed in terms of observable resolution and field of view as measured by imaging a 1951 USAF resolution target. The slanted edge technique was used to find Strehl ratios of 0.75 and 0.78, respectively, indicating nearly diffraction-limited performance. Finally, preliminary ablation experiments indicated threshold fluence of gold film was comparable to similar reported probes.

  18. Evolutionary optimization of compact dielectric lens for farfield sub-wavelength imaging

    PubMed Central

    Zhang, Jingjing

    2015-01-01

    The resolution of conventional optical lenses is limited by diffraction. For decades researchers have made various attempts to beat the diffraction limit and realize subwavelength imaging. Here we present the approach to design modified solid immersion lenses that deliver the subwavelength information of objects into the far field, yielding magnified images. The lens is composed of an isotropic dielectric core and anisotropic or isotropic dielectric matching layers. It is designed by combining a transformation optics forward design with an inverse design scheme, where an evolutionary optimization procedure is applied to find the material parameters for the matching layers. Notably, the total radius of the lens is only 2.5 wavelengths and the resolution can reach λ/6. Compared to previous approaches based on the simple discretized approximation of a coordinate transformation design, our method allows for much more precise recovery of the information of objects, especially for those with asymmetric shapes. It allows for the far-field subwavelength imaging at optical frequencies with compact dielectric devices. PMID:26017657

  19. Single-spin stochastic optical reconstruction microscopy

    PubMed Central

    Pfender, Matthias; Aslam, Nabeel; Waldherr, Gerald; Neumann, Philipp; Wrachtrup, Jörg

    2014-01-01

    We experimentally demonstrate precision addressing of single-quantum emitters by combined optical microscopy and spin resonance techniques. To this end, we use nitrogen vacancy (NV) color centers in diamond confined within a few ten nanometers as individually resolvable quantum systems. By developing a stochastic optical reconstruction microscopy (STORM) technique for NV centers, we are able to simultaneously perform sub–diffraction-limit imaging and optically detected spin resonance (ODMR) measurements on NV spins. This allows the assignment of spin resonance spectra to individual NV center locations with nanometer-scale resolution and thus further improves spatial discrimination. For example, we resolved formerly indistinguishable emitters by their spectra. Furthermore, ODMR spectra contain metrology information allowing for sub–diffraction-limit sensing of, for instance, magnetic or electric fields with inherently parallel data acquisition. As an example, we have detected nuclear spins with nanometer-scale precision. Finally, we give prospects of how this technique can evolve into a fully parallel quantum sensor for nanometer resolution imaging of delocalized quantum correlations. PMID:25267655

  20. Structured illumination for wide-field Raman imaging of cell membranes

    NASA Astrophysics Data System (ADS)

    Chen, Houkai; Wang, Siqi; Zhang, Yuquan; Yang, Yong; Fang, Hui; Zhu, Siwei; Yuan, Xiaocong

    2017-11-01

    Although the diffraction limit still restricts their lateral resolution, conventional wide-field Raman imaging techniques offer fast imaging speeds compared with scanning schemes. To extend the lateral resolution of wide-field Raman microscopy using filters, standing-wave illumination technique is used, and an improvement of lateral resolution by a factor of more than two is achieved. Specifically, functionalized surface enhanced Raman scattering nanoparticles are employed to strengthen the desired scattering signals to label cell membranes. This wide-field Raman imaging technique affords various significant opportunities in the biological applications.

  1. Understanding Super-Resolution Nanoscopy and Its Biological Applications in Cell Imaging

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hu, Dehong; Zhao, Baoming; Xie, Yumei

    2013-01-01

    Optical microscopy has been an ideal tool to study phenomena in live cells because visible light at reasonable intensity does not perturb much of the normal biological functions. However, optical resolution using visible light is significantly limited by the wavelength. Overcoming this diffraction-limit barrier will reveal biological mechanisms, cellular structures, and physiological processes at nanometer scale, orders of magnitude lower than current optical microscopy. Although this appears to be a daunting task, recently developed photoswitchable probes enable reconstruction of individual images into a super-resolution image, thus the emergence of nanoscopy. Harnessing the resolution power of nanoscopy, we report here nano-resolutionmore » fluorescence imaging of microtubules and their network structures in biological cells. The super-resolution nanoscopy successfully resolved nanostructures of microtubule network—a daunting task that cannot be completed using conventional wide-field microscopy.« less

  2. Optical super resolution using tilted illumination coupled with object rotation

    NASA Astrophysics Data System (ADS)

    Hussain, Anwar; Mudassar, Asloob A.

    2015-03-01

    In conventional imaging systems, the resolution of the final image is mainly distorted due to diffraction of higher spatial frequencies of the target object. To overcome the diffraction limit, imaging techniques which synthetically enlarge the aperture of the system are used. In this paper, synthesized aperture is produced by means of a three fiber illumination assembly coupled with an in-plane object rotation. The high order diffracted spatial frequencies of the object are brought into the pass band of optical system by illuminating the object with tilted beams. The tilt produced at the fiber assembly plane is related to the dimension of the aperture, placed at the Fourier plane of the system. To span the 2D object spectrum at the Fourier plane, an in-plane object rotation procedure is applied at the object plane. The spectrum of the object is rotated as the object is rotated and illuminated with tilted beams. The corresponding object beam is interfered with a reference beam from the same source to record interferograms. All the recorded interferograms are stored in computer and de-convolution algorithm is applied to recover the synthesized spectrum. The image of the synthesized spectrum has three times improved resolution compared to the conventional image.

  3. Analysis of the diffraction effects for a multi-view autostereoscopic three-dimensional display system based on shutter parallax barriers with full resolution

    NASA Astrophysics Data System (ADS)

    Meng, Yang; Yu, Zhongyuan; Jia, Fangda; Zhang, Chunyu; Wang, Ye; Liu, Yumin; Ye, Han; Chen, Laurence Lujun

    2017-10-01

    A multi-view autostereoscopic three-dimensional (3D) system is built by using a 2D display screen and a customized parallax-barrier shutter (PBS) screen. The shutter screen is controlled dynamically by address driving matrix circuit and it is placed in front of the display screen at a certain location. The system could achieve densest viewpoints due to its specially optical and geometric design which is based on concept of "eye space". The resolution of 3D imaging is not reduced compared to 2D mode by using limited time division multiplexing technology. The diffraction effects may play an important role in 3D display imaging quality, especially when applied to small screen, such as iPhone screen etc. For small screen, diffraction effects may contribute crosstalk between binocular views, image brightness uniformity etc. Therefore, diffraction effects are analyzed and considered in a one-dimensional shutter screen model of the 3D display, in which the numerical simulation of light from display pixels on display screen through parallax barrier slits to each viewing zone in eye space, is performed. The simulation results provide guidance for criteria screen size over which the impact of diffraction effects are ignorable, and below which diffraction effects must be taken into account. Finally, the simulation results are compared to the corresponding experimental measurements and observation with discussion.

  4. Adaptive optics high-resolution IR spectroscopy with silicon grisms and immersion gratings

    NASA Astrophysics Data System (ADS)

    Ge, Jian; McDavitt, Daniel L.; Chakraborty, Abhijit; Bernecker, John L.; Miller, Shane

    2003-02-01

    The breakthrough of silicon immersion grating technology at Penn State has the ability to revolutionize high-resolution infrared spectroscopy when it is coupled with adaptive optics at large ground-based telescopes. Fabrication of high quality silicon grism and immersion gratings up to 2 inches in dimension, less than 1% integrated scattered light, and diffraction-limited performance becomes a routine process thanks to newly developed techniques. Silicon immersion gratings with etched dimensions of ~ 4 inches are being developed at Penn State. These immersion gratings will be able to provide a diffraction-limited spectral resolution of R = 300,000 at 2.2 micron, or 130,000 at 4.6 micron. Prototype silicon grisms have been successfully used in initial scientific observations at the Lick 3m telescope with adaptive optics. Complete K band spectra of a total of 6 T Tauri and Ae/Be stars and their close companions at a spectral resolution of R ~ 3000 were obtained. This resolving power was achieved by using a silicon echelle grism with a 5 mm pupil diameter in an IR camera. These results represent the first scientific observations conducted by the high-resolution silicon grisms, and demonstrate the extremely high dispersing power of silicon-based gratings. New discoveries from this high spatial and spectral resolution IR spectroscopy will be reported. The future of silicon-based grating applications in ground-based AO IR instruments is promising. Silicon immersion gratings will make very high-resolution spectroscopy (R > 100,000) feasible with compact instruments for implementation on large telescopes. Silicon grisms will offer an efficient way to implement low-cost medium to high resolution IR spectroscopy (R ~ 1000-50000) through the conversion of existing cameras into spectrometers by locating a grism in the instrument's pupil location.

  5. Scanning near-field optical microscopy.

    PubMed

    Vobornik, Dusan; Vobornik, Slavenka

    2008-02-01

    An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today's science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM) is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution.

  6. Structured illumination microscopy for dual-modality 3D sub-diffraction resolution fluorescence and refractive-index reconstruction

    PubMed Central

    Chowdhury, Shwetadwip; Eldridge, Will J.; Wax, Adam; Izatt, Joseph A.

    2017-01-01

    Though structured illumination (SI) microscopy is a popular imaging technique conventionally associated with fluorescent super-resolution, recent works have suggested its applicability towards sub-diffraction resolution coherent imaging with quantitative endogenous biological contrast. Here, we demonstrate that SI can efficiently integrate together the principles of fluorescent super-resolution and coherent synthetic aperture to achieve 3D dual-modality sub-diffraction resolution, fluorescence and refractive-index (RI) visualizations of biological samples. We experimentally demonstrate this framework by introducing a SI microscope capable of 3D sub-diffraction resolution fluorescence and RI imaging, and verify its biological visualization capabilities by experimentally reconstructing 3D RI/fluorescence visualizations of fluorescent calibration microspheres as well as alveolar basal epithelial adenocarcinoma (A549) and human colorectal adenocarcinmoa (HT-29) cells, fluorescently stained for F-actin. This demonstration may suggest SI as an especially promising imaging technique to enable future biological studies that explore synergistically operating biophysical/biochemical and molecular mechanisms at sub-diffraction resolutions. PMID:29296504

  7. In-line digital holography with phase-shifting Greek-ladder sieves

    NASA Astrophysics Data System (ADS)

    Xie, Jing; Zhang, Junyong; Zhang, Yanli; Zhou, Shenlei; Zhu, Jianqiang

    2018-04-01

    Phase shifting is the key technique in in-line digital holography, but traditional phase shifters have their own limitations in short wavelength regions. Here, phase-shifting Greek-ladder sieves with amplitude-only modulation are introduced into in-line digital holography, which are essentially a kind of diffraction lens with three-dimensional array diffraction-limited foci. In the in-line digital holographic experiment, we design two kinds of sieves by lithography and verify the validity of their phase-shifting function by measuring a 1951 U.S. Air Force resolution test target and three-dimensional array foci. With advantages of high resolving power, low cost, and no limitations at shorter wavelengths, phase-shifting Greek-ladder sieves have great potential in X-ray holography or biochemical microscopy for the next generation of synchrotron light sources.

  8. Unusual scaling laws for plasmonic nanolasers beyond the diffraction limit.

    PubMed

    Wang, Suo; Wang, Xing-Yuan; Li, Bo; Chen, Hua-Zhou; Wang, Yi-Lun; Dai, Lun; Oulton, Rupert F; Ma, Ren-Min

    2017-12-01

    Plasmonic nanolasers are a new class of amplifiers that generate coherent light well below the diffraction barrier bringing fundamentally new capabilities to biochemical sensing, super-resolution imaging, and on-chip optical communication. However, a debate about whether metals can enhance the performance of lasers has persisted due to the unavoidable fact that metallic absorption intrinsically scales with field confinement. Here, we report plasmonic nanolasers with extremely low thresholds on the order of 10 kW cm -2 at room temperature, which are comparable to those found in modern laser diodes. More importantly, we find unusual scaling laws allowing plasmonic lasers to be more compact and faster with lower threshold and power consumption than photonic lasers when the cavity size approaches or surpasses the diffraction limit. This clarifies the long-standing debate over the viability of metal confinement and feedback strategies in laser technology and identifies situations where plasmonic lasers can have clear practical advantage.

  9. “MODAL NOISE” IN SINGLE-MODE FIBERS: A CAUTIONARY NOTE FOR HIGH PRECISION RADIAL VELOCITY INSTRUMENTS

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Halverson, Samuel; Roy, Arpita; Mahadevan, Suvrath

    2015-12-01

    Exploring the use of single-mode fibers (SMFs) in high precision Doppler spectrometers has become increasingly attractive since the advent of diffraction-limited adaptive optics systems on large-aperture telescopes. Spectrometers fed with these fibers can be made significantly smaller than typical “seeing-limited” instruments, greatly reducing cost and overall complexity. Importantly, classical mode interference and speckle issues associated with multi-mode fibers, also known as “modal noise,” are mitigated when using SMFs, which also provide perfect radial and azimuthal image scrambling. However, SMFs do support multiple polarization modes, an issue that is generally ignored for larger-core fibers given the large number of propagation modes.more » Since diffraction gratings used in most high resolution astronomical instruments have dispersive properties that are sensitive to incident polarization changes, any birefringence variations in the fiber can cause variations in the efficiency profile, degrading illumination stability. Here we present a cautionary note outlining how the polarization properties of SMFs can affect the radial velocity (RV) measurement precision of high resolution spectrographs. This work is immediately relevant to the rapidly expanding field of diffraction-limited, extreme precision RV spectrographs that are currently being designed and built by a number of groups.« less

  10. Mapping atomic motions with ultrabright electrons: towards fundamental limits in space-time resolution.

    PubMed

    Manz, Stephanie; Casandruc, Albert; Zhang, Dongfang; Zhong, Yinpeng; Loch, Rolf A; Marx, Alexander; Hasegawa, Taisuke; Liu, Lai Chung; Bayesteh, Shima; Delsim-Hashemi, Hossein; Hoffmann, Matthias; Felber, Matthias; Hachmann, Max; Mayet, Frank; Hirscht, Julian; Keskin, Sercan; Hada, Masaki; Epp, Sascha W; Flöttmann, Klaus; Miller, R J Dwayne

    2015-01-01

    The long held objective of directly observing atomic motions during the defining moments of chemistry has been achieved based on ultrabright electron sources that have given rise to a new field of atomically resolved structural dynamics. This class of experiments requires not only simultaneous sub-atomic spatial resolution with temporal resolution on the 100 femtosecond time scale but also has brightness requirements approaching single shot atomic resolution conditions. The brightness condition is in recognition that chemistry leads generally to irreversible changes in structure during the experimental conditions and that the nanoscale thin samples needed for electron structural probes pose upper limits to the available sample or "film" for atomic movies. Even in the case of reversible systems, the degree of excitation and thermal effects require the brightest sources possible for a given space-time resolution to observe the structural changes above background. Further progress in the field, particularly to the study of biological systems and solution reaction chemistry, requires increased brightness and spatial coherence, as well as an ability to tune the electron scattering cross-section to meet sample constraints. The electron bunch density or intensity depends directly on the magnitude of the extraction field for photoemitted electron sources and electron energy distribution in the transverse and longitudinal planes of electron propagation. This work examines the fundamental limits to optimizing these parameters based on relativistic electron sources using re-bunching cavity concepts that are now capable of achieving 10 femtosecond time scale resolution to capture the fastest nuclear motions. This analysis is given for both diffraction and real space imaging of structural dynamics in which there are several orders of magnitude higher space-time resolution with diffraction methods. The first experimental results from the Relativistic Electron Gun for Atomic Exploration (REGAE) are given that show the significantly reduced multiple electron scattering problem in this regime, which opens up micron scale systems, notably solution phase chemistry, to atomically resolved structural dynamics.

  11. Review of ultra-high density optical storage technologies for big data center

    NASA Astrophysics Data System (ADS)

    Hao, Ruan; Liu, Jie

    2016-10-01

    In big data center, optical storage technologies have many advantages, such as energy saving and long lifetime. However, how to improve the storage density of optical storage is still a huge challenge. Maybe the multilayer optical storage technology is the good candidate for big data center in the years to come. Due to the number of layers is primarily limited by transmission of each layer, the largest capacities of the multilayer disc are around 1 TB/disc and 10 TB/ cartridge. Holographic data storage (HDS) is a volumetric approach, but its storage capacity is also strictly limited by the diffractive nature of light. For a holographic disc with total thickness of 1.5mm, its potential capacities are not more than 4TB/disc and 40TB/ cartridge. In recent years, the development of super resolution optical storage technology has attracted more attentions. Super-resolution photoinduction-inhibition nanolithography (SPIN) technology with 9 nm feature size and 52nm two-line resolution was reported 3 years ago. However, turning this exciting principle into a real storage system is a huge challenge. It can be expected that in the future, the capacities of 10TB/disc and 100TB/cartridge can be achieved. More importantly, due to breaking the diffraction limit of light, SPIN technology will open the door to improve the optical storage capacity steadily to meet the need of the developing big data center.

  12. Sub-Millisecond Time Resolved X-ray Surface Diffraction During Pulsed Laser Deposition

    NASA Astrophysics Data System (ADS)

    Tischler, J. Z.; Larson, B. C.; Eres, Gyula; Rouleau, C. M.; Lowndes, D. H.; Yoon, M.; Zschack, P.

    2001-03-01

    The initial crystallization and evolution of the SrTiO3 (001) surface during homoeptaxial pulsed laser deposition growth of SrTiO3 was studied using time resolved surface x-ray diffraction with a time resolution down to 200 μ s. Measurements performed at the UNICAT undulator line at the Advanced Photon Source indicated prompt formation of epitaxial SrTiO3 bi-layers down to our limiting time resolution. The subsequent evolution of the surface occurred on a much greater time scale, and was studied both by measurements of surface truncation rod intensities and by measurements of diffuse scattering near the rod. The effect of temperature and correlation with in-plane order will also be discussed.

  13. Diffractive X-Ray Telescopes

    NASA Technical Reports Server (NTRS)

    Skinner, Gerald K.

    2010-01-01

    Diffractive X-ray telescopes, using zone plates, phase Fresnel lenses, or related optical elements have the potential to provide astronomers with true imaging capability with resolution many orders of magnitude better than available in any other waveband. Lenses that would be relatively easy to fabricate could have an angular resolution of the order of micro-arc-seconds or even better, that would allow, for example, imaging of the distorted spacetime in the immediate vicinity of the super-massive black holes in the center of active galaxies. What then is precluding their immediate adoption? Extremely long focal lengths, very limited bandwidth, and difficulty stabilizing the image are the main problems. The history, and status of the development of such lenses is reviewed here and the prospects for managing the challenges that they present are discussed.

  14. Three-dimensional electron diffraction of plant light-harvesting complex

    PubMed Central

    Wang, Da Neng; Kühlbrandt, Werner

    1992-01-01

    Electron diffraction patterns of two-dimensional crystals of light-harvesting chlorophyll a/b-protein complex (LHC-II) from photosynthetic membranes of pea chloroplasts, tilted at different angles up to 60°, were collected to 3.2 Å resolution at -125°C. The reflection intensities were merged into a three-dimensional data set. The Friedel R-factor and the merging R-factor were 21.8 and 27.6%, respectively. Specimen flatness and crystal size were critical for recording electron diffraction patterns from crystals at high tilts. The principal sources of experimental error were attributed to limitations of the number of unit cells contributing to an electron diffraction pattern, and to the critical electron dose. The distribution of strong diffraction spots indicated that the three-dimensional structure of LHC-II is less regular than that of other known membrane proteins and is not dominated by a particular feature of secondary structure. ImagesFIGURE 1FIGURE 2 PMID:19431817

  15. High quality transmission Kikuchi diffraction analysis of deformed alloys - Case study

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tokarski, Tomasz, E-mail: tokarski@agh.edu.pl

    Modern scanning electron microscopes (SEM) equipped with thermally assisted field emission guns (Schottky FEG) are capable of imaging with a resolution in the range of several nanometers or better. Simultaneously, the high electron beam current can be used, which enables fast chemical and crystallographic analysis with a higher resolution than is normally offered by SEM with a tungsten cathode. The current resolution that limits the EDS and EBSD analysis is related to materials' physics, particularly to the electron-specimen interaction volume. The application of thin, electron-transparent specimens, instead of bulk samples, improves the resolution and allows for the detailed analysis ofmore » very fine microstructural features. Beside the typical imaging mode, it is possible to use a standard EBSD camera in such a configuration that only transmitted and scattered electrons are detected. This modern approach was successfully applied to various materials giving rise to significant resolution improvement, especially for the light element magnesium based alloys. This paper presents an insight into the application of the transmission Kikuchi diffraction (TKD) technique applied to the most troublesome, heavily-deformed materials. In particular, the values of the highest possible acquisition rates for high resolution and high quality mapping were estimated within typical imaging conditions of stainless steel and magnesium-yttrium alloy. - Highlights: •Monte Carlo simulations were used to simulate EBSD camera intensity for various measuring conditions. •Transmission Kikuchi diffraction parameters were evaluated for highly deformed, light and heavy elements based alloys. •High quality maps with 20 nm spatial resolution were acquired for Mg and Fe based alloys. •High speed TKD measurements were performed at acquisition rates comparable to the reflection EBSD.« less

  16. Correlative Single-Molecule Localization Microscopy and Confocal Microscopy.

    PubMed

    Soeller, Christian; Hou, Yufeng; Jayasinghe, Isuru D; Baddeley, David; Crossman, David

    2017-01-01

    Single-molecule localization microscopy allows the ability to image fluorescence labeled molecular targets at nanoscale resolution. However, for many biological questions the ability to provide tissue and cellular context in addition to these high resolution data is eminently informative. Here, we describe a procedure to achieve this aim by correlatively imaging human cardiac tissue first at the nanoscale with direct stochastic optical reconstruction microscopy (dSTORM) and then at the diffraction limit with conventional confocal microscopy.

  17. Optical system analysis for the ground based EXVM

    NASA Technical Reports Server (NTRS)

    Hillman, L. W.; Chipman, R. A.; Smith, M. H.

    1993-01-01

    The MSFC's Experimental Vector Magnetograph (EXVM) is an instrument that observes a 4.4 x 8.8 arcmin field of the sun. The transverse and longitudinal components of the surface magnetic field and the line-of-sight velocities of the photospheric gases can be determined from polarimetric and spectral analysis of the 525.02 nm absorption line of Fe 1. The EXVM has been breadboarded and tested in the laboratory. The optics of the EXVM were tested with a point-diffraction (Smartt) interferometer. The 12 inch Cassegrain telescope was found to have 0.20 waves RMS (at 525.02 nm) of aberration. The post-telescope relay optics were nearly diffraction limited on-axis and had about one wave of primary coma as the predominant aberration at full-field. From theoretical modulation transfer function (MTF) curves of known aberrations, it was concluded that the EXVM should attain a maximum spatial resolution of about 0.5 arcseconds. A resolution test target indicated maximum angular resolutions better than 0.6 arcsec on-axis and 0.7 arcsec at full-field-of-view. A 2D inch heliostat (sun-tracking mirror) was used to direct sunlight into the lab and into the EXVM. Solar images obtained were limited by atmospheric seeing effects. During brief moments of good seeing, angular resolutions of about 1 arcsecond were realized with the EXVM.

  18. Plasmonics and metamaterials based super-resolution imaging (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Liu, Zhaowei

    2017-05-01

    In recent years, surface imaging of various biological dynamics and biomechanical phenomena has seen a surge of interest. Imaging of processes such as exocytosis and kinesin motion are most effective when depth is limited to a very thin region of interest at the edge of the cell or specimen. However, many objects and processes of interest are of size scales below the diffraction limit for safe, visible wavelength illumination. Super-resolution imaging methods such as structured illumination microscopy and others have offered various compromises between resolution, imaging speed, and bio-compatibility. In this talk, I will present our most recent progress in plasmonic structured illumination microscopy (PSIM) and localized plasmonic structured illumination microscopy (LPSIM), and their applications in bio-imaging. We have achieved wide-field surface imaging with resolution down to 75 nm while maintaining reasonable speed and compatibility with biological specimens. These plasmonic enhanced super resolution techniques offer unique solutions to obtain 50nm spatial resolution and 50 frames per second wide imaging speed at the same time.

  19. Optical design considerations when imaging the fundus with an adaptive optics correction

    NASA Astrophysics Data System (ADS)

    Wang, Weiwei; Campbell, Melanie C. W.; Kisilak, Marsha L.; Boyd, Shelley R.

    2008-06-01

    Adaptive Optics (AO) technology has been used in confocal scanning laser ophthalmoscopes (CSLO) which are analogous to confocal scanning laser microscopes (CSLM) with advantages of real-time imaging, increased image contrast, a resistance to image degradation by scattered light, and improved optical sectioning. With AO, the instrumenteye system can have low enough aberrations for the optical quality to be limited primarily by diffraction. Diffraction-limited, high resolution imaging would be beneficial in the understanding and early detection of eye diseases such as diabetic retinopathy. However, to maintain diffraction-limited imaging, sufficient pixel sampling over the field of view is required, resulting in the need for increased data acquisition rates for larger fields. Imaging over smaller fields may be a disadvantage with clinical subjects because of fixation instability and the need to examine larger areas of the retina. Reduction in field size also reduces the amount of light sampled per pixel, increasing photon noise. For these reasons, we considered an instrument design with a larger field of view. When choosing scanners to be used in an AOCSLO, the ideal frame rate should be above the flicker fusion rate for the human observer and would also allow user control of targets projected onto the retina. In our AOCSLO design, we have studied the tradeoffs between field size, frame rate and factors affecting resolution. We will outline optical approaches to overcome some of these tradeoffs and still allow detection of the earliest changes in the fundus in diabetic retinopathy.

  20. Continuous motion scan ptychography: Characterization for increased speed in coherent x-ray imaging

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Deng, Junjing; Nashed, Youssef S. G.; Chen, Si

    Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object’s complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous “fly-scan” mode for ptychographic data collection in whichmore » the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.« less

  1. Continuous motion scan ptychography: Characterization for increased speed in coherent x-ray imaging

    DOE PAGES

    Deng, Junjing; Nashed, Youssef S. G.; Chen, Si; ...

    2015-02-23

    Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object’s complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous “fly-scan” mode for ptychographic data collection in whichmore » the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.« less

  2. Continuous motion scan ptychography: characterization for increased speed in coherent x-ray imaging

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Deng, Junjing; Nashed, Youssef S. G.; Chen, Si

    2015-01-01

    Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object's complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous "fly-scan" mode for ptychographic data collection in whichmore » the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.« less

  3. Continuous motion scan ptychography: characterization for increased speed in coherent x-ray imaging.

    PubMed

    Deng, Junjing; Nashed, Youssef S G; Chen, Si; Phillips, Nicholas W; Peterka, Tom; Ross, Rob; Vogt, Stefan; Jacobsen, Chris; Vine, David J

    2015-03-09

    Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object's complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous "fly-scan" mode for ptychographic data collection in which the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.

  4. Nanoscale volume confinement and fluorescence enhancement with double nanohole aperture

    PubMed Central

    Regmi, Raju; Al Balushi, Ahmed A.; Rigneault, Hervé; Gordon, Reuven; Wenger, Jérôme

    2015-01-01

    Diffraction ultimately limits the fluorescence collected from a single molecule, and sets an upper limit to the maximum concentration to isolate a single molecule in the detection volume. To overcome these limitations, we introduce here the use of a double nanohole structure with 25 nm gap, and report enhanced detection of single fluorescent molecules in concentrated solutions exceeding 20 micromolar. The nanometer gap concentrates the light into an apex volume down to 70 zeptoliter (10−21 L), 7000-fold below the diffraction-limited confocal volume. Using fluorescence correlation spectroscopy and time-correlated photon counting, we measure fluorescence enhancement up to 100-fold, together with local density of optical states (LDOS) enhancement of 30-fold. The distinctive features of double nanoholes combining high local field enhancement, efficient background screening and relative nanofabrication simplicity offer new strategies for real time investigation of biochemical events with single molecule resolution at high concentrations. PMID:26511149

  5. Liquid Crystal on Silicon Wavefront Corrector

    NASA Technical Reports Server (NTRS)

    Pouch, John; Miranda, Felix; Wang, Xinghua; Bos, Philip, J.

    2004-01-01

    A low cost, high resolution, liquid crystal on silicon, spatial light modulator has been developed for the correction of huge aberrations in an optical system where the polarization dependence and the chromatic nature are tolerated. However, the overall system performance suggests that this device is also suitable for real time correction of aberration in human eyes. This device has a resolution of 1024 x 768, and is driven by an XGA display driver. The effective stroke length of the device is 700 nm and 2000 nm for the visible and IR regions of the device, respectively. The response speeds are 50 Hz and 5 Hz, respectively, which are fast enough for real time adaptive optics for aberrations in human eyes. By modulating a wavefront of 2 pi, this device can correct for arbitrary high order wavefront aberrations since the 2-D pixel array is independently controlled by the driver. The high resolution and high accuracy of the device allow for diffraction limited correction of the tip and tilt or defocus without an additional correction loop. We have shown that for every wave of aberration, an 8 step blazed grating is required to achieve high diffraction efficiency around 80%. In light of this, up to 125 waves peak to valley of tip and tilt can be corrected if we choose the simplest aberration. Corrections of 34 waves of aberration, including high order Zernicke terms in a high magnification telescope, to diffraction limited performance (residual wavefront aberration less than 1/30 lambda at 632.8 nm) have been observed at high efficiency.

  6. Wide field of view 3D label-free super-resolution imaging

    NASA Astrophysics Data System (ADS)

    Nolvi, Anton; Laidmäe, Ivo; Maconi, Göran; Heinämäki, Jyrki; Hæggström, Edward; Kassamakov, Ivan

    2018-02-01

    Recently, 3D label-free super-resolution profilers based on microsphere-assisted scanning white light interferometry were introduced having vertical resolution of few angstroms (Å) and a lateral resolution approaching 100 nm. However, the use of a single microsphere to generate the photonic nanojet (PNJ) limits their field of view. We overcome this limitation by using polymer microfibers to generate the PNJ. This increases the field of view by order of magnitude in comparison to the previously developed solutions while still resolving sub 100 nm features laterally and keeping the vertical resolution in 1nm range. To validate the capabilities of our system we used a recordable Blu-ray disc as a sample. It features a grooved surface topology with heights in the range of 20 nm and with distinguishable sub 100 nm lateral features that are unresolvable by diffraction limited optics. We achieved agreement between all three measurement devices across lateral and vertical dimensions. The field of view of our instrument was 110 μm by 2 μm and the imaging time was a couple of seconds.

  7. Chemically Resolved Imaging of Biological Cells and Thin Films by Infrared Scanning Near-Field Optical Microscopy

    PubMed Central

    Cricenti, Antonio; Generosi, Renato; Luce, Marco; Perfetti, Paolo; Margaritondo, Giorgio; Talley, David; Sanghera, Jas S.; Aggarwal, Ishwar D.; Tolk, Norman H.; Congiu-Castellano, Agostina; Rizzo, Mark A.; Piston, David W.

    2003-01-01

    The infrared (IR) absorption of a biological system can potentially report on fundamentally important microchemical properties. For example, molecular IR profiles are known to change during increases in metabolic flux, protein phosphorylation, or proteolytic cleavage. However, practical implementation of intracellular IR imaging has been problematic because the diffraction limit of conventional infrared microscopy results in low spatial resolution. We have overcome this limitation by using an IR spectroscopic version of scanning near-field optical microscopy (SNOM), in conjunction with a tunable free-electron laser source. The results presented here clearly reveal different chemical constituents in thin films and biological cells. The space distribution of specific chemical species was obtained by taking SNOM images at IR wavelengths (λ) corresponding to stretch absorption bands of common biochemical bonds, such as the amide bond. In our SNOM implementation, this chemical sensitivity is combined with a lateral resolution of 0.1 μm (≈λ/70), well below the diffraction limit of standard infrared microscopy. The potential applications of this approach touch virtually every aspect of the life sciences and medical research, as well as problems in materials science, chemistry, physics, and environmental research. PMID:14507733

  8. Imaging cervical cytology with scanning near-field optical microscopy (SNOM) coupled with an IR-FEL.

    PubMed

    Halliwell, Diane E; Morais, Camilo L M; Lima, Kássio M G; Trevisan, Julio; Siggel-King, Michele R F; Craig, Tim; Ingham, James; Martin, David S; Heys, Kelly A; Kyrgiou, Maria; Mitra, Anita; Paraskevaidis, Evangelos; Theophilou, Georgios; Martin-Hirsch, Pierre L; Cricenti, Antonio; Luce, Marco; Weightman, Peter; Martin, Francis L

    2016-07-12

    Cervical cancer remains a major cause of morbidity and mortality among women, especially in the developing world. Increased synthesis of proteins, lipids and nucleic acids is a pre-condition for the rapid proliferation of cancer cells. We show that scanning near-field optical microscopy, in combination with an infrared free electron laser (SNOM-IR-FEL), is able to distinguish between normal and squamous low-grade and high-grade dyskaryosis, and between normal and mixed squamous/glandular pre-invasive and adenocarcinoma cervical lesions, at designated wavelengths associated with DNA, Amide I/II and lipids. These findings evidence the promise of the SNOM-IR-FEL technique in obtaining chemical information relevant to the detection of cervical cell abnormalities and cancer diagnosis at spatial resolutions below the diffraction limit (≥0.2 μm). We compare these results with analyses following attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy; although this latter approach has been demonstrated to detect underlying cervical atypia missed by conventional cytology, it is limited by a spatial resolution of ~3 μm to 30 μm due to the optical diffraction limit.

  9. Imaging cervical cytology with scanning near-field optical microscopy (SNOM) coupled with an IR-FEL

    PubMed Central

    Halliwell, Diane E.; Morais, Camilo L. M.; Lima, Kássio M. G.; Trevisan, Julio; Siggel-King, Michele R. F.; Craig, Tim; Ingham, James; Martin, David S.; Heys, Kelly A.; Kyrgiou, Maria; Mitra, Anita; Paraskevaidis, Evangelos; Theophilou, Georgios; Martin-Hirsch, Pierre L.; Cricenti, Antonio; Luce, Marco; Weightman, Peter; Martin, Francis L.

    2016-01-01

    Cervical cancer remains a major cause of morbidity and mortality among women, especially in the developing world. Increased synthesis of proteins, lipids and nucleic acids is a pre-condition for the rapid proliferation of cancer cells. We show that scanning near-field optical microscopy, in combination with an infrared free electron laser (SNOM-IR-FEL), is able to distinguish between normal and squamous low-grade and high-grade dyskaryosis, and between normal and mixed squamous/glandular pre-invasive and adenocarcinoma cervical lesions, at designated wavelengths associated with DNA, Amide I/II and lipids. These findings evidence the promise of the SNOM-IR-FEL technique in obtaining chemical information relevant to the detection of cervical cell abnormalities and cancer diagnosis at spatial resolutions below the diffraction limit (≥0.2 μm). We compare these results with analyses following attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy; although this latter approach has been demonstrated to detect underlying cervical atypia missed by conventional cytology, it is limited by a spatial resolution of ~3 μm to 30 μm due to the optical diffraction limit. PMID:27406404

  10. A Novel Modified Omega-K Algorithm for Synthetic Aperture Imaging Lidar through the Atmosphere

    PubMed Central

    Guo, Liang; Xing, Mendao; Tang, Yu; Dan, Jing

    2008-01-01

    The spatial resolution of a conventional imaging lidar system is constrained by the diffraction limit of the telescope's aperture. The combination of the lidar and synthetic aperture (SA) processing techniques may overcome the diffraction limit and pave the way for a higher resolution air borne or space borne remote sensor. Regarding the lidar transmitting frequency modulation continuous-wave (FMCW) signal, the motion during the transmission of a sweep and the reception of the corresponding echo were expected to be one of the major problems. The given modified Omega-K algorithm takes the continuous motion into account, which can compensate for the Doppler shift induced by the continuous motion efficiently and azimuth ambiguity for the low pulse recurrence frequency limited by the tunable laser. And then, simulation of Phase Screen (PS) distorted by atmospheric turbulence following the von Karman spectrum by using Fourier Transform is implemented in order to simulate turbulence. Finally, the computer simulation shows the validity of the modified algorithm and if in the turbulence the synthetic aperture length does not exceed the similar coherence length of the atmosphere for SAIL, we can ignore the effect of the turbulence. PMID:27879865

  11. Fast algorithm for chirp transforms with zooming-in ability and its applications.

    PubMed

    Deng, X; Bihari, B; Gan, J; Zhao, F; Chen, R T

    2000-04-01

    A general fast numerical algorithm for chirp transforms is developed by using two fast Fourier transforms and employing an analytical kernel. This new algorithm unifies the calculations of arbitrary real-order fractional Fourier transforms and Fresnel diffraction. Its computational complexity is better than a fast convolution method using Fourier transforms. Furthermore, one can freely choose the sampling resolutions in both x and u space and zoom in on any portion of the data of interest. Computational results are compared with analytical ones. The errors are essentially limited by the accuracy of the fast Fourier transforms and are higher than the order 10(-12) for most cases. As an example of its application to scalar diffraction, this algorithm can be used to calculate near-field patterns directly behind the aperture, 0 < or = z < d2/lambda. It compensates another algorithm for Fresnel diffraction that is limited to z > d2/lambdaN [J. Opt. Soc. Am. A 15, 2111 (1998)]. Experimental results from waveguide-output microcoupler diffraction are in good agreement with the calculations.

  12. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Küchemann, Stefan; Mahn, Carsten; Samwer, Konrad

    The investigation of short time dynamics using X-ray scattering techniques is commonly limited either by the read out frequency of the detector or by a low intensity. In this paper, we present a chopper system, which can increase the temporal resolution of 2D X-ray detectors by a factor of 13. This technique only applies to amorphous or polycrystalline samples due to their circular diffraction patterns. Using the chopper, we successfully increased the temporal resolution up to 5.1 ms during synchrotron experiments. For the construction, we provide a mathematical formalism, which, in principle, allows an even higher increase of the temporalmore » resolution.« less

  13. SPECKLE IMAGING EXCLUDES LOW-MASS COMPANIONS ORBITING THE EXOPLANET HOST STAR TRAPPIST-1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Howell, Steve B.; Scott, Nicholas J.; Everett, Mark E.

    2016-09-20

    We have obtained the highest-resolution images available of TRAPPIST-1 using the Gemini-South telescope and our speckle imaging camera. Observing at 692 and 883 nm, we reached the diffraction limit of the telescope providing a best resolution of 27 mas or, at the distance of TRAPPIST-1, a spatial resolution of 0.32 au. Our imaging of the star extends from 0.32 to 14.5 au. We show that to a high confidence level, we can exclude all possible stellar and brown dwarf companions, indicating that TRAPPIST-1 is a single star.

  14. Nature's crucible: Manufacturing optical nonlinearities for high resolution, high sensitivity encoding in the compound eye of the fly, Musca domestica

    NASA Technical Reports Server (NTRS)

    Wilcox, Mike

    1993-01-01

    The number of pixels per unit area sampling an image determines Nyquist resolution. Therefore, the highest pixel density is the goal. Unfortunately, as reduction in pixel size approaches the wavelength of light, sensitivity is lost and noise increases. Animals face the same problems and have achieved novel solutions. Emulating these solutions offers potentially unlimited sensitivity with detector size approaching the diffraction limit. Once an image is 'captured', cellular preprocessing of information allows extraction of high resolution information from the scene. Computer simulation of this system promises hyperacuity for machine vision.

  15. SCANNING NEAR-FIELD OPTICAL MICROSCOPY

    PubMed Central

    Vobornik, Dušan; Vobornik, Slavenka

    2008-01-01

    An average human eye can see details down to 0,07 mm in size. The ability to see smaller details of the matter is correlated with the development of the science and the comprehension of the nature. Today’s science needs eyes for the nano-world. Examples are easily found in biology and medical sciences. There is a great need to determine shape, size, chemical composition, molecular structure and dynamic properties of nano-structures. To do this, microscopes with high spatial, spectral and temporal resolution are required. Scanning Near-field Optical Microscopy (SNOM) is a new step in the evolution of microscopy. The conventional, lens-based microscopes have their resolution limited by diffraction. SNOM is not subject to this limitation and can offer up to 70 times better resolution. PMID:18318675

  16. Very high-resolution spectroscopy for extremely large telescopes using pupil slicing and adaptive optics.

    PubMed

    Beckers, Jacques M; Andersen, Torben E; Owner-Petersen, Mette

    2007-03-05

    Under seeing limited conditions very high resolution spectroscopy becomes very difficult for extremely large telescopes (ELTs). Using adaptive optics (AO) the stellar image size decreases proportional with the telescope diameter. This makes the spectrograph optics and hence its resolution independent of the telescope diameter. However AO for use with ELTs at visible wavelengths require deformable mirrors with many elements. Those are not likely to be available for quite some time. We propose to use the pupil slicing technique to create a number of sub-pupils each of which having its own deformable mirror. The images from all sub-pupils are combined incoherently with a diameter corresponding to the diffraction limit of the sub-pupil. The technique is referred to as "Pupil Slicing Adaptive Optics" or PSAO.

  17. Fabrication method of two-photon luminescent organic nano-architectures using electron-beam irradiation

    NASA Astrophysics Data System (ADS)

    Kamura, Yoshio; Imura, Kohei

    2018-06-01

    Optical recording on organic thin films with a high spatial resolution is promising for high-density optical memories, optical computing, and security systems. The spatial resolution of the optical recording is limited by the diffraction of light. Electrons can be focused to a nanometer-sized spot, providing the potential for achieving better resolution. In conventional electron-beam lithography, however, optical tuning of the fabricated structures is limited mostly to metals and semiconductors rather than organic materials. In this article, we report a fabrication method of luminescent organic architectures using a focused electron beam. We optimized the fabrication conditions of the electron beam to generate chemical species showing visible photoluminescence via two-photon near-infrared excitations. We utilized this fabrication method to draw nanoscale optical architectures on a polystyrene thin film.

  18. Ultrafast electron crystallography: Transient structures of molecules, surfaces, and phase transitions

    PubMed Central

    Ruan, Chong-Yu; Vigliotti, Franco; Lobastov, Vladimir A.; Chen, Songye; Zewail, Ahmed H.

    2004-01-01

    The static structure of macromolecular assemblies can be mapped out with atomic-scale resolution by using electron diffraction and microscopy of crystals. For transient nonequilibrium structures, which are critical to the understanding of dynamics and mechanisms, both spatial and temporal resolutions are required; the shortest scales of length (0.1–1 nm) and time (10–13 to 10–12 s) represent the quantum limit, the nonstatistical regime of rates. Here, we report the development of ultrafast electron crystallography for direct determination of structures with submonolayer sensitivity. In these experiments, we use crystalline silicon as a template for different adsorbates: hydrogen, chlorine, and trifluoroiodomethane. We observe the coherent restructuring of the surface layers with subangstrom displacement of atoms after the ultrafast heat impulse. This nonequilibrium dynamics, which is monitored in steps of 2 ps (total change ≤10 ps), contrasts that of the nanometer substrate. The effect of adsorbates and the phase transition at higher fluences were also studied through the evolution of streaks of interferences, Bragg spots (and their rocking curves), and rings in the diffraction patterns. We compare these results with kinematical theory and those of x-ray diffraction developed to study bulk behaviors. The sensitivity achieved here, with the 6 orders of magnitude larger cross section than x-ray diffraction, and with the capabilities of combined spatial (≈0.01 Å) and temporal (300–600 fs) resolutions, promise diverse applications for this ultrafast electron crystallography tabletop methodology. PMID:14745037

  19. OBSERVATIONS OF BINARY STARS WITH THE DIFFERENTIAL SPECKLE SURVEY INSTRUMENT. IV. OBSERVATIONS OF KEPLER, CoRoT, AND HIPPARCOS STARS FROM THE GEMINI NORTH TELESCOPE

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Horch, Elliott P.; Howell, Steve B.; Everett, Mark E.

    2012-12-01

    We present the results of 71 speckle observations of binary and unresolved stars, most of which were observed with the DSSI speckle camera at the Gemini North Telescope in 2012 July. The main purpose of the run was to obtain diffraction-limited images of high-priority targets for the Kepler and CoRoT missions, but in addition, we observed a number of close binary stars where the resolution limit of Gemini was used to better determine orbital parameters and/or confirm results obtained at or below the diffraction limit of smaller telescopes. Five new binaries and one triple system were discovered, and first orbitsmore » are calculated for other two systems. Several systems are discussed in detail.« less

  20. Diffraction study of duty-cycle error in ferroelectric quasi-phase-matching gratings with Gaussian beam illumination

    NASA Astrophysics Data System (ADS)

    Dwivedi, Prashant Povel; Kumar, Challa Sesha Sai Pavan; Choi, Hee Joo; Cha, Myoungsik

    2016-02-01

    Random duty-cycle error (RDE) is inherent in the fabrication of ferroelectric quasi-phase-matching (QPM) gratings. Although a small RDE may not affect the nonlinearity of QPM devices, it enhances non-phase-matched parasitic harmonic generations, limiting the device performance in some applications. Recently, we demonstrated a simple method for measuring the RDE in QPM gratings by analyzing the far-field diffraction pattern obtained by uniform illumination (Dwivedi et al. in Opt Express 21:30221-30226, 2013). In the present study, we used a Gaussian beam illumination for the diffraction experiment to measure noise spectra that are less affected by the pedestals of the strong diffraction orders. Our results were compared with our calculations based on a random grating model, demonstrating improved resolution in the RDE estimation.

  1. Effect of screw threading dislocations and inverse domain boundaries in GaN on the shape of reciprocal-space maps.

    PubMed

    Barchuk, Mykhailo; Motylenko, Mykhaylo; Lukin, Gleb; Pätzold, Olf; Rafaja, David

    2017-04-01

    The microstructure of polar GaN layers, grown by upgraded high-temperature vapour phase epitaxy on [001]-oriented sapphire substrates, was studied by means of high-resolution X-ray diffraction and transmission electron microscopy. Systematic differences between reciprocal-space maps measured by X-ray diffraction and those which were simulated for different densities of threading dislocations revealed that threading dislocations are not the only microstructure defect in these GaN layers. Conventional dark-field transmission electron microscopy and convergent-beam electron diffraction detected vertical inversion domains as an additional microstructure feature. On a series of polar GaN layers with different proportions of threading dislocations and inversion domain boundaries, this contribution illustrates the capability and limitations of coplanar reciprocal-space mapping by X-ray diffraction to distinguish between these microstructure features.

  2. High-resolution imaging of the Pluto-Charon system with the Faint Object Camera of the Hubble Space Telescope

    NASA Technical Reports Server (NTRS)

    Albrecht, R.; Barbieri, C.; Adorf, H.-M.; Corrain, G.; Gemmo, A.; Greenfield, P.; Hainaut, O.; Hook, R. N.; Tholen, D. J.; Blades, J. C.

    1994-01-01

    Images of the Pluto-Charon system were obtained with the Faint Object Camera (FOC) of the Hubble Space Telescope (HST) after the refurbishment of the telescope. The images are of superb quality, allowing the determination of radii, fluxes, and albedos. Attempts were made to improve the resolution of the already diffraction limited images by image restoration. These yielded indications of surface albedo distributions qualitatively consistent with models derived from observations of Pluto-Charon mutual eclipses.

  3. High resolution projection micro stereolithography system and method

    DOEpatents

    Spadaccini, Christopher M.; Farquar, George; Weisgraber, Todd; Gemberling, Steven; Fang, Nicholas; Xu, Jun; Alonso, Matthew; Lee, Howon

    2016-11-15

    A high-resolution P.mu.SL system and method incorporating one or more of the following features with a standard P.mu.SL system using a SLM projected digital image to form components in a stereolithographic bath: a far-field superlens for producing sub-diffraction-limited features, multiple spatial light modulators (SLM) to generate spatially-controlled three-dimensional interference holograms with nanoscale features, and the integration of microfluidic components into the resin bath of a P.mu.SL system to fabricate microstructures of different materials.

  4. Influence of seismic diffraction for high-resolution imaging: applications in offshore Malaysia

    NASA Astrophysics Data System (ADS)

    Bashir, Yasir; Ghosh, Deva Prasad; Sum, Chow Weng

    2018-04-01

    Small-scale geological discontinuities are not easy to detect and image in seismic data, as these features represent themselves as diffracted rather than reflected waves. However, the combined reflected and diffracted image contains full wave information and is of great value to an interpreter, for instance enabling the identification of faults, fractures, and surfaces in built-up carbonate. Although diffraction imaging has a resolution below the typical seismic wavelength, if the wavelength is much smaller than the width of the discontinuity then interference effects can be ignored, as they would not play a role in generating the seismic diffractions. In this paper, by means of synthetic examples and real data, the potential of diffraction separation for high-resolution seismic imaging is revealed and choosing the best method for preserving diffraction are discussed. We illustrate the accuracy of separating diffractions using the plane-wave destruction (PWD) and dip frequency filtering (DFF) techniques on data from the Sarawak Basin, a carbonate field. PWD is able to preserve the diffraction more intelligently than DFF, which is proven in the results by the model and real data. The final results illustrate the effectiveness of diffraction separation and possible imaging for high-resolution seismic data of small but significant geological features.

  5. Dual-color STED microscopy reveals a sandwich structure of Bassoon and Piccolo in active zones of adult and aged mice.

    PubMed

    Nishimune, Hiroshi; Badawi, Yomna; Mori, Shuuichi; Shigemoto, Kazuhiro

    2016-06-20

    Presynaptic active zones play a pivotal role as synaptic vesicle release sites for synaptic transmission, but the molecular architecture of active zones in mammalian neuromuscular junctions (NMJs) at sub-diffraction limited resolution remains unknown. Bassoon and Piccolo are active zone specific cytosolic proteins essential for active zone assembly in NMJs, ribbon synapses, and brain synapses. These proteins are thought to colocalize and share some functions at active zones. Here, we report an unexpected finding of non-overlapping localization of these two proteins in mouse NMJs revealed using dual-color stimulated emission depletion (STED) super resolution microscopy. Piccolo puncta sandwiched Bassoon puncta and aligned in a Piccolo-Bassoon-Piccolo structure in adult NMJs. P/Q-type voltage-gated calcium channel (VGCC) puncta colocalized with Bassoon puncta. The P/Q-type VGCC and Bassoon protein levels decreased significantly in NMJs from aged mouse. In contrast, the Piccolo levels in NMJs from aged mice were comparable to levels in adult mice. This study revealed the molecular architecture of active zones in mouse NMJs at sub-diffraction limited resolution, and described the selective degeneration mechanism of active zone proteins in NMJs from aged mice. Interestingly, the localization pattern of active zone proteins described herein is similar to active zone structures described using electron microscope tomography.

  6. Demonstration of nanoimprinted hyperlens array for high-throughput sub-diffraction imaging

    NASA Astrophysics Data System (ADS)

    Byun, Minsueop; Lee, Dasol; Kim, Minkyung; Kim, Yangdoo; Kim, Kwan; Ok, Jong G.; Rho, Junsuk; Lee, Heon

    2017-04-01

    Overcoming the resolution limit of conventional optics is regarded as the most important issue in optical imaging science and technology. Although hyperlenses, super-resolution imaging devices based on highly anisotropic dispersion relations that allow the access of high-wavevector components, have recently achieved far-field sub-diffraction imaging in real-time, the previously demonstrated devices have suffered from the extreme difficulties of both the fabrication process and the non-artificial objects placement. This results in restrictions on the practical applications of the hyperlens devices. While implementing large-scale hyperlens arrays in conventional microscopy is desirable to solve such issues, it has not been feasible to fabricate such large-scale hyperlens array with the previously used nanofabrication methods. Here, we suggest a scalable and reliable fabrication process of a large-scale hyperlens device based on direct pattern transfer techniques. We fabricate a 5 cm × 5 cm size hyperlenses array and experimentally demonstrate that it can resolve sub-diffraction features down to 160 nm under 410 nm wavelength visible light. The array-based hyperlens device will provide a simple solution for much more practical far-field and real-time super-resolution imaging which can be widely used in optics, biology, medical science, nanotechnology and other closely related interdisciplinary fields.

  7. Characterisation of phases in nanostructured, multilayered titanium alloys by analytical and high-resolution electron microscopy.

    PubMed

    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.

  8. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kiyota, Eduardo; Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas, Campinas-SP; Sousa, Sylvia Morais de

    Preliminary X-ray diffraction studies of apo maize aldose reductase at 2.0 Å resolution are reported. Maize aldose reductase (AR) is a member of the aldo-keto reductase superfamily. In contrast to human AR, maize AR seems to prefer the conversion of sorbitol into glucose. The apoenzyme was crystallized in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 47.2, b = 54.5, c = 100.6 Å and one molecule in the asymmetric unit. Synchrotron X-ray diffraction data were collected and a final resolution limit of 2.0 Å was obtained after data reduction. Phasing was carried out by an automatedmore » molecular-replacement procedure and structural refinement is currently in progress. The refined structure is expected to shed light on the functional/enzymatic mechanism and the unusual activities of maize AR.« less

  9. High-resolution mid-infrared observations of NGC 7469

    NASA Technical Reports Server (NTRS)

    Miles, J. W.; Houck, J. R.; Hayward, T. L.

    1994-01-01

    We present a high-resolution 11.7 micrometer image of the starburst/Seyfert hybrid galaxy NGC 7469 using the Hale 5 m telescope at Palomar Observatory. Our map, with diffraction limited spatial resolution of 0.6 sec, shows a 3 sec diameter ring of emission around an unresolved nucleus. The map is similar to the Very Large Array (VLA) 6 cm map of this galaxy made with 0.4 sec resolution by Wilson et al. (1991). About half of the mid-infrared flux in our map emerges from the unresolved nucleus. We also present spatially resolved low resolution spectra that show that the 11.3 micrometer polycyclic aromatic hydrocarbon (PAH) feature comes from the circumnuclear ring but not from the nucleus of the galaxy.

  10. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gao, Lan; Hill, K. W.; Bitter, M.

    Here, a high spatial resolution of a few μm is often required for probing small-scale high-energy-density plasmas using high resolution x-ray imaging spectroscopy. This resolution can be achieved by adjusting system magnification to overcome the inherent limitation of the detector pixel size. Laboratory experiments on investigating the relation between spatial resolution and system magnification for a spherical crystal spectrometer are presented. Tungsten Lβ 2 rays from a tungsten-target micro-focus x-ray tube were diffracted by a Ge 440 crystal, which was spherically bent to a radius of 223 mm, and imaged onto an x-ray CCD with 13-μm pixel size. The source-to-crystalmore » (p) and crystal-to-detector (q) distances were varied to produce spatial magnifications ( M = q/p) ranging from 2 to 10. The inferred instrumental spatial width reduces with increasing system magnification M. However, the experimental measurement at each M is larger than the theoretical value of pixel size divided by M. Future work will focus on investigating possible broadening mechanisms that limit the spatial resolution.« less

  11. Interlaced zone plate optics for hard X-ray imaging in the 10 nm range

    DOE PAGES

    Mohacsi, Istvan; Vartiainen, Ismo; Rosner, Benedikt; ...

    2017-03-08

    Multi-keV X-ray microscopy has been particularly successful in bridging the resolution gap between optical and electron microscopy. However, resolutions below 20 nm are still considered challenging, as high throughput direct imaging methods are limited by the availability of suitable optical elements. In order to bridge this gap, we present a new type of Fresnel zone plate lenses aimed at the sub-20 and the sub-10 nm resolution range. By extending the concept of double-sided zone plate stacking, we demonstrate the doubling of the effective line density and thus the resolution and provide large aperture, single- chip optical devices with 15 andmore » 7 nm smallest zone widths. The detailed characterization of these lenses shows excellent optical properties with focal spots down to 7.8 nm. Furthermore, beyond wave front characterization, the zone plates also excel in typical imaging scenarios, verifying their resolution close to their diffraction limited optical performance.« less

  12. Time multiplexing super-resolution nanoscopy based on the Brownian motion of gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Ilovitsh, Tali; Ilovitsh, Asaf; Wagner, Omer; Zalevsky, Zeev

    2017-02-01

    Super-resolution localization microscopy can overcome the diffraction limit and achieve a tens of order improvement in resolution. It requires labeling the sample with fluorescent probes followed with their repeated cycles of activation and photobleaching. This work presents an alternative approach that is free from direct labeling and does not require the activation and photobleaching cycles. Fluorescently labeled gold nanoparticles in a solution are distributed on top of the sample. The nanoparticles move in a random Brownian motion, and interact with the sample. By obscuring different areas in the sample, the nanoparticles encode the sub-wavelength features. A sequence of images of the sample is captured and decoded by digital post processing to create the super-resolution image. The achievable resolution is limited by the additive noise and the size of the nanoparticles. Regular nanoparticles with diameter smaller than 100nm are barely seen in a conventional bright field microscope, thus fluorescently labeled gold nanoparticles were used, with proper

  13. Interlaced zone plate optics for hard X-ray imaging in the 10 nm range

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mohacsi, Istvan; Vartiainen, Ismo; Rosner, Benedikt

    Multi-keV X-ray microscopy has been particularly successful in bridging the resolution gap between optical and electron microscopy. However, resolutions below 20 nm are still considered challenging, as high throughput direct imaging methods are limited by the availability of suitable optical elements. In order to bridge this gap, we present a new type of Fresnel zone plate lenses aimed at the sub-20 and the sub-10 nm resolution range. By extending the concept of double-sided zone plate stacking, we demonstrate the doubling of the effective line density and thus the resolution and provide large aperture, single- chip optical devices with 15 andmore » 7 nm smallest zone widths. The detailed characterization of these lenses shows excellent optical properties with focal spots down to 7.8 nm. Furthermore, beyond wave front characterization, the zone plates also excel in typical imaging scenarios, verifying their resolution close to their diffraction limited optical performance.« less

  14. Interlaced zone plate optics for hard X-ray imaging in the 10 nm range

    PubMed Central

    Mohacsi, Istvan; Vartiainen, Ismo; Rösner, Benedikt; Guizar-Sicairos, Manuel; Guzenko, Vitaliy A.; McNulty, Ian; Winarski, Robert; Holt, Martin V.; David, Christian

    2017-01-01

    Multi-keV X-ray microscopy has been particularly successful in bridging the resolution gap between optical and electron microscopy. However, resolutions below 20 nm are still considered challenging, as high throughput direct imaging methods are limited by the availability of suitable optical elements. In order to bridge this gap, we present a new type of Fresnel zone plate lenses aimed at the sub-20 and the sub-10 nm resolution range. By extending the concept of double-sided zone plate stacking, we demonstrate the doubling of the effective line density and thus the resolution and provide large aperture, singlechip optical devices with 15 and 7 nm smallest zone widths. The detailed characterization of these lenses shows excellent optical properties with focal spots down to 7.8 nm. Beyond wave front characterization, the zone plates also excel in typical imaging scenarios, verifying their resolution close to their diffraction limited optical performance.

  15. Interlaced zone plate optics for hard X-ray imaging in the 10 nm range

    NASA Astrophysics Data System (ADS)

    Mohacsi, Istvan; Vartiainen, Ismo; Rösner, Benedikt; Guizar-Sicairos, Manuel; Guzenko, Vitaliy A.; McNulty, Ian; Winarski, Robert; Holt, Martin V.; David, Christian

    2017-03-01

    Multi-keV X-ray microscopy has been particularly successful in bridging the resolution gap between optical and electron microscopy. However, resolutions below 20 nm are still considered challenging, as high throughput direct imaging methods are limited by the availability of suitable optical elements. In order to bridge this gap, we present a new type of Fresnel zone plate lenses aimed at the sub-20 and the sub-10 nm resolution range. By extending the concept of double-sided zone plate stacking, we demonstrate the doubling of the effective line density and thus the resolution and provide large aperture, singlechip optical devices with 15 and 7 nm smallest zone widths. The detailed characterization of these lenses shows excellent optical properties with focal spots down to 7.8 nm. Beyond wave front characterization, the zone plates also excel in typical imaging scenarios, verifying their resolution close to their diffraction limited optical performance.

  16. A super-resolution ultrasound method for brain vascular mapping

    PubMed Central

    O'Reilly, Meaghan A.; Hynynen, Kullervo

    2013-01-01

    Purpose: High-resolution vascular imaging has not been achieved in the brain due to limitations of current clinical imaging modalities. The authors present a method for transcranial ultrasound imaging of single micrometer-size bubbles within a tube phantom. Methods: Emissions from single bubbles within a tube phantom were mapped through an ex vivo human skull using a sparse hemispherical receiver array and a passive beamforming algorithm. Noninvasive phase and amplitude correction techniques were applied to compensate for the aberrating effects of the skull bone. The positions of the individual bubbles were estimated beyond the diffraction limit of ultrasound to produce a super-resolution image of the tube phantom, which was compared with microcomputed tomography (micro-CT). Results: The resulting super-resolution ultrasound image is comparable to results obtained via the micro-CT for small tissue specimen imaging. Conclusions: This method provides superior resolution to deep-tissue contrast ultrasound and has the potential to be extended to provide complete vascular network imaging in the brain. PMID:24320408

  17. Fast and efficient molecule detection in localization-based super-resolution microscopy by parallel adaptive histogram equalization.

    PubMed

    Li, Yiming; Ishitsuka, Yuji; Hedde, Per Niklas; Nienhaus, G Ulrich

    2013-06-25

    In localization-based super-resolution microscopy, individual fluorescent markers are stochastically photoactivated and subsequently localized within a series of camera frames, yielding a final image with a resolution far beyond the diffraction limit. Yet, before localization can be performed, the subregions within the frames where the individual molecules are present have to be identified-oftentimes in the presence of high background. In this work, we address the importance of reliable molecule identification for the quality of the final reconstructed super-resolution image. We present a fast and robust algorithm (a-livePALM) that vastly improves the molecule detection efficiency while minimizing false assignments that can lead to image artifacts.

  18. Wavelength scanning achieves pixel super-resolution in holographic on-chip microscopy

    NASA Astrophysics Data System (ADS)

    Luo, Wei; Göröcs, Zoltan; Zhang, Yibo; Feizi, Alborz; Greenbaum, Alon; Ozcan, Aydogan

    2016-03-01

    Lensfree holographic on-chip imaging is a potent solution for high-resolution and field-portable bright-field imaging over a wide field-of-view. Previous lensfree imaging approaches utilize a pixel super-resolution technique, which relies on sub-pixel lateral displacements between the lensfree diffraction patterns and the image sensor's pixel-array, to achieve sub-micron resolution under unit magnification using state-of-the-art CMOS imager chips, commonly used in e.g., mobile-phones. Here we report, for the first time, a wavelength scanning based pixel super-resolution technique in lensfree holographic imaging. We developed an iterative super-resolution algorithm, which generates high-resolution reconstructions of the specimen from low-resolution (i.e., under-sampled) diffraction patterns recorded at multiple wavelengths within a narrow spectral range (e.g., 10-30 nm). Compared with lateral shift-based pixel super-resolution, this wavelength scanning approach does not require any physical shifts in the imaging setup, and the resolution improvement is uniform in all directions across the sensor-array. Our wavelength scanning super-resolution approach can also be integrated with multi-height and/or multi-angle on-chip imaging techniques to obtain even higher resolution reconstructions. For example, using wavelength scanning together with multi-angle illumination, we achieved a halfpitch resolution of 250 nm, corresponding to a numerical aperture of 1. In addition to pixel super-resolution, the small scanning steps in wavelength also enable us to robustly unwrap phase, revealing the specimen's optical path length in our reconstructed images. We believe that this new wavelength scanning based pixel super-resolution approach can provide competitive microscopy solutions for high-resolution and field-portable imaging needs, potentially impacting tele-pathology applications in resource-limited-settings.

  19. High-resolution three-dimensional structural microscopy by single-angle Bragg ptychography

    DOE PAGES

    Hruszkewycz, S. O.; Allain, M.; Holt, M. V.; ...

    2016-11-21

    Coherent X-ray microscopy by phase retrieval of Bragg diffraction intensities enables lattice distortions within a crystal to be imaged at nanometre-scale spatial resolutions in three dimensions. While this capability can be used to resolve structure–property relationships at the nanoscale under working conditions, strict data measurement requirements can limit the application of current approaches. Here, in this work, we introduce an efficient method of imaging three-dimensional (3D) nanoscale lattice behaviour and strain fields in crystalline materials with a methodology that we call 3D Bragg projection ptychography (3DBPP). This method enables 3D image reconstruction of a crystal volume from a series ofmore » two-dimensional X-ray Bragg coherent intensity diffraction patterns measured at a single incident beam angle. Structural information about the sample is encoded along two reciprocal-space directions normal to the Bragg diffracted exit beam, and along the third dimension in real space by the scanning beam. Finally, we present our approach with an analytical derivation, a numerical demonstration, and an experimental reconstruction of lattice distortions in a component of a nanoelectronic prototype device.« less

  20. Merging single-shot XFEL diffraction data from inorganic nanoparticles: a new approach to size and orientation determination

    DOE PAGES

    Li, Xuanxuan; Spence, John C. H.; Hogue, Brenda G.; ...

    2017-09-22

    X-ray free-electron lasers (XFELs) provide new opportunities for structure determination of biomolecules, viruses and nanomaterials. With unprecedented peak brilliance and ultra-short pulse duration, XFELs can tolerate higher X-ray doses by exploiting the femtosecond-scale exposure time, and can thus go beyond the resolution limits achieved with conventional X-ray diffraction imaging techniques. Using XFELs, it is possible to collect scattering information from single particles at high resolution, however particle heterogeneity and unknown orientations complicate data merging in three-dimensional space. Using the Linac Coherent Light Source (LCLS), synthetic inorganic nanocrystals with a core–shell architecture were used as a model system for proof-of-principle coherentmore » diffractive single-particle imaging experiments. To deal with the heterogeneity of the core–shell particles, new computational methods have been developed to extract the particle size and orientation from the scattering data to assist data merging. The size distribution agrees with that obtained by electron microscopy and the merged data support a model with a core–shell architecture.« less

  1. Merging single-shot XFEL diffraction data from inorganic nanoparticles: a new approach to size and orientation determination

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, Xuanxuan; Spence, John C. H.; Hogue, Brenda G.

    X-ray free-electron lasers (XFELs) provide new opportunities for structure determination of biomolecules, viruses and nanomaterials. With unprecedented peak brilliance and ultra-short pulse duration, XFELs can tolerate higher X-ray doses by exploiting the femtosecond-scale exposure time, and can thus go beyond the resolution limits achieved with conventional X-ray diffraction imaging techniques. Using XFELs, it is possible to collect scattering information from single particles at high resolution, however particle heterogeneity and unknown orientations complicate data merging in three-dimensional space. Using the Linac Coherent Light Source (LCLS), synthetic inorganic nanocrystals with a core–shell architecture were used as a model system for proof-of-principle coherentmore » diffractive single-particle imaging experiments. To deal with the heterogeneity of the core–shell particles, new computational methods have been developed to extract the particle size and orientation from the scattering data to assist data merging. The size distribution agrees with that obtained by electron microscopy and the merged data support a model with a core–shell architecture.« less

  2. Femtosecond Electron Wave Packet Propagation and Diffraction: Towards Making the ``Molecular Movie"

    NASA Astrophysics Data System (ADS)

    Miller, R. J. Dwayne

    2003-03-01

    Time-resolved electron diffraction harbors great promise for achieving atomic resolution of the fastest chemical processes. The generation of sufficiently short electron pulses to achieve this real time view of a chemical reaction has been limited by problems in maintaining short electron pulses with realistic electron densities to the sample. The propagation dynamics of femtosecond electron packets in the drift region of a photoelectron gun are investigated with an N-body numerical simulation and mean-field model. This analyis shows that the redistribution of electrons inside the packet, arising from space-charge and dispersion contributions, changes the pulse envelope and leads to the development of a spatially linear axial velocity distribution. These results have been used in the design of femtosecond photoelectron guns with higher time resolution and novel electron-optical methods of pulse characterization that are approaching 100 fs timescales. Time-resolved diffraction studies with electron pulses of approximately 500 femtoseconds have focused on solid-liquid phase transitions under far from equilibrium conditions. This work gives a microscopic description of the melting process and illustrates the promise of atomically resolving transition state processes.

  3. Adaptive optics retinal imaging in the living mouse eye

    PubMed Central

    Geng, Ying; Dubra, Alfredo; Yin, Lu; Merigan, William H.; Sharma, Robin; Libby, Richard T.; Williams, David R.

    2012-01-01

    Correction of the eye’s monochromatic aberrations using adaptive optics (AO) can improve the resolution of in vivo mouse retinal images [Biss et al., Opt. Lett. 32(6), 659 (2007) and Alt et al., Proc. SPIE 7550, 755019 (2010)], but previous attempts have been limited by poor spot quality in the Shack-Hartmann wavefront sensor (SHWS). Recent advances in mouse eye wavefront sensing using an adjustable focus beacon with an annular beam profile have improved the wavefront sensor spot quality [Geng et al., Biomed. Opt. Express 2(4), 717 (2011)], and we have incorporated them into a fluorescence adaptive optics scanning laser ophthalmoscope (AOSLO). The performance of the instrument was tested on the living mouse eye, and images of multiple retinal structures, including the photoreceptor mosaic, nerve fiber bundles, fine capillaries and fluorescently labeled ganglion cells were obtained. The in vivo transverse and axial resolutions of the fluorescence channel of the AOSLO were estimated from the full width half maximum (FWHM) of the line and point spread functions (LSF and PSF), and were found to be better than 0.79 μm ± 0.03 μm (STD)(45% wider than the diffraction limit) and 10.8 μm ± 0.7 μm (STD)(two times the diffraction limit), respectively. The axial positional accuracy was estimated to be 0.36 μm. This resolution and positional accuracy has allowed us to classify many ganglion cell types, such as bistratified ganglion cells, in vivo. PMID:22574260

  4. Single particle tracking through highly scattering media with multiplexed two-photon excitation

    NASA Astrophysics Data System (ADS)

    Perillo, Evan; Liu, Yen-Liang; Liu, Cong; Yeh, Hsin-Chih; Dunn, Andrew K.

    2015-03-01

    3D single-particle tracking (SPT) has been a pivotal tool to furthering our understanding of dynamic cellular processes in complex biological systems, with a molecular localization accuracy (10-100 nm) often better than the diffraction limit of light. However, current SPT techniques utilize either CCDs or a confocal detection scheme which not only suffer from poor temporal resolution but also limit tracking to a depth less than one scattering mean free path in the sample (typically <15μm). In this report we highlight our novel design for a spatiotemporally multiplexed two-photon microscope which is able to reach sub-diffraction-limit tracking accuracy and sub-millisecond temporal resolution, but with a dramatically extended SPT range of up to 200 μm through dense cell samples. We have validated our microscope by tracking (1) fluorescent nanoparticles in a prescribed motion inside gelatin gel (with 1% intralipid) and (2) labeled single EGFR complexes inside skin cancer spheroids (at least 8 layers of cells thick) for ~10 minutes. Furthermore we discuss future capabilities of our multiplexed two-photon microscope design, specifically to the extension of (1) simultaneous multicolor tracking (i.e. spatiotemporal co-localization analysis) and (2) FRET studies (i.e. lifetime analysis). The high resolution, high depth penetration, and multicolor features of this microscope make it well poised to study a variety of molecular scale dynamics in the cell, especially related to cellular trafficking studies with in vitro tumor models and in vivo.

  5. Bringing the Visible Universe into Focus with Robo-AO

    PubMed Central

    Baranec, Christoph; Riddle, Reed; Law, Nicholas M.; Ramaprakash, A.N.; Tendulkar, Shriharsh P.; Bui, Khanh; Burse, Mahesh P.; Chordia, Pravin; Das, Hillol K.; Davis, Jack T.C.; Dekany, Richard G.; Kasliwal, Mansi M.; Kulkarni, Shrinivas R.; Morton, Timothy D.; Ofek, Eran O.; Punnadi, Sujit

    2013-01-01

    The angular resolution of ground-based optical telescopes is limited by the degrading effects of the turbulent atmosphere. In the absence of an atmosphere, the angular resolution of a typical telescope is limited only by diffraction, i.e., the wavelength of interest, λ, divided by the size of its primary mirror's aperture, D. For example, the Hubble Space Telescope (HST), with a 2.4-m primary mirror, has an angular resolution at visible wavelengths of ~0.04 arc seconds. The atmosphere is composed of air at slightly different temperatures, and therefore different indices of refraction, constantly mixing. Light waves are bent as they pass through the inhomogeneous atmosphere. When a telescope on the ground focuses these light waves, instantaneous images appear fragmented, changing as a function of time. As a result, long-exposure images acquired using ground-based telescopes - even telescopes with four times the diameter of HST - appear blurry and have an angular resolution of roughly 0.5 to 1.5 arc seconds at best. Astronomical adaptive-optics systems compensate for the effects of atmospheric turbulence. First, the shape of the incoming non-planar wave is determined using measurements of a nearby bright star by a wavefront sensor. Next, an element in the optical system, such as a deformable mirror, is commanded to correct the shape of the incoming light wave. Additional corrections are made at a rate sufficient to keep up with the dynamically changing atmosphere through which the telescope looks, ultimately producing diffraction-limited images. The fidelity of the wavefront sensor measurement is based upon how well the incoming light is spatially and temporally sampled1. Finer sampling requires brighter reference objects. While the brightest stars can serve as reference objects for imaging targets from several to tens of arc seconds away in the best conditions, most interesting astronomical targets do not have sufficiently bright stars nearby. One solution is to focus a high-power laser beam in the direction of the astronomical target to create an artificial reference of known shape, also known as a 'laser guide star'. The Robo-AO laser adaptive optics system2,3 employs a 10-W ultraviolet laser focused at a distance of 10 km to generate a laser guide star. Wavefront sensor measurements of the laser guide star drive the adaptive optics correction resulting in diffraction-limited images that have an angular resolution of ~0.1 arc seconds on a 1.5-m telescope. PMID:23426078

  6. Bringing the visible universe into focus with Robo-AO.

    PubMed

    Baranec, Christoph; Riddle, Reed; Law, Nicholas M; Ramaprakash, A N; Tendulkar, Shriharsh P; Bui, Khanh; Burse, Mahesh P; Chordia, Pravin; Das, Hillol K; Davis, Jack T C; Dekany, Richard G; Kasliwal, Mansi M; Kulkarni, Shrinivas R; Morton, Timothy D; Ofek, Eran O; Punnadi, Sujit

    2013-02-12

    The angular resolution of ground-based optical telescopes is limited by the degrading effects of the turbulent atmosphere. In the absence of an atmosphere, the angular resolution of a typical telescope is limited only by diffraction, i.e., the wavelength of interest, λ, divided by the size of its primary mirror's aperture, D. For example, the Hubble Space Telescope (HST), with a 2.4-m primary mirror, has an angular resolution at visible wavelengths of ~0.04 arc seconds. The atmosphere is composed of air at slightly different temperatures, and therefore different indices of refraction, constantly mixing. Light waves are bent as they pass through the inhomogeneous atmosphere. When a telescope on the ground focuses these light waves, instantaneous images appear fragmented, changing as a function of time. As a result, long-exposure images acquired using ground-based telescopes--even telescopes with four times the diameter of HST--appear blurry and have an angular resolution of roughly 0.5 to 1.5 arc seconds at best. Astronomical adaptive-optics systems compensate for the effects of atmospheric turbulence. First, the shape of the incoming non-planar wave is determined using measurements of a nearby bright star by a wavefront sensor. Next, an element in the optical system, such as a deformable mirror, is commanded to correct the shape of the incoming light wave. Additional corrections are made at a rate sufficient to keep up with the dynamically changing atmosphere through which the telescope looks, ultimately producing diffraction-limited images. The fidelity of the wavefront sensor measurement is based upon how well the incoming light is spatially and temporally sampled. Finer sampling requires brighter reference objects. While the brightest stars can serve as reference objects for imaging targets from several to tens of arc seconds away in the best conditions, most interesting astronomical targets do not have sufficiently bright stars nearby. One solution is to focus a high-power laser beam in the direction of the astronomical target to create an artificial reference of known shape, also known as a 'laser guide star'. The Robo-AO laser adaptive optics system, employs a 10-W ultraviolet laser focused at a distance of 10 km to generate a laser guide star. Wavefront sensor measurements of the laser guide star drive the adaptive optics correction resulting in diffraction-limited images that have an angular resolution of ~0.1 arc seconds on a 1.5-m telescope.

  7. Nanometres-resolution Kikuchi patterns from materials science specimens with transmission electron forward scatter diffraction in the scanning electron microscope.

    PubMed

    Brodusch, N; Demers, H; Gauvin, R

    2013-04-01

    A charge-coupled device camera of an electron backscattered diffraction system in a scanning electron microscope was positioned below a thin specimen and transmission Kikuchi patterns were collected. Contrary to electron backscattered diffraction, transmission electron forward scatter diffraction provides phase identification and orientation mapping at the nanoscale. The minimum Pd particle size for which a Kikuchi diffraction pattern was detected and indexed reliably was 5.6 nm. An orientation mapping resolution of 5 nm was measured at 30 kV. The resolution obtained with transmission electron forward scatter diffraction was of the same order of magnitude than that reported in electron nanodiffraction in the transmission electron microscope. An energy dispersive spectrometer X-ray map and a transmission electron forward scatter diffraction orientation map were acquired simultaneously. The high-resolution chemical, phase and orientation maps provided at once information on the chemical form, orientation and coherency of precipitates in an aluminium-lithium 2099 alloy. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

  8. Patterning via optical saturable transitions

    NASA Astrophysics Data System (ADS)

    Cantu, Precious

    For the past 40 years, optical lithography has been the patterning workhorse for the semiconductor industry. However, as integrated circuits have become more and more complex, and as device geometries shrink, more innovative methods are required to meet these needs. In the far-field, the smallest feature that can be generated with light is limited to approximately half the wavelength. This, so called far-field diffraction limit or the Abbe limit (after Prof. Ernst Abbe who first recognized this), effectively prevents the use of long-wavelength photons >300nm from patterning nanostructures <100nm. Even with a 193nm laser source and extremely complicated processing, patterns below ˜20nm are incredibly challenging to create. Sources with even shorter wavelengths can potentially be used. However, these tend be much more expensive and of much lower brightness, which in turn limits their patterning speed. Multi-photon reactions have been proposed to overcome the diffraction limit. However, these require very large intensities for modest gain in resolution. Moreover, the large intensities make it difficult to parallelize, thus limiting the patterning speed. In this dissertation, a novel nanopatterning technique using wavelength-selective small molecules that undergo single-photon reactions, enabling rapid top-down nanopatterning over large areas at low-light intensities, thereby allowing for the circumvention of the far-field diffraction barrier is developed and experimentally verified. This approach, which I refer to as Patterning via Optical Saturable Transitions (POST) has the potential for massive parallelism, enabling the creation of nanostructures and devices at a speed far surpassing what is currently possible with conventional optical lithographic techniques. The fundamental understanding of this technique goes beyond optical lithography in the semiconductor industry and is applicable to any area that requires the rapid patterning of large-area two or three-dimensional complex geometries. At a basic level, this research intertwines the fields of electrochemistry, material science, electrical engineering, optics, physics, and mechanical engineering with the goal of developing a novel super-resolution lithographic technique.

  9. Study on High Resolution Membrane-Based Diffractive Optical Imaging on Geostationary Orbit

    NASA Astrophysics Data System (ADS)

    Jiao, J.; Wang, B.; Wang, C.; Zhang, Y.; Jin, J.; Liu, Z.; Su, Y.; Ruan, N.

    2017-05-01

    Diffractive optical imaging technology provides a new way to realize high resolution earth observation on geostationary orbit. There are a lot of benefits to use the membrane-based diffractive optical element in ultra-large aperture optical imaging system, including loose tolerance, light weight, easy folding and unfolding, which make it easy to realize high resolution earth observation on geostationary orbit. The implementation of this technology also faces some challenges, including the configuration of the diffractive primary lens, the development of high diffraction efficiency membrane-based diffractive optical elements, and the correction of the chromatic aberration of the diffractive optical elements. Aiming at the configuration of the diffractive primary lens, the "6+1" petal-type unfold scheme is proposed, which consider the compression ratio, the blocking rate and the development complexity. For high diffraction efficiency membrane-based diffractive optical element, a self-collimating method is proposed. The diffraction efficiency is more than 90 % of the theoretical value. For the chromatic aberration correction problem, an optimization method based on schupmann is proposed to make the imaging spectral bandwidth in visible light band reach 100 nm. The above conclusions have reference significance for the development of ultra-large aperture diffractive optical imaging system.

  10. Resolution enhancement in coherent x-ray diffraction imaging by overcoming instrumental noise.

    PubMed

    Kim, Chan; Kim, Yoonhee; Song, Changyong; Kim, Sang Soo; Kim, Sunam; Kang, Hyon Chol; Hwu, Yeukuang; Tsuei, Ku-Ding; Liang, Keng San; Noh, Do Young

    2014-11-17

    We report that reference objects, strong scatterers neighboring weak phase objects, enhance the phase retrieval and spatial resolution in coherent x-ray diffraction imaging (CDI). A CDI experiment with Au nano-particles exhibited that the reference objects amplified the signal-to-noise ratio in the diffraction intensity at large diffraction angles, which significantly enhanced the image resolution. The interference between the diffracted x-ray from reference objects and a specimen also improved the retrieval of the phase of the diffraction signal. The enhancement was applied to image NiO nano-particles and a mitochondrion and confirmed in a simulation with a bacteria phantom. We expect that the proposed method will be of great help in imaging weakly scattering soft matters using coherent x-ray sources including x-ray free electron lasers.

  11. Optical resonance imaging: An optical analog to MRI with sub-diffraction-limited capabilities.

    PubMed

    Allodi, Marco A; Dahlberg, Peter D; Mazuski, Richard J; Davis, Hunter C; Otto, John P; Engel, Gregory S

    2016-12-21

    We propose here optical resonance imaging (ORI), a direct optical analog to magnetic resonance imaging (MRI). The proposed pulse sequence for ORI maps space to time and recovers an image from a heterodyne-detected third-order nonlinear photon echo measurement. As opposed to traditional photon echo measurements, the third pulse in the ORI pulse sequence has significant pulse-front tilt that acts as a temporal gradient. This gradient couples space to time by stimulating the emission of a photon echo signal from different lateral spatial locations of a sample at different times, providing a widefield ultrafast microscopy. We circumvent the diffraction limit of the optics by mapping the lateral spatial coordinate of the sample with the emission time of the signal, which can be measured to high precision using interferometric heterodyne detection. This technique is thus an optical analog of MRI, where magnetic-field gradients are used to localize the spin-echo emission to a point below the diffraction limit of the radio-frequency wave used. We calculate the expected ORI signal using 15 fs pulses and 87° of pulse-front tilt, collected using f /2 optics and find a two-point resolution 275 nm using 800 nm light that satisfies the Rayleigh criterion. We also derive a general equation for resolution in optical resonance imaging that indicates that there is a possibility of superresolution imaging using this technique. The photon echo sequence also enables spectroscopic determination of the input and output energy. The technique thus correlates the input energy with the final position and energy of the exciton.

  12. Mask-free, vacuum-free fabrication of high-conductivity metallic nanowire by spatially shaped ultrafast laser (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Wang, Andong; Li, Xiaowei; Qu, Lianti; Lu, Yongfeng; Jiang, Lan

    2017-03-01

    Metal nanowire fabrication has drawn tremendous attention in recent years due to its wide application in electronics, optoelectronics, and plasmonics. However, conventional laser fabrication technologies are limited by diffraction limit thus the fabrication resolution cannot meet the increasingly high demand of modern devices. Herein we report on a novel method for high-resolution high-quality metal nanowire fabrication by using Hermite-Gaussian beam to ablate metal thin film. The nanowire is formed due to the intensity valley in the center of the laser beam while the surrounding film is ablated. Arbitrary nanowire can be generated on the substrate by dynamically adjusting the orientation of the intensity valley. This method shows obvious advantages compared to conventional methods. First, the minimum nanowire has a width of 60 nm (≍1/13 of the laser wavelength), which is much smaller than the diffraction limit. The high resolution is achieved by combining the ultrashort nature of the femtosecond laser and the low thermal conductivity of the thin film. In addition, the fabricated nanowires have good inside qualities. No inner nanopores and particle intervals are generated inside the nanowire, thus endowing the nanowire with good electronic characteristics: the conductivity of the nanowires is as high as 1.2×107 S/m (≍1/4 of buck material), and the maximum current density is up to 1.66×108 A/m2. Last, the nanowire has a good adhesion to the substrates, which can withstand ultrasonic bath for a long time. These advantages make our method a good approach for high-resolution high-quality nanowire fabrication as a complementary method to conventional lithography methods.

  13. Ultrafast superresolution fluorescence imaging with spinning disk confocal microscope optics.

    PubMed

    Hayashi, Shinichi; Okada, Yasushi

    2015-05-01

    Most current superresolution (SR) microscope techniques surpass the diffraction limit at the expense of temporal resolution, compromising their applications to live-cell imaging. Here we describe a new SR fluorescence microscope based on confocal microscope optics, which we name the spinning disk superresolution microscope (SDSRM). Theoretically, the SDSRM is equivalent to a structured illumination microscope (SIM) and achieves a spatial resolution of 120 nm, double that of the diffraction limit of wide-field fluorescence microscopy. However, the SDSRM is 10 times faster than a conventional SIM because SR signals are recovered by optical demodulation through the stripe pattern of the disk. Therefore a single SR image requires only a single averaged image through the rotating disk. On the basis of this theory, we modified a commercial spinning disk confocal microscope. The improved resolution around 120 nm was confirmed with biological samples. The rapid dynamics of micro-tubules, mitochondria, lysosomes, and endosomes were observed with temporal resolutions of 30-100 frames/s. Because our method requires only small optical modifications, it will enable an easy upgrade from an existing spinning disk confocal to a SR microscope for live-cell imaging. © 2015 Hayashi and Okada. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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

    PubMed

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

    2010-04-22

    X-ray diffraction plays a pivotal role in the understanding of biological systems by revealing atomic structures of proteins, nucleic acids and their complexes, with much recent interest in very large assemblies like the ribosome. As crystals of such large assemblies often diffract weakly (resolution worse than 4 A), we need methods that work at such low resolution. In macromolecular assemblies, some of the components may be known at high resolution, whereas others are unknown: current refinement methods fail as they require a high-resolution starting structure for the entire complex. Determining the structure of such complexes, which are often of key biological importance, should be possible in principle as the number of independent diffraction intensities at a resolution better than 5 A generally exceeds the number of degrees of freedom. Here we introduce a method that adds specific information from known homologous structures but allows global and local deformations of these homology models. Our approach uses the observation that local protein structure tends to be conserved as sequence and function evolve. Cross-validation with R(free) (the free R-factor) determines the optimum deformation and influence of the homology model. For test cases at 3.5-5 A resolution with known structures at high resolution, our method gives significant improvements over conventional refinement in the model as monitored by coordinate accuracy, the definition of secondary structure and the quality of electron density maps. For re-refinements of a representative set of 19 low-resolution crystal structures from the Protein Data Bank, we find similar improvements. Thus, a structure derived from low-resolution diffraction data can have quality similar to a high-resolution structure. Our method is applicable to the study of weakly diffracting crystals using X-ray micro-diffraction as well as data from new X-ray light sources. Use of homology information is not restricted to X-ray crystallography and cryo-electron microscopy: as optical imaging advances to subnanometre resolution, it can use similar tools.

  15. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Huang X. R.; Siddons D.; Macrander, A.T.

    Realization of x-ray Fabry-Perot (FP) resonance in back-Bragg-reflection crystal cavities has been proposed and explored for many years, but to date no satisfactory performance has been achieved. Here we show that single-cavity crystal resonators intrinsically have limited finesse and efficiency. To break this limit, we demonstrate that monolithic multicavity resonators with equal-width cavities and specific plate thickness ratios can generate ultrahigh-resolution FP resonance with high efficiency, steep peak tails, and ultrahigh contrast simultaneously. The resonance mechanism is similar to that of sequentially cascaded single-cavity resonators. The ultranarrow-bandwidth FP resonance is anticipated to have various applications, including modern ultrahigh-resolution or precisionmore » x-ray monochromatization, spectroscopy, coherence purification, coherent diffraction, phase contrast imaging, etc.« less

  16. Cryogenic X-Ray Diffraction Microscopy for Biological Samples

    NASA Astrophysics Data System (ADS)

    Lima, Enju; Wiegart, Lutz; Pernot, Petra; Howells, Malcolm; Timmins, Joanna; Zontone, Federico; Madsen, Anders

    2009-11-01

    X-ray diffraction microscopy (XDM) is well suited for nondestructive, high-resolution biological imaging, especially for thick samples, with the high penetration power of x rays and without limitations imposed by a lens. We developed nonvacuum, cryogenic (cryo-) XDM with hard x rays at 8 keV and report the first frozen-hydrated imaging by XDM. By preserving samples in amorphous ice, the risk of artifacts associated with dehydration or chemical fixation is avoided, ensuring the imaging condition closest to their natural state. The reconstruction shows internal structures of intact D. radiodurans bacteria in their natural contrast.

  17. Third-dimension information retrieval from a single convergent-beam transmission electron diffraction pattern using an artificial neural network

    NASA Astrophysics Data System (ADS)

    Pennington, Robert S.; Van den Broek, Wouter; Koch, Christoph T.

    2014-05-01

    We have reconstructed third-dimension specimen information from convergent-beam electron diffraction (CBED) patterns simulated using the stacked-Bloch-wave method. By reformulating the stacked-Bloch-wave formalism as an artificial neural network and optimizing with resilient back propagation, we demonstrate specimen orientation reconstructions with depth resolutions down to 5 nm. To show our algorithm's ability to analyze realistic data, we also discuss and demonstrate our algorithm reconstructing from noisy data and using a limited number of CBED disks. Applicability of this reconstruction algorithm to other specimen parameters is discussed.

  18. Ultrasonic superlensing jets and acoustic-fork sheets

    NASA Astrophysics Data System (ADS)

    Mitri, F. G.

    2017-05-01

    Focusing acoustical (and optical) beams beyond the diffraction limit has remained a major challenge in imaging instruments and systems, until recent advances on ;hyper; or ;super; lensing and higher-resolution imaging techniques have shown the counterintuitive violation of this rule under certain circumstances. Nonetheless, the proposed technologies of super-resolution acoustical focusing beyond the diffraction barrier require complex tools such as artificially engineered metamaterials, and other hardware equipment that may not be easily synthesized or manufactured. The present contribution therefore suggests a simple and reliable method of using a sound-penetrable circular cylinder lens illuminated by a nonparaxial Gaussian acoustical sheet (i.e. finite beam in 2D) to produce non-evanescent ultrasonic superlensing jets (or bullets) and acoustical 'snail-fork' shaped wavefronts with limited diffraction. The generalized (near-field) scattering theory for acoustical sheets of arbitrary wavefronts and incidence is utilized to synthesize the incident beam based upon the angular spectrum decomposition method and the multipole expansion method in cylindrical wave functions to compute the scattered pressure around the cylinder with particular emphasis on its physical properties. The results show that depending on the beam and lens parameters, a tight focusing (with dimensions much smaller than the beam waist) can be achieved. Subwavelength resolution can be also achieved by selecting a lens material with a speed of sound exceeding that of the host fluid medium. The ultrasonic superlensing jets provide the impetus to develop improved subwavelength microscopy and acoustical image-slicing systems, cell lysis and surgery, and photoacoustic imaging to name a few examples. Moreover, an acoustical fork-sheet generation may open innovative avenues in reconfigurable on-chip micro/nanoparticle tweezers and surface acoustic waves devices.

  19. Coherent X-ray diffraction from collagenous soft tissues

    PubMed Central

    Berenguer de la Cuesta, Felisa; Wenger, Marco P. E.; Bean, Richard J.; Bozec, Laurent; Horton, Michael A.; Robinson, Ian K.

    2009-01-01

    Coherent X-ray diffraction has been applied in the imaging of inorganic materials with great success. However, its application to biological specimens has been limited to some notable exceptions, due to the induced radiation damage and the extended nature of biological samples, the last limiting the application of most part of the phasing algorithms. X-ray ptychography, still under development, is a good candidate to overcome such difficulties and become a powerful imaging method for biology. We describe herein the feasibility of applying ptychography to the imaging of biological specimens, in particular collagen rich samples. We report here speckles in diffraction patterns from soft animal tissue, obtained with an optimized small angle X-ray setup that exploits the natural coherence of the beam. By phasing these patterns, dark field images of collagen within tendon, skin, bone, or cornea will eventually be obtained with a resolution of 60–70 nm. We present simulations of the contrast mechanism in collagen based on atomic force microscope images of the samples. Simulations confirmed the ‘speckled’ nature of the obtained diffraction patterns. Once inverted, the patterns will show the disposition and orientation of the fibers within the tissue, by enhancing the phase contrast between protein and no protein regions of the sample. Our work affords the application of the most innovative coherent X-ray diffraction tools to the study of biological specimens, and this approach will have a significant impact in biology and medicine because it overcomes many of the limits of current microscopy techniques. PMID:19706395

  20. Coherent X-ray diffraction from collagenous soft tissues.

    PubMed

    Berenguer de la Cuesta, Felisa; Wenger, Marco P E; Bean, Richard J; Bozec, Laurent; Horton, Michael A; Robinson, Ian K

    2009-09-08

    Coherent X-ray diffraction has been applied in the imaging of inorganic materials with great success. However, its application to biological specimens has been limited to some notable exceptions, due to the induced radiation damage and the extended nature of biological samples, the last limiting the application of most part of the phasing algorithms. X-ray ptychography, still under development, is a good candidate to overcome such difficulties and become a powerful imaging method for biology. We describe herein the feasibility of applying ptychography to the imaging of biological specimens, in particular collagen rich samples. We report here speckles in diffraction patterns from soft animal tissue, obtained with an optimized small angle X-ray setup that exploits the natural coherence of the beam. By phasing these patterns, dark field images of collagen within tendon, skin, bone, or cornea will eventually be obtained with a resolution of 60-70 nm. We present simulations of the contrast mechanism in collagen based on atomic force microscope images of the samples. Simulations confirmed the 'speckled' nature of the obtained diffraction patterns. Once inverted, the patterns will show the disposition and orientation of the fibers within the tissue, by enhancing the phase contrast between protein and no protein regions of the sample. Our work affords the application of the most innovative coherent X-ray diffraction tools to the study of biological specimens, and this approach will have a significant impact in biology and medicine because it overcomes many of the limits of current microscopy techniques.

  1. Design and evaluation of a THz time domain imaging system using standard optical design software.

    PubMed

    Brückner, Claudia; Pradarutti, Boris; Müller, Ralf; Riehemann, Stefan; Notni, Gunther; Tünnermann, Andreas

    2008-09-20

    A terahertz (THz) time domain imaging system is analyzed and optimized with standard optical design software (ZEMAX). Special requirements to the illumination optics and imaging optics are presented. In the optimized system, off-axis parabolic mirrors and lenses are combined. The system has a numerical aperture of 0.4 and is diffraction limited for field points up to 4 mm and wavelengths down to 750 microm. ZEONEX is used as the lens material. Higher aspherical coefficients are used for correction of spherical aberration and reduction of lens thickness. The lenses were manufactured by ultraprecision machining. For optimization of the system, ray tracing and wave-optical methods were combined. We show how the ZEMAX Gaussian beam analysis tool can be used to evaluate illumination optics. The resolution of the THz system was tested with a wire and a slit target, line gratings of different period, and a Siemens star. The behavior of the temporal line spread function can be modeled with the polychromatic coherent line spread function feature in ZEMAX. The spectral and temporal resolutions of the line gratings are compared with the respective modulation transfer function of ZEMAX. For maximum resolution, the system has to be diffraction limited down to the smallest wavelength of the spectrum of the THz pulse. Then, the resolution on time domain analysis of the pulse maximum can be estimated with the spectral resolution of the center of gravity wavelength. The system resolution near the optical axis on time domain analysis of the pulse maximum is 1 line pair/mm with an intensity contrast of 0.22. The Siemens star is used for estimation of the resolution of the whole system. An eight channel electro-optic sampling system was used for detection. The resolution on time domain analysis of the pulse maximum of all eight channels could be determined with the Siemens star to be 0.7 line pairs/mm.

  2. Spatial resolution of a spherical x-ray crystal spectrometer at various magnifications

    DOE PAGES

    Gao, Lan; Hill, K. W.; Bitter, M.; ...

    2016-08-23

    Here, a high spatial resolution of a few μm is often required for probing small-scale high-energy-density plasmas using high resolution x-ray imaging spectroscopy. This resolution can be achieved by adjusting system magnification to overcome the inherent limitation of the detector pixel size. Laboratory experiments on investigating the relation between spatial resolution and system magnification for a spherical crystal spectrometer are presented. Tungsten Lβ 2 rays from a tungsten-target micro-focus x-ray tube were diffracted by a Ge 440 crystal, which was spherically bent to a radius of 223 mm, and imaged onto an x-ray CCD with 13-μm pixel size. The source-to-crystalmore » (p) and crystal-to-detector (q) distances were varied to produce spatial magnifications ( M = q/p) ranging from 2 to 10. The inferred instrumental spatial width reduces with increasing system magnification M. However, the experimental measurement at each M is larger than the theoretical value of pixel size divided by M. Future work will focus on investigating possible broadening mechanisms that limit the spatial resolution.« less

  3. Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM).

    PubMed

    Wang, Yilin; Kanchanawong, Pakorn

    2016-12-01

    Fluorescence microscopy enables direct visualization of specific biomolecules within cells. However, for conventional fluorescence microscopy, the spatial resolution is restricted by diffraction to ~ 200 nm within the image plane and > 500 nm along the optical axis. As a result, fluorescence microscopy has long been severely limited in the observation of ultrastructural features within cells. The recent development of super resolution microscopy methods has overcome this limitation. In particular, the advent of photoswitchable fluorophores enables localization-based super resolution microscopy, which provides resolving power approaching the molecular-length scale. Here, we describe the application of a three-dimensional super resolution microscopy method based on single-molecule localization microscopy and multiphase interferometry, called interferometric PhotoActivated Localization Microscopy (iPALM). This method provides nearly isotropic resolution on the order of 20 nm in all three dimensions. Protocols for visualizing the filamentous actin cytoskeleton, including specimen preparation and operation of the iPALM instrument, are described here. These protocols are also readily adaptable and instructive for the study of other ultrastructural features in cells.

  4. Three-dimensional wide-field pump-probe structured illumination microscopy

    PubMed Central

    Kim, Yang-Hyo; So, Peter T.C.

    2017-01-01

    We propose a new structured illumination scheme for achieving depth resolved wide-field pump-probe microscopy with sub-diffraction limit resolution. By acquiring coherent pump-probe images using a set of 3D structured light illumination patterns, a 3D super-resolution pump-probe image can be reconstructed. We derive the theoretical framework to describe the coherent image formation and reconstruction scheme for this structured illumination pump-probe imaging system and carry out numerical simulations to investigate its imaging performance. The results demonstrate a lateral resolution improvement by a factor of three and providing 0.5 µm level axial optical sectioning. PMID:28380860

  5. Fabrication and characterization of high-efficiency double-sided blazed x-ray optics.

    PubMed

    Mohacsi, Istvan; Vartiainen, Ismo; Guizar-Sicairos, Manuel; Karvinen, Petri; Guzenko, Vitaliy A; Müller, Elisabeth; Kewish, Cameron M; Somogyi, Andrea; David, Christian

    2016-01-15

    The focusing efficiency of conventional diffractive x-ray lenses is fundamentally limited due to their symmetric binary structures and the corresponding symmetry of their focusing and defocusing diffraction orders. Fresnel zone plates with asymmetric structure profiles can break this limitation; yet existing implementations compromise either on resolution, ease of use, or stability. We present a new way for the fabrication of such blazed lenses by patterning two complementary binary Fresnel zone plates on the front and back sides of the same membrane chip to provide a compact, inherently stable, single-chip device. The presented blazed double-sided zone plates with 200 nm smallest half-pitch provide up to 54.7% focusing efficiency at 6.2 keV, which is clearly beyond the value obtainable by their binary counterparts.

  6. Analysis of multi-channel microscopy: Spectral self-interference, multi-detector confocal and 4Pi systems

    NASA Astrophysics Data System (ADS)

    Davis, Brynmor J.

    Fluorescence microscopy is an important and ubiquitous tool in biological imaging due to the high specificity with which fluorescent molecules can be attached to an organism and the subsequent nondestructive in-vivo imaging allowed. Focused-light microscopies allow three-dimensional fluorescence imaging but their resolution is restricted by diffraction. This effect is particularly limiting in the axial dimension as the diffraction-limited focal volume produced by a lens is more extensive along the optical axis than perpendicular to it. Approaches such as confocal microscopy and 4Pi microscopy have been developed to improve the axial resolution. Spectral Self-Interference Fluorescence Microscopy (SSFM) is another high-axial-resolution technique and is the principal subject of this dissertation. Nanometer-precision localization of a single fluorescent layer has been demonstrated using SSFM. This accuracy compares favorably with the axial resolutions given by confocal and 4Pi systems at similar operating parameters (these resolutions are approximately 350nm and 80nm respectively). This theoretical work analyzes the expected performance of the SSFM system when imaging a general object, i.e. an arbitrary fluorophore density function rather than a single layer. An existing model of SSFM is used in simulations to characterize the system's resolution. Several statistically-based reconstruction methods are applied to show that the expected resolution for SSFM is similar to 4Pi microscopy for a general object but does give very high localization accuracy when the object is known to consist of a limited number of layers. SSFM is then analyzed in a linear systems framework and shown to have strong connections, both physically and mathematically, to a multi-channel 4Pi microscope. Fourier-domain analysis confirms that SSFM cannot be expected to outperform this multi-channel 4Pi instrument. Differences between the channels in spatial-scanning, multi-channel microscopies are then exploited to show that such instruments can operate at a sub-Nyquist scanning rate but still produce images largely free of aliasing effects. Multi-channel analysis is also used to show how light typically discarded in confocal and 4Pi systems can be collected and usefully incorporated into the measured image.

  7. Reconstruction from limited single-particle diffraction data via simultaneous determination of state, orientation, intensity, and phase

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Donatelli, Jeffrey J.; Sethian, James A.; Zwart, Peter H.

    Free-electron lasers now have the ability to collect X-ray diffraction patterns from individual molecules; however, each sample is delivered at unknown orientation and may be in one of several conformational states, each with a different molecular structure. Hit rates are often low, typically around 0.1%, limiting the number of useful images that can be collected. Determining accurate structural information requires classifying and orienting each image, accurately assembling them into a 3D diffraction intensity function, and determining missing phase information. Additionally, single particles typically scatter very few photons, leading to high image noise levels. We develop a multitiered iterative phasing algorithmmore » to reconstruct structural information from singleparticle diffraction data by simultaneously determining the states, orientations, intensities, phases, and underlying structure in a single iterative procedure. We leverage real-space constraints on the structure to help guide optimization and reconstruct underlying structure from very few images with excellent global convergence properties. We show that this approach can determine structural resolution beyond what is suggested by standard Shannon sampling arguments for ideal images and is also robust to noise.« less

  8. Reconstruction from limited single-particle diffraction data via simultaneous determination of state, orientation, intensity, and phase

    DOE PAGES

    Donatelli, Jeffrey J.; Sethian, James A.; Zwart, Peter H.

    2017-06-26

    Free-electron lasers now have the ability to collect X-ray diffraction patterns from individual molecules; however, each sample is delivered at unknown orientation and may be in one of several conformational states, each with a different molecular structure. Hit rates are often low, typically around 0.1%, limiting the number of useful images that can be collected. Determining accurate structural information requires classifying and orienting each image, accurately assembling them into a 3D diffraction intensity function, and determining missing phase information. Additionally, single particles typically scatter very few photons, leading to high image noise levels. We develop a multitiered iterative phasing algorithmmore » to reconstruct structural information from singleparticle diffraction data by simultaneously determining the states, orientations, intensities, phases, and underlying structure in a single iterative procedure. We leverage real-space constraints on the structure to help guide optimization and reconstruct underlying structure from very few images with excellent global convergence properties. We show that this approach can determine structural resolution beyond what is suggested by standard Shannon sampling arguments for ideal images and is also robust to noise.« less

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

    PubMed

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

    2018-06-01

    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.

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

    NASA Astrophysics Data System (ADS)

    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-invasive microscopy in animals and humans using ultrasound. We anticipate that ultrafast ultrasound localization microscopy may become an invaluable tool for the fundamental understanding and diagnostics of various disease processes that modify the microvascular blood flow, such as cancer, stroke and arteriosclerosis.

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

    PubMed

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

    2015-11-26

    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-invasive microscopy in animals and humans using ultrasound. We anticipate that ultrafast ultrasound localization microscopy may become an invaluable tool for the fundamental understanding and diagnostics of various disease processes that modify the microvascular blood flow, such as cancer, stroke and arteriosclerosis.

  12. Spider Silk: Mother Nature's Bio-Superlens

    NASA Astrophysics Data System (ADS)

    Monks, James N.; Yan, Bing; Hawkins, Nicholas; Vollrath, Fritz; Wang, Zengbo

    2016-09-01

    This paper demonstrates a possible new microfiber bio near field lens that uses minor ampullate spider silk,spun from the Nephila edulis spider, to create a real time image of a surface using near field optical techniques. The microfiber bio lens is the world's first natural superlens created by exploring biological materials. The resolution of the surface image overcomes the diffraction limit, with the ability to resolve patterns at 100 nm under a standard white light source in reflection mode. This resolution offers further developments in superlens technology and paves the way for new bio optics.

  13. Optical design for a breadboard high-resolution spectrometer for SIRTF/IRS

    NASA Astrophysics Data System (ADS)

    Brown, Robert J.; Houck, James R.; van Cleve, Jeffrey E.

    1996-11-01

    The optical design of a breadboard high resolution infrared spectrometer for the IRS instrument on the SIRTF mission is discussed. The spectrometer uses a crossed echelle grating configuration to cover the spectral region from 10 to 20 micrometer with a resolving power of approximately equals 600. The all reflective spectrometer forms a nearly diffraction limited image of the two dimensional spectrum on a 128 multiplied by 128 arsenic doped silicon area array with 75 micrometer pixels. The design aspects discussed include, grating numerology, image quality, packaging and alignment philosophy.

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

    NASA Astrophysics Data System (ADS)

    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.

  15. Solving the jitter problem in microwave compressed ultrafast electron diffraction instruments: Robust sub-50 fs cavity-laser phase stabilization

    PubMed Central

    Otto, M. R.; René de Cotret, L. P.; Stern, M. J.; Siwick, B. J.

    2017-01-01

    We demonstrate the compression of electron pulses in a high-brightness ultrafast electron diffraction instrument using phase-locked microwave signals directly generated from a mode-locked femtosecond oscillator. Additionally, a continuous-wave phase stabilization system that accurately corrects for phase fluctuations arising in the compression cavity from both power amplification and thermal drift induced detuning was designed and implemented. An improvement in the microwave timing stability from 100 fs to 5 fs RMS is measured electronically, and the long-term arrival time stability (>10 h) of the electron pulses improves to below our measurement resolution of 50 fs. These results demonstrate sub-relativistic ultrafast electron diffraction with compressed pulses that is no longer limited by laser-microwave synchronization. PMID:28852686

  16. Crystallization and preliminary X-ray diffraction studies of a novel ferredoxin involved in the dioxygenation of carbazole by Novosphingobium sp. KA1

    PubMed Central

    Umeda, Takashi; Katsuki, Junichi; Usami, Yusuke; Inoue, Kengo; Noguchi, Haruko; Fujimoto, Zui; Ashikawa, Yuji; Yamane, Hisakazu; Nojiri, Hideaki

    2008-01-01

    Novosphingobium sp. KA1 uses carbazole 1,9a-dioxygenase (CARDO) as the first dioxygenase in its carbazole-degradation pathway. The CARDO of KA1 contains a terminal oxygenase component and two electron-transfer components: ferredoxin and ferredoxin reductase. In contrast to the CARDO systems of other species, the ferredoxin component of KA1 is a putidaredoxin-type protein. This novel ferredoxin was crystallized at 293 K by the hanging-drop vapour-diffusion method using PEG MME 550 as the precipitant under anaerobic conditions. The crystals belong to space group C2221 and diffraction data were collected to a resolution of 1.9 Å (the diffraction limit was 1.6 Å). PMID:18607094

  17. Chromatic confocal microscope using hybrid aspheric diffractive lenses

    NASA Astrophysics Data System (ADS)

    Rayer, Mathieu; Mansfield, Daniel

    2014-05-01

    A chromatic confocal microscope is a single point non-contact distance measurement sensor. For three decades the vast majority of the chromatic confocal microscope use refractive-based lenses to code the measurement axis chromatically. However, such an approach is limiting the range of applications. In this paper the performance of refractive, diffractive and Hybrid aspheric diffractive are compared. Hybrid aspheric diffractive lenses combine the low geometric aberration of a diffractive lens with the high optical power of an aspheric lens. Hybrid aspheric diffractive lenses can reduce the number of elements in an imaging system significantly or create large hyper- chromatic lenses for sensing applications. In addition, diffractive lenses can improve the resolution and the dynamic range of a chromatic confocal microscope. However, to be suitable for commercial applications, the diffractive optical power must be significant. Therefore, manufacturing such lenses is a challenge. We show in this paper how a theoretical manufacturing model can demonstrate that the hybrid aspheric diffractive configuration with the best performances is achieved by step diffractive surface. The high optical quality of step diffractive surface is then demonstrated experimentally. Publisher's Note: This paper, originally published on 5/10/14, was replaced with a corrected/revised version on 5/19/14. If you downloaded the original PDF but are unable to access the revision, please contact SPIE Digital Library Customer Service for assistance.

  18. Holography and coherent diffraction with low-energy electrons: A route towards structural biology at the single molecule level.

    PubMed

    Latychevskaia, Tatiana; Longchamp, Jean-Nicolas; Escher, Conrad; Fink, Hans-Werner

    2015-12-01

    The current state of the art in structural biology is led by NMR, X-ray crystallography and TEM investigations. These powerful tools however all rely on averaging over a large ensemble of molecules. Here, we present an alternative concept aiming at structural analysis at the single molecule level. We show that by combining electron holography and coherent diffraction imaging estimations concerning the phase of the scattered wave become needless as the phase information is extracted from the data directly and unambiguously. Performed with low-energy electrons the resolution of this lens-less microscope is just limited by the De Broglie wavelength of the electron wave and the numerical aperture, given by detector geometry. In imaging freestanding graphene, a resolution of 2Å has been achieved revealing the 660.000 unit cells of the graphene sheet from a single data set. Once applied to individual biomolecules the method shall ultimately allow for non-destructive imaging and imports the potential to distinguish between different conformations of proteins with atomic resolution. Copyright © 2015. Published by Elsevier B.V.

  19. Three-dimensional refractive index and fluorescence tomography using structured illumination (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Park, GwangSik; Shin, SeungWoo; Kim, Kyoohyun; Park, YongKeun

    2017-02-01

    Optical diffraction tomography (ODT) has been an emerging optical technique for label-free imaging of three-dimensional (3-D) refractive index (RI) distribution of biological samples. ODT employs interferometric microscopy for measuring multiple holograms of samples with various incident angles, from which the Fourier diffraction theorem reconstructs the 3-D RI distribution of samples from retrieved complex optical fields. Since the RI value is linearly proportional to the protein concentration of biological samples where the proportional coefficient is called as refractive index increment (RII), reconstructed 3-D RI tomograms provide precise structural and biochemical information of individual biological samples. Because most proteins have similar RII value, however, ODT has limited molecular specificity, especially for imaging eukaryotic cells having various types of proteins and subcellular organelles. Here, we present an ODT system combined with structured illumination microscopy which can measure the 3-D RI distribution of biological samples as well as 3-D super-resolution fluorescent images in the same optical setup. A digital micromirror device (DMD) controls the incident angle of the illumination beam for tomogram reconstruction, and the same DMD modulates the structured illumination pattern of the excitation beam for super-resolution fluorescent imaging. We first validate the proposed method for simultaneous optical diffraction tomographic imaging and super-resolution fluorescent imaging of fluorescent beads. The proposed method is also exploited for various biological samples.

  20. A comprehensive simulation model of the performance of photochromic films in absorbance-modulation-optical-lithography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Majumder, Apratim; Helms, Phillip L.; Menon, Rajesh, E-mail: rmenon@eng.utah.edu

    2016-03-15

    Optical lithography is the most prevalent method of fabricating micro-and nano-scale structures in the semiconductor industry due to the fact that patterning using photons is fast, accurate and provides high throughput. However, the resolution of this technique is inherently limited by the physical phenomenon of diffraction. Absorbance-Modulation-Optical Lithography (AMOL), a recently developed technique has been successfully demonstrated to be able to circumvent this diffraction limit. AMOL employs a dual-wavelength exposure system in conjunction with spectrally selective reversible photo-transitions in thin films of photochromic molecules to achieve patterning of features with sizes beyond the far-field diffraction limit. We have developed amore » finite-element-method based full-electromagnetic-wave solution model that simulates the photo-chemical processes that occur within the thin film of the photochromic molecules under illumination by the exposure and confining wavelengths in AMOL. This model allows us to understand how the material characteristics influence the confinement to sub-diffraction dimensions, of the transmitted point spread function (PSF) of the exposure wavelength inside the recording medium. The model reported here provides the most comprehensive analysis of the AMOL process to-date, and the results show that the most important factors that govern the process, are the polarization of the two beams, the ratio of the intensities of the two wavelengths, the relative absorption coefficients and the concentration of the photochromic species, the thickness of the photochromic layer and the quantum yields of the photoreactions at the two wavelengths. The aim of this work is to elucidate the requirements of AMOL in successfully circumventing the far-field diffraction limit.« less

  1. High Resolution X-Ray Diffraction of Macromolecules with Synchrotron Radiation

    NASA Technical Reports Server (NTRS)

    Stojanoff, Vivian; Boggon, Titus; Helliwell, John R.; Judge, Russell; Olczak, Alex; Snell, Edward H.; Siddons, D. Peter; Rose, M. Franklin (Technical Monitor)

    2000-01-01

    We recently combined synchrotron-based monochromatic X-ray diffraction topography methods with triple axis diffractometry and rocking curve measurements: high resolution X-ray diffraction imaging techniques, to better understand the quality of protein crystals. We discuss these methods in the light of results obtained on crystals grown under different conditions. These non destructive techniques are powerful tools in the characterization of the protein crystals and ultimately will allow to improve, develop, and understand protein crystal growth. High resolution X-ray diffraction imaging methods will be discussed in detail in light of recent results obtained on Hen Egg White Lysozyme crystals and other proteins.

  2. High resolution imaging at Palomar

    NASA Technical Reports Server (NTRS)

    Kulkarni, Shrinivas R.

    1992-01-01

    For the last two years we have embarked on a program of understanding the ultimate limits of ground-based optical imaging. We have designed and fabricated a camera specifically for high resolution imaging. This camera has now been pressed into service at the prime focus of the Hale 5 m telescope. We have concentrated on two techniques: the Non-Redundant Masking (NRM) and Weigelt's Fully Filled Aperture (FFA) method. The former is the optical analog of radio interferometry and the latter is a higher order extension of the Labeyrie autocorrelation method. As in radio Very Long Baseline Interferometry (VLBI), both these techniques essentially measure the closure phase and, hence, true image construction is possible. We have successfully imaged binary stars and asteroids with angular resolution approaching the diffraction limit of the telescope and image quality approaching that of a typical radio VLBI map. In addition, we have carried out analytical and simulation studies to determine the ultimate limits of ground-based optical imaging, the limits of space-based interferometric imaging, and investigated the details of imaging tradeoffs of beam combination in optical interferometers.

  3. Imaging photonic crystals using hemispherical digital condensers and phase-recovery techniques.

    PubMed

    Alotaibi, Maged; Skinner-Ramos, Sueli; Farooq, Hira; Alharbi, Nouf; Alghasham, Hawra; de Peralta, Luis Grave

    2018-05-10

    We describe experiments where Fourier ptychographic microscopy (FPM) and dual-space microscopy (DSM) are implemented for imaging photonic crystals using a hemispherical digital condenser (HDC). Phase-recovery imaging simulations show that both techniques should be able to image photonic crystals with a period below the Rayleigh resolution limit. However, after processing the experimental images using both phase-recovery algorithms, we found that DSM can, but FPM cannot, image periodic structures with a period below the diffraction limit. We studied the origin of this apparent contradiction between simulations and experiments, and we concluded that the occurrence of unwanted reflections in the HDC is the source of the apparent failure of FPM. We thereafter solved the problem of reflections by using a single-directional illumination source and showed that FPM can image photonic crystals with a period below the Rayleigh resolution limit.

  4. Super-resolution photoacoustic microscopy using a localized near-field of a plasmonic nanoaperture: a three-dimensional simulation study

    NASA Astrophysics Data System (ADS)

    Park, Byullee; Lee, Hongki; Upputuri, Paul Kumar; Pramanik, Manojit; Kim, Donghyun; Kim, Chulhong

    2018-02-01

    Super-resolution microscopy has been increasingly important to delineate nanoscale biological structures or nanoparticles. With these increasing demands, several imaging modalities, including super-resolution fluorescence microscope (SRFM) and electron microscope (EM), have been developed and commercialized. These modalities achieve nanoscale resolution, however, SRFM cannot image without fluorescence, and sample preparation of EM is not suitable for biological specimens. To overcome those disadvantages, we have numerically studied the possibility of superresolution photoacoustic microscopy (SR-PAM) based on near-field localization of light. Photoacoustic (PA) signal is generally acquired based on optical absorption contrast; thus it requires no agents or pre-processing for the samples. The lateral resolution of the conventional photoacoustic microscopy is limited to 200 nm by diffraction limit, therefore reducing the lateral resolution is a major research impetus. Our approach to breaking resolution limit is to use laser pulses of extremely small spot size as a light source. In this research, we simulated the PA signal by constructing the three dimensional SR-PAM system environment using the k-Wave toolbox. As the light source, we simulated ultrashort light pulses using geometrical nanoaperture with near-field localization of surface plasmons. Through the PA simulation, we have successfully distinguish cuboids spaced 3 nm apart. In the near future, we will develop the SR-PAM and it will contribute to biomedical and material sciences.

  5. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gao, Lan, E-mail: lgao@pppl.gov; Hill, K. W.; Bitter, M.

    A high spatial resolution of a few μm is often required for probing small-scale high-energy-density plasmas using high resolution x-ray imaging spectroscopy. This resolution can be achieved by adjusting system magnification to overcome the inherent limitation of the detector pixel size. Laboratory experiments on investigating the relation between spatial resolution and system magnification for a spherical crystal spectrometer are presented. Tungsten Lβ{sub 2} rays from a tungsten-target micro-focus x-ray tube were diffracted by a Ge 440 crystal, which was spherically bent to a radius of 223 mm, and imaged onto an x-ray CCD with 13-μm pixel size. The source-to-crystal (p)more » and crystal-to-detector (q) distances were varied to produce spatial magnifications (M = q/p) ranging from 2 to 10. The inferred instrumental spatial width reduces with increasing system magnification M. However, the experimental measurement at each M is larger than the theoretical value of pixel size divided by M. Future work will focus on investigating possible broadening mechanisms that limit the spatial resolution.« less

  6. Recent advancements in structured-illumination microscopy toward live-cell imaging.

    PubMed

    Hirano, Yasuhiro; Matsuda, Atsushi; Hiraoka, Yasushi

    2015-08-01

    Fluorescence microscopy allows us to observe fluorescently labeled molecules in diverse biological processes and organelle structures within living cells. However, the diffraction limit restricts its spatial resolution to about half of its wavelength, limiting the capability of biological observation at the molecular level. Structured-illumination microscopy (SIM), a type of super-resolution microscopy, doubles the spatial resolution in all three dimensions by illuminating the sample with a patterned excitation light, followed by computer reconstruction. SIM uses a relatively low illumination power compared with other methods of super-resolution microscopy and is easily available for multicolor imaging. SIM has great potential for meeting the requirements of live-cell imaging. Recent developments in diverse types of SIM have achieved higher spatial (∼50 nm lateral) and temporal (∼100 Hz) resolutions. Here, we review recent advancements in SIM and discuss its application in noninvasive live-cell imaging. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Plasmon-Assisted Selective and Super-Resolving Excitation of Individual Quantum Emitters on a Metal Nanowire.

    PubMed

    Li, Qiang; Pan, Deng; Wei, Hong; Xu, Hongxing

    2018-03-14

    Hybrid systems composed of multiple quantum emitters coupled with plasmonic waveguides are promising building blocks for future integrated quantum nanophotonic circuits. The techniques that can super-resolve and selectively excite contiguous quantum emitters in a diffraction-limited area are of great importance for studying the plasmon-mediated interaction between quantum emitters and manipulating the single plasmon generation and propagation in plasmonic circuits. Here we show that multiple quantum dots coupled with a silver nanowire can be controllably excited by tuning the interference field of surface plasmons on the nanowire. Because of the period of the interference pattern is much smaller than the diffraction limit, we demonstrate the selective excitation of two quantum dots separated by a distance as short as 100 nm. We also numerically demonstrate a new kind of super-resolution imaging method that combines the tunable surface plasmon interference pattern on the NW with the structured illumination microscopy technique. Our work provides a novel high-resolution optical excitation and imaging method for the coupled systems of multiple quantum emitters and plasmonic waveguides, which adds a new tool for studying and manipulating single quantum emitters and single plasmons for quantum plasmonic circuitry applications.

  8. Binarization of apodizers by adapted one-dimensional error diffusion method

    NASA Astrophysics Data System (ADS)

    Kowalczyk, Marek; Cichocki, Tomasz; Martinez-Corral, Manuel; Andres, Pedro

    1994-10-01

    Two novel algorithms for the binarization of continuous rotationally symmetric real positive pupil filters are presented. Both algorithms are based on 1-D error diffusion concept. The original gray-tone apodizer is substituted by a set of transparent and opaque concentric annular zones. Depending on the algorithm the resulting binary mask consists of either equal width or equal area zones. The diffractive behavior of binary filters is evaluated. It is shown that the pupils with equal width zones give Fraunhofer diffraction pattern more similar to that of the original continuous-tone pupil than those with equal area zones, assuming in both cases the same resolution limit of printing device.

  9. Laser interferometry for the determination of thickness distributions of low absorbing films and their spatial and thickness resolutions.

    PubMed

    Mishima, T; Kao, K C

    1982-03-15

    New laser interferometry has been developed, based on the principle that a 2-D fringe pattern can be produced by interference of spatially coherent light beams. To avoid the effect of reflection from the back surface of the substrate, the Brewster angle of incidence is adopted; to suppress the effect of diffraction, a lens or a lens system is used. This laser interferometry is an efficient nondestructive technique for the determination of thickness distributions or uniformities of low absorbing films on transparent substrates over a large area without involving laborious computations. The limitation of spatial resolution, thickness resolution, and visibility of fringes is fully analyzed.

  10. Terahertz Near-Field Imaging Using Enhanced Transmission through a Single Subwavelength Aperture

    NASA Astrophysics Data System (ADS)

    Ishihara, Kunihiko; Ikari, Tomofumi; Minamide, Hiroaki; Shikata, Jun-ichi; Ohashi, Keishi; Yokoyama, Hiroyuki; Ito, Hiromasa

    2005-07-01

    We demonstrate terahertz (THz) near-field imaging using resonantly enhanced transmission of THz-wave radiation (λ˜ 200 μm) through a bull’s eye structure (a single subwavelength aperture surrounded by concentric periodic grooves in a metal plate). The bull’s eye structure shows extremely large enhanced transmission, which has the advantage for a single subwavelength aperture. The spatial resolution for the bull’s eye structure (with an aperture diameter d=100 μm) is evaluated in the near-field region, and a resolution of 50 μm (corresponding to λ/4) is achieved. We obtain the THz near-field images of the subwavelength metal pattern with a spatial resolution below the diffraction limit.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    Edrei, Eitan; Scarcelli, Giuliano

    2016-09-16

    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.

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

    NASA Astrophysics Data System (ADS)

    Grover, Ginni

    In recent years, super-resolution imaging has become an important fluorescent microscopy tool. It has enabled imaging of structures smaller than the optical diffraction limit with resolution less than 50 nm. Extension to high-resolution volume imaging has been achieved by integration with various optical techniques. In this thesis, development of a fluorescent microscope to enable high resolution, extended depth, three dimensional (3D) imaging is discussed; which is achieved by integration of computational methods with optical systems. In the first part of the thesis, point spread function (PSF) engineering for volume imaging is discussed. A class of PSFs, referred to as double-helix (DH) PSFs, is generated. The PSFs exhibit two focused spots in the image plane which rotate about the optical axis, encoding depth in rotation of the image. These PSFs extend the depth-of-field up to a factor of ˜5. Precision performance of the DH-PSFs, based on an information theoretical analysis, is compared with other 3D methods with conclusion that the DH-PSFs provide the best precision and the longest depth-of-field. Out of various possible DH-PSFs, a suitable PSF is obtained for super-resolution microscopy. The DH-PSFs are implemented in imaging systems, such as a microscope, with a special phase modulation at the pupil plane. Surface-relief elements which are polarization-insensitive and ˜90% light efficient are developed for phase modulation. The photon-efficient DH-PSF microscopes thus developed are used, along with optimal position estimation algorithms, for tracking and super-resolution imaging in 3D. Imaging at depths-of-field of up to 2.5 microm is achieved without focus scanning. Microtubules were imaged with 3D resolution of (6, 9, 39) nm, which is in close agreement with the theoretical limit. A quantitative study of co-localization of two proteins in volume was conducted in live bacteria. In the last part of the thesis practical aspects of the DH-PSF microscope are discussed. A method to stabilize it, for extended periods of time, with 3-4 nm precision in 3D is developed. 3D Super-resolution is demonstrated without drift. A PSF correction algorithm is demonstrated to improve characteristics of the DH-PSF in an experiment, where it is implemented with a polarization-insensitive liquid crystal spatial light modulator.

  14. 16 nm-resolution lithography using ultra-small-gap bowtie apertures

    NASA Astrophysics Data System (ADS)

    Chen, Yang; Qin, Jin; Chen, Jianfeng; Zhang, Liang; Ma, Chengfu; Chu, Jiaru; Xu, Xianfan; Wang, Liang

    2017-02-01

    Photolithography has long been a critical technology for nanoscale manufacturing, especially in the semiconductor industry. However, the diffractive nature of light has limited the continuous advance of optical lithography resolution. To overcome this obstacle, near-field scanning optical lithography (NSOL) is an alternative low-cost technique, whose resolution is determined by the near-field localization that can be achieved. Here, we apply the newly-developed backside milling method to fabricate bowtie apertures with a sub-15 nm gap, which can substantially improve the resolution of NSOL. A highly confined electric near field is produced by localized surface plasmon excitation and nanofocusing of the closely-tapered gap. We show contact lithography results with a record 16 nm resolution (FWHM). This photolithography scheme promises potential applications in data storage, high-speed computation, energy harvesting, and other nanotechnology areas.

  15. Nano-Optics for Chemical and Materials Characterization

    NASA Astrophysics Data System (ADS)

    Beversluis, Michael; Stranick, Stephan

    2007-03-01

    Light microscopy can provide non-destructive, real-time, three-dimensional imaging with chemically-specific contrast, but diffraction frequently limits the resolution to roughly 200 nm. Recently, structured illumination techniques have allowed fluorescence imaging to reach 50 nm resolution [1]. Since these fluorescence techniques were developed for use in microbiology, a key challenge is to take the resolution-enhancing features and apply them to contrast mechanisms like vibrational spectroscopy (e.g., Raman and CARS microscopy) that provide morphological and chemically specific imaging.. We are developing a new hybrid technique that combines the resolution enhancement of structured illumination microscopy with scanning techniques that can record hyperspectral images with 100 nm spatial resolution. We will show such superresolving images of semiconductor nanostructures and discuss the advantages and requirements for this technique. Referenence: 1. M. G. L. Gustafsson, P. Natl. Acad. Sci. USA 102, 13081-13086 (2005).

  16. Multi-pulse pumping for far-field super-resolution imaging

    NASA Astrophysics Data System (ADS)

    Requena, Sebastian; Raut, Sangram; Doan, Hung; Kimball, Joe; Fudala, Rafal; Borejdo, Julian; Gryczynski, Ignacy; Strzhemechny, Yuri; Gryczynski, Zygmunt

    2016-02-01

    Recently, far-field optical imaging with a resolution significantly beyond diffraction limit has attracted tremendous attention allowing for high resolution imaging in living objects. Various methods have been proposed that are divided in to two basic approaches; deterministic super-resolution like STED or RESOLFT and stochastic super-resolution like PALM or STORM. We propose to achieve super-resolution in far-field fluorescence imaging by the use of controllable (on-demand) bursts of pulses that can change the fluorescence signal of long-lived component over one order of magnitude. We demonstrate that two beads, one labeled with a long-lived dye and another with a short-lived dye, separated by a distance lower than 100 nm can be easily resolved in a single experiment. The proposed method can be used to separate two biological structures in a cell by targeting them with two antibodies labeled with long-lived and short-lived fluorophores.

  17. Single-particle coherent diffractive imaging with a soft x-ray free electron laser: towards soot aerosol morphology

    NASA Astrophysics Data System (ADS)

    Bogan, Michael J.; Starodub, Dmitri; Hampton, Christina Y.; Sierra, Raymond G.

    2010-10-01

    The first of its kind, the Free electron LASer facility in Hamburg, FLASH, produces soft x-ray pulses with unprecedented properties (10 fs, 6.8-47 nm, 1012 photons per pulse, 20 µm diameter). One of the seminal FLASH experiments is single-pulse coherent x-ray diffractive imaging (CXDI). CXDI utilizes the ultrafast and ultrabright pulses to overcome resolution limitations in x-ray microscopy imposed by x-ray-induced damage to the sample by 'diffracting before destroying' the sample on sub-picosecond timescales. For many lensless imaging algorithms used for CXDI it is convenient when the data satisfy an oversampling constraint that requires the sample to be an isolated object, i.e. an individual 'free-standing' portion of disordered matter delivered to the centre of the x-ray focus. By definition, this type of matter is an aerosol. This paper will describe the role of aerosol science methodologies used for the validation of the 'diffract before destroy' hypothesis and the execution of the first single-particle CXDI experiments being developed for biological imaging. FLASH CXDI now enables the highest resolution imaging of single micron-sized or smaller airborne particulate matter to date while preserving the native substrate-free state of the aerosol. Electron microscopy offers higher resolution for single-particle analysis but the aerosol must be captured on a substrate, potentially modifying the particle morphology. Thus, FLASH is poised to contribute significant advancements in our knowledge of aerosol morphology and dynamics. As an example, we simulate CXDI of combustion particle (soot) morphology and introduce the concept of extracting radius of gyration of fractal aggregates from single-pulse x-ray diffraction data. Future upgrades to FLASH will enable higher spatially and temporally resolved single-particle aerosol dynamics studies, filling a critical technological need in aerosol science and nanotechnology. Many of the methodologies described for FLASH will directly translate to use at hard x-ray free electron lasers.

  18. Digital direct electron imaging of energy-filtered electron backscatter diffraction patterns

    NASA Astrophysics Data System (ADS)

    Vespucci, S.; Winkelmann, A.; Naresh-Kumar, G.; Mingard, K. P.; Maneuski, D.; Edwards, P. R.; Day, A. P.; O'Shea, V.; Trager-Cowan, C.

    2015-11-01

    Electron backscatter diffraction is a scanning electron microscopy technique used to obtain crystallographic information on materials. It allows the nondestructive mapping of crystal structure, texture, and strain with a lateral and depth resolution on the order of tens of nanometers. Electron backscatter diffraction patterns (EBSPs) are presently acquired using a detector comprising a scintillator coupled to a digital camera, and the crystallographic information obtainable is limited by the conversion of electrons to photons and then back to electrons again. In this article we will report the direct acquisition of energy-filtered EBSPs using a digital complementary metal-oxide-semiconductor hybrid pixel detector, Timepix. We show results from a range of samples with different mass and density, namely diamond, silicon, and GaN. Direct electron detection allows the acquisition of EBSPs at lower (≤5 keV) electron beam energies. This results in a reduction in the depth and lateral extension of the volume of the specimen contributing to the pattern and will lead to a significant improvement in lateral and depth resolution. Direct electron detection together with energy filtering (electrons having energy below a specific value are excluded) also leads to an improvement in spatial resolution but in addition provides an unprecedented increase in the detail in the acquired EBSPs. An increase in contrast and higher-order diffraction features are observed. In addition, excess-deficiency effects appear to be suppressed on energy filtering. This allows the fundamental physics of pattern formation to be interrogated and will enable a step change in the use of electron backscatter diffraction (EBSD) for crystal phase identification and the mapping of strain. The enhancement in the contrast in high-pass energy-filtered EBSD patterns is found to be stronger for lighter, less dense materials. The improved contrast for such materials will enable the application of the EBSD technique to be expanded to materials for which conventional EBSD analysis is not presently practicable.

  19. Observations on the Role of Hydrogen in Facet Formation in Near-alpha Titanium (Preprint)

    DTIC Science & Technology

    2011-05-01

    using quantitative tilt fractography and electron backscatter diffraction while facet topography was examined using ultra high resolution scanning...quantitative tilt fractography and electron backscatter diffraction while facet topography was examined using ultra high resolution scanning electron...tilt fractography / electron backscatter diffraction (EBSD) technique in which both the crystallographic orientation of the fractured grain and the

  20. HYBRID Simulations of Diffraction-Limited Focusing with Kirkpatrick-Baez Mirrors for a Next-Generation In Situ Hard X-ray Nanoprobe

    NASA Astrophysics Data System (ADS)

    Maser, Jörg; Shi, Xianbo; Reininger, Ruben; Lai, Barry; Vogt, Stefan

    2016-12-01

    Next-generation hard X-ray nanoprobe beamlines such as the In Situ Nanoprobe (ISN) beamline being planned at the Advanced Photon Source aim at providing very high spatial resolution while also enabling very high focused flux, to study complex materials and devices using fast, multidimensional imaging across many length scales. The ISN will use diffractive optics to focus X-rays with a bandpass of Δ E/ E = 10-4 into a focal spot of 20 nm or below. Reflective optics in Kirkpatrick-Baez geometry will be used to focus X-rays with a bandpass as large as Δ E/ E = 10-2 into a focal spot of 50 nm. Diffraction-limited focusing with reflective optics is achieved by spatial filtering and use of a very long, vertically focusing mirror. To quantify the performance of the ISN beamline, we have simulated the propagation of both partially and fully coherent wavefronts from the undulator source, through the ISN beamline and into the mirror-based focal spot. Simulations were carried out using the recently developed software " HYBRID."

  1. Application of the phase extension method in virus crystallography.

    PubMed

    Reddy, Vijay S

    2016-01-01

    The procedure for phase extension (PX) involves gradually extending the initial phases from low resolution (e.g., ~8Å) to the high-resolution limit of a diffraction data set. Structural redundancy present in the viral capsids that display icosahedral symmetry results in a high degree of non-crystallographic symmetry (NCS), which in turn translates into higher phasing power and is critical for improving and extending phases to higher resolution. Greater completeness of the diffraction data and determination of a molecular replacement solution, which entails accurately identifying the virus particle orientation(s) and position(s), are important for the smooth progression of the PX procedure. In addition, proper definition of a molecular mask (envelope) around the NCS-asymmetric unit has been found to be important for the success of density modification procedures, such as density averaging and solvent flattening. Regardless of the degree of NCS, the PX method appears to work well in all space groups, provided an accurate molecular mask is used along with reasonable initial phases. However, in the cases with space group P1, in addition to requiring a molecular mask, starting the phase extension at a higher resolution (e.g., 6Å) overcame the previously reported problems due to Babinet phases and phase flipping errors.

  2. On effective and optical resolutions of diffraction data sets.

    PubMed

    Urzhumtseva, Ludmila; Klaholz, Bruno; Urzhumtsev, Alexandre

    2013-10-01

    In macromolecular X-ray crystallography, diffraction data sets are traditionally characterized by the highest resolution dhigh of the reflections that they contain. This measure is sensitive to individual reflections and does not refer to the eventual data incompleteness and anisotropy; it therefore does not describe the data well. A physically relevant and robust measure that provides a universal way to define the `actual' effective resolution deff of a data set is introduced. This measure is based on the accurate calculation of the minimum distance between two immobile point scatterers resolved as separate peaks in the Fourier map calculated with a given set of reflections. This measure is applicable to any data set, whether complete or incomplete. It also allows characterizion of the anisotropy of diffraction data sets in which deff strongly depends on the direction. Describing mathematical objects, the effective resolution deff characterizes the `geometry' of the set of measured reflections and is irrelevant to the diffraction intensities. At the same time, the diffraction intensities reflect the composition of the structure from physical entities: the atoms. The minimum distance for the atoms typical of a given structure is a measure that is different from and complementary to deff; it is also a characteristic that is complementary to conventional measures of the data-set quality. Following the previously introduced terms, this value is called the optical resolution, dopt. The optical resolution as defined here describes the separation of the atomic images in the `ideal' crystallographic Fourier map that would be calculated if the exact phases were known. The effective and optical resolution, as formally introduced in this work, are of general interest, giving a common `ruler' for all kinds of crystallographic diffraction data sets.

  3. Single-Molecule and Superresolution Imaging in Live Bacteria Cells

    PubMed Central

    Biteen, Julie S.; Moerner, W.E.

    2010-01-01

    Single-molecule imaging enables biophysical measurements devoid of ensemble averaging, gives enhanced spatial resolution beyond the diffraction limit, and permits superresolution reconstructions. Here, single-molecule and superresolution imaging are applied to the study of proteins in live Caulobacter crescentus cells to illustrate the power of these methods in bacterial imaging. Based on these techniques, the diffusion coefficient and dynamics of the histidine protein kinase PleC, the localization behavior of the polar protein PopZ, and the treadmilling behavior and protein superstructure of the structural protein MreB are investigated with sub-40-nm spatial resolution, all in live cells. PMID:20300204

  4. Some (Little) Thing(s) about VISIR

    NASA Astrophysics Data System (ADS)

    Pantin, E.; Vanzi, L.; Weilenmann, U.

    VISIR is the VLT mid-infrared Imager and Spectrometer. It offers a comprehensive set of observing modes, imaging in N and Q bands, at the limits of the telescope diffraction, as well as spectroscopy in the same bands. In particular, VISIR provides a very high-resolution spectroscopy mode with an achieved resolution up to 30000 in N band; this mode is so far unique in the southern hemisphere. VISIR calibration is quite specific when compared to standard visible/near-infrared ones. Various dedicated methods have to be developed to remove the instrumental signatures and obtain the best scientific return.

  5. Long working distance objective lenses for single atom trapping and imaging

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pritchard, J. D., E-mail: jonathan.pritchard@strath.ac.uk; Department of Physics, University of Strathclyde, 107 Rottenrow East, Glasgow G4 0NG; Isaacs, J. A.

    We present a pair of optimized objective lenses with long working distances of 117 mm and 65 mm, respectively, that offer diffraction limited performance for both Cs and Rb wavelengths when imaging through standard vacuum windows. The designs utilise standard catalog lens elements to provide a simple and cost-effective solution. Objective 1 provides NA = 0.175 offering 3 μm resolution whilst objective 2 is optimized for high collection efficiency with NA = 0.29 and 1.8 μm resolution. This flexible design can be further extended for use at shorter wavelengths by simply re-optimising the lens separations.

  6. Radiation damage free ghost diffraction with atomic resolution

    DOE PAGES

    Li, Zheng; Medvedev, Nikita; Chapman, Henry N.; ...

    2017-12-21

    The x-ray free electron lasers can enable diffractive structural determination of protein nanocrystals and single molecules that are too small and radiation-sensitive for conventional x-ray diffraction. However the electronic form factor may be modified during the ultrashort x-ray pulse due to photoionization and electron cascade caused by the intense x-ray pulse. For general x-ray imaging techniques, the minimization of the effects of radiation damage is of major concern to ensure reliable reconstruction of molecular structure. Here in this paper, we show that radiation damage free diffraction can be achieved with atomic spatial resolution by using x-ray parametric down-conversion and ghostmore » diffraction with entangled photons of x-ray and optical frequencies. We show that the formation of the diffraction patterns satisfies a condition analogous to the Bragg equation, with a resolution that can be as fine as the crystal lattice length scale of several Ångstrom. Since the samples are illuminated by low energy optical photons, they can be free of radiation damage.« less

  7. Radiation damage free ghost diffraction with atomic resolution

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, Zheng; Medvedev, Nikita; Chapman, Henry N.

    The x-ray free electron lasers can enable diffractive structural determination of protein nanocrystals and single molecules that are too small and radiation-sensitive for conventional x-ray diffraction. However the electronic form factor may be modified during the ultrashort x-ray pulse due to photoionization and electron cascade caused by the intense x-ray pulse. For general x-ray imaging techniques, the minimization of the effects of radiation damage is of major concern to ensure reliable reconstruction of molecular structure. Here in this paper, we show that radiation damage free diffraction can be achieved with atomic spatial resolution by using x-ray parametric down-conversion and ghostmore » diffraction with entangled photons of x-ray and optical frequencies. We show that the formation of the diffraction patterns satisfies a condition analogous to the Bragg equation, with a resolution that can be as fine as the crystal lattice length scale of several Ångstrom. Since the samples are illuminated by low energy optical photons, they can be free of radiation damage.« less

  8. Crystallization, dehydration and experimental phasing of WbdD, a bifunctional kinase and methyltransferase from Escherichia coli O9a

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hagelueken, Gregor; Huang, Hexian; Harlos, Karl

    2012-10-01

    The optimization of WbdD crystals using a novel dehydration protocol and experimental phasing at 3.5 Å resolution by cross-crystal averaging followed by molecular replacement of electron density into a non-isomorphous 3.0 Å resolution native data set are reported. WbdD is a bifunctional kinase/methyltransferase that is responsible for regulation of lipopolysaccharide O antigen polysaccharide chain length in Escherichia coli serotype O9a. Solving the crystal structure of this protein proved to be a challenge because the available crystals belonging to space group I23 only diffracted to low resolution (>95% of the crystals diffracted to resolution lower than 4 Å and most onlymore » to 8 Å) and were non-isomorphous, with changes in unit-cell dimensions of greater than 10%. Data from a serendipitously found single native crystal that diffracted to 3.0 Å resolution were non-isomorphous with a lower (3.5 Å) resolution selenomethionine data set. Here, a strategy for improving poor (3.5 Å resolution) initial phases by density modification and cross-crystal averaging with an additional 4.2 Å resolution data set to build a crude model of WbdD is desribed. Using this crude model as a mask to cut out the 3.5 Å resolution electron density yielded a successful molecular-replacement solution of the 3.0 Å resolution data set. The resulting map was used to build a complete model of WbdD. The hydration status of individual crystals appears to underpin the variable diffraction quality of WbdD crystals. After the initial structure had been solved, methods to control the hydration status of WbdD were developed and it was thus possible to routinely obtain high-resolution diffraction (to better than 2.5 Å resolution). This novel and facile crystal-dehydration protocol may be useful for similar challenging situations.« less

  9. DUV or EUV: that is the question

    NASA Astrophysics Data System (ADS)

    Williamson, David M.

    2000-11-01

    Lord Rayleigh's well-known equations for resolution and depth of focus indicate that resolution is better improved by reducing the wavelength of light rather than by increasing the numerical aperture (NA) of the projection optics, particularly when NA is approaching its physical limit of 1.0 in air (or vacuum). Vector aerial image simulations of diffraction-limited Deep Ultraviolet (DUV) and Extreme Ultraviolet (EUV) lithographic systems verify this simple view, even though Rayleigh's constants in Microlithography are not constant because of a variety of image enhancement techniques that attempt to compensate for the shortcomings of the aerial image when it is pushed to the limit. The aerial image is not the whole story, however. The competition between DUV and EUV systems will be decided more by economic and technological factors such as risk, time and cost of development and cost of ownership. These in turn depend on cost, availability and quality of light sources, refracting materials, photoresists and reticles.

  10. Single-pulse enhanced coherent diffraction imaging of bacteria with an X-ray free-electron laser

    NASA Astrophysics Data System (ADS)

    Fan, Jiadong; Sun, Zhibin; Wang, Yaling; Park, Jaehyun; Kim, Sunam; Gallagher-Jones, Marcus; Kim, Yoonhee; Song, Changyong; Yao, Shengkun; Zhang, Jian; Zhang, Jianhua; Duan, Xiulan; Tono, Kensuke; Yabashi, Makina; Ishikawa, Tetsuya; Fan, Chunhai; Zhao, Yuliang; Chai, Zhifang; Gao, Xueyun; Earnest, Thomas; Jiang, Huaidong

    2016-09-01

    High-resolution imaging offers one of the most promising approaches for exploring and understanding the structure and function of biomaterials and biological systems. X-ray free-electron lasers (XFELs) combined with coherent diffraction imaging can theoretically provide high-resolution spatial information regarding biological materials using a single XFEL pulse. Currently, the application of this method suffers from the low scattering cross-section of biomaterials and X-ray damage to the sample. However, XFELs can provide pulses of such short duration that the data can be collected using the “diffract and destroy” approach before the effects of radiation damage on the data become significant. These experiments combine the use of enhanced coherent diffraction imaging with single-shot XFEL radiation to investigate the cellular architecture of Staphylococcus aureus with and without labeling by gold (Au) nanoclusters. The resolution of the images reconstructed from these diffraction patterns were twice as high or more for gold-labeled samples, demonstrating that this enhancement method provides a promising approach for the high-resolution imaging of biomaterials and biological systems.

  11. Single-pulse enhanced coherent diffraction imaging of bacteria with an X-ray free-electron laser

    PubMed Central

    Fan, Jiadong; Sun, Zhibin; Wang, Yaling; Park, Jaehyun; Kim, Sunam; Gallagher-Jones, Marcus; Kim, Yoonhee; Song, Changyong; Yao, Shengkun; Zhang, Jian; Zhang, Jianhua; Duan, Xiulan; Tono, Kensuke; Yabashi, Makina; Ishikawa, Tetsuya; Fan, Chunhai; Zhao, Yuliang; Chai, Zhifang; Gao, Xueyun; Earnest, Thomas; Jiang, Huaidong

    2016-01-01

    High-resolution imaging offers one of the most promising approaches for exploring and understanding the structure and function of biomaterials and biological systems. X-ray free-electron lasers (XFELs) combined with coherent diffraction imaging can theoretically provide high-resolution spatial information regarding biological materials using a single XFEL pulse. Currently, the application of this method suffers from the low scattering cross-section of biomaterials and X-ray damage to the sample. However, XFELs can provide pulses of such short duration that the data can be collected using the “diffract and destroy” approach before the effects of radiation damage on the data become significant. These experiments combine the use of enhanced coherent diffraction imaging with single-shot XFEL radiation to investigate the cellular architecture of Staphylococcus aureus with and without labeling by gold (Au) nanoclusters. The resolution of the images reconstructed from these diffraction patterns were twice as high or more for gold-labeled samples, demonstrating that this enhancement method provides a promising approach for the high-resolution imaging of biomaterials and biological systems. PMID:27659203

  12. Single-pulse enhanced coherent diffraction imaging of bacteria with an X-ray free-electron laser.

    PubMed

    Fan, Jiadong; Sun, Zhibin; Wang, Yaling; Park, Jaehyun; Kim, Sunam; Gallagher-Jones, Marcus; Kim, Yoonhee; Song, Changyong; Yao, Shengkun; Zhang, Jian; Zhang, Jianhua; Duan, Xiulan; Tono, Kensuke; Yabashi, Makina; Ishikawa, Tetsuya; Fan, Chunhai; Zhao, Yuliang; Chai, Zhifang; Gao, Xueyun; Earnest, Thomas; Jiang, Huaidong

    2016-09-23

    High-resolution imaging offers one of the most promising approaches for exploring and understanding the structure and function of biomaterials and biological systems. X-ray free-electron lasers (XFELs) combined with coherent diffraction imaging can theoretically provide high-resolution spatial information regarding biological materials using a single XFEL pulse. Currently, the application of this method suffers from the low scattering cross-section of biomaterials and X-ray damage to the sample. However, XFELs can provide pulses of such short duration that the data can be collected using the "diffract and destroy" approach before the effects of radiation damage on the data become significant. These experiments combine the use of enhanced coherent diffraction imaging with single-shot XFEL radiation to investigate the cellular architecture of Staphylococcus aureus with and without labeling by gold (Au) nanoclusters. The resolution of the images reconstructed from these diffraction patterns were twice as high or more for gold-labeled samples, demonstrating that this enhancement method provides a promising approach for the high-resolution imaging of biomaterials and biological systems.

  13. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Berenguer de la Cuesta, Felisa; Wenger, Marco P.E.; Bean, Richard J.

    Coherent X-ray diffraction has been applied in the imaging of inorganic materials with great success. However, its application to biological specimens has been limited to some notable exceptions, due to the induced radiation damage and the extended nature of biological samples, the last limiting the application of most part of the phasing algorithms. X-ray ptychography, still under development, is a good candidate to overcome such difficulties and become a powerful imaging method for biology. We describe herein the feasibility of applying ptychography to the imaging of biological specimens, in particular collagen rich samples. We report here speckles in diffraction patternsmore » from soft animal tissue, obtained with an optimized small angle X-ray setup that exploits the natural coherence of the beam. By phasing these patterns, dark field images of collagen within tendon, skin, bone, or cornea will eventually be obtained with a resolution of 60-70 nm. We present simulations of the contrast mechanism in collagen based on atomic force microscope images of the samples. Simulations confirmed the 'speckled' nature of the obtained diffraction patterns. Once inverted, the patterns will show the disposition and orientation of the fibers within the tissue, by enhancing the phase contrast between protein and no protein regions of the sample. Our work affords the application of the most innovative coherent X-ray diffraction tools to the study of biological specimens, and this approach will have a significant impact in biology and medicine because it overcomes many of the limits of current microscopy techniques.« less

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

    PubMed Central

    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

  15. PtyNAMi: ptychographic nano-analytical microscope at PETRA III: interferometrically tracking positions for 3D x-ray scanning microscopy using a ball-lens retroreflector

    NASA Astrophysics Data System (ADS)

    Schroer, Christian G.; Seyrich, Martin; Kahnt, Maik; Botta, Stephan; Döhrmann, Ralph; Falkenberg, Gerald; Garrevoet, Jan; Lyubomirskiy, Mikhail; Scholz, Maria; Schropp, Andreas; Wittwer, Felix

    2017-09-01

    In recent years, ptychography has revolutionized x-ray microscopy in that it is able to overcome the diffraction limit of x-ray optics, pushing the spatial resolution limit down to a few nanometers. However, due to the weak interaction of x rays with matter, the detection of small features inside a sample requires a high coherent fluence on the sample, a high degree of mechanical stability, and a low background signal from the x-ray microscope. The x-ray scanning microscope PtyNAMi at PETRA III is designed for high-spatial-resolution 3D imaging with high sensitivity. The design concept is presented with a special focus on real-time metrology of the sample position during tomographic scanning microscopy.

  16. Near-isotropic 3D optical nanoscopy with photon-limited chromophores

    PubMed Central

    Tang, Jianyong; Akerboom, Jasper; Vaziri, Alipasha; Looger, Loren L.; Shank, Charles V.

    2010-01-01

    Imaging approaches based on single molecule localization break the diffraction barrier of conventional fluorescence microscopy, allowing for bioimaging with nanometer resolution. It remains a challenge, however, to precisely localize photon-limited single molecules in 3D. We have developed a new localization-based imaging technique achieving almost isotropic subdiffraction resolution in 3D. A tilted mirror is used to generate a side view in addition to the front view of activated single emitters, allowing their 3D localization to be precisely determined for superresolution imaging. Because both front and side views are in focus, this method is able to efficiently collect emitted photons. The technique is simple to implement on a commercial fluorescence microscope, and especially suitable for biological samples with photon-limited chromophores such as endogenously expressed photoactivatable fluorescent proteins. Moreover, this method is relatively resistant to optical aberration, as it requires only centroid determination for localization analysis. Here we demonstrate the application of this method to 3D imaging of bacterial protein distribution and neuron dendritic morphology with subdiffraction resolution. PMID:20472826

  17. Challenges in quantitative crystallographic characterization of 3D thin films by ACOM-TEM.

    PubMed

    Kobler, A; Kübel, C

    2017-02-01

    Automated crystal orientation mapping for transmission electron microscopy (ACOM-TEM) has become an easy to use method for the investigation of crystalline materials and complements other TEM methods by adding local crystallographic information over large areas. It fills the gap between high resolution electron microscopy and electron back scatter diffraction in terms of spatial resolution. Recent investigations showed that spot diffraction ACOM-TEM is a quantitative method with respect to sample parameters like grain size, twin density, orientation density and others. It can even be used in combination with in-situ tensile or thermal testing. However, there are limitations of the current method. In this paper we discuss some of the challenges and discuss solutions, e.g. we present an ambiguity filter that reduces the number of pixels with a '180° ambiguity problem'. For that an ACOM-TEM tilt series of nanocrystalline Pd thin films with overlapping crystallites was acquired and analyzed. Copyright © 2017. Published by Elsevier B.V.

  18. Biological applications of near-field scanning optical microscopy

    NASA Astrophysics Data System (ADS)

    Moers, Marco H. P.; Ruiter, A. G. T.; Jalocha, Alain; van Hulst, Niko F.; Kalle, W. H. J.; Wiegant, J. C. A. G.; Raap, A. K.

    1995-09-01

    Near-field Scanning Optical Microscopy (NSOM) is a true optical microscopic technique allowing fluorescence, absorption, reflection and polarization contrast with the additional advantage of nanometer lateral resolution, unlimited by diffraction and operation at ambient conditions. NSOM based on metal coated adiabatically tapered fibers, combined with shear force feedback and operated in illumination mode, has proven to be the most powerful NSOM arrangement, because of its true localization of the optical interaction, its various optical contrast possibilities and its sensitivity down to the single molecular level. In this paper applications of `aperture' NSOM to Fluorescence In Situ Hybridization of human metaphase chromosomes are presented, where the localized fluorescence allows to identify specific DNA sequences. All images are accompanied by the simultaneously acquired force image, enabling direct comparison of the optical contrast with the sample topography on nanometer scale, far beyond the diffraction limit. Thus the unique combination of high resolution, specific optical contrast and ambient operation offers many new direction possibilities in biological studies.

  19. Stochastic Optical Reconstruction Microscopy (STORM).

    PubMed

    Xu, Jianquan; Ma, Hongqiang; Liu, Yang

    2017-07-05

    Super-resolution (SR) fluorescence microscopy, a class of optical microscopy techniques at a spatial resolution below the diffraction limit, has revolutionized the way we study biology, as recognized by the Nobel Prize in Chemistry in 2014. Stochastic optical reconstruction microscopy (STORM), a widely used SR technique, is based on the principle of single molecule localization. STORM routinely achieves a spatial resolution of 20 to 30 nm, a ten-fold improvement compared to conventional optical microscopy. Among all SR techniques, STORM offers a high spatial resolution with simple optical instrumentation and standard organic fluorescent dyes, but it is also prone to image artifacts and degraded image resolution due to improper sample preparation or imaging conditions. It requires careful optimization of all three aspects-sample preparation, image acquisition, and image reconstruction-to ensure a high-quality STORM image, which will be extensively discussed in this unit. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  20. Fabrication and Operation of a Nano-Optical Conveyor Belt

    PubMed Central

    Ryan, Jason; Zheng, Yuxin; Hansen, Paul; Hesselink, Lambertus

    2015-01-01

    The technique of using focused laser beams to trap and exert forces on small particles has enabled many pivotal discoveries in the nanoscale biological and physical sciences over the past few decades. The progress made in this field invites further study of even smaller systems and at a larger scale, with tools that could be distributed more easily and made more widely available. Unfortunately, the fundamental laws of diffraction limit the minimum size of the focal spot of a laser beam, which makes particles smaller than a half-wavelength in diameter hard to trap and generally prevents an operator from discriminating between particles which are closer together than one half-wavelength. This precludes the optical manipulation of many closely-spaced nanoparticles and limits the resolution of optical-mechanical systems. Furthermore, manipulation using focused beams requires beam-forming or steering optics, which can be very bulky and expensive. To address these limitations in the system scalability of conventional optical trapping our lab has devised an alternative technique which utilizes near-field optics to move particles across a chip. Instead of focusing laser beams in the far-field, the optical near field of plasmonic resonators produces the necessary local optical intensity enhancement to overcome the restrictions of diffraction and manipulate particles at higher resolution. Closely-spaced resonators produce strong optical traps which can be addressed to mediate the hand-off of particles from one to the next in a conveyor-belt-like fashion. Here, we describe how to design and produce a conveyor belt using a gold surface patterned with plasmonic C-shaped resonators and how to operate it with polarized laser light to achieve super-resolution nanoparticle manipulation and transport. The nano-optical conveyor belt chip can be produced using lithography techniques and easily packaged and distributed. PMID:26381708

  1. Fabrication and Operation of a Nano-Optical Conveyor Belt.

    PubMed

    Ryan, Jason; Zheng, Yuxin; Hansen, Paul; Hesselink, Lambertus

    2015-08-26

    The technique of using focused laser beams to trap and exert forces on small particles has enabled many pivotal discoveries in the nanoscale biological and physical sciences over the past few decades. The progress made in this field invites further study of even smaller systems and at a larger scale, with tools that could be distributed more easily and made more widely available. Unfortunately, the fundamental laws of diffraction limit the minimum size of the focal spot of a laser beam, which makes particles smaller than a half-wavelength in diameter hard to trap and generally prevents an operator from discriminating between particles which are closer together than one half-wavelength. This precludes the optical manipulation of many closely-spaced nanoparticles and limits the resolution of optical-mechanical systems. Furthermore, manipulation using focused beams requires beam-forming or steering optics, which can be very bulky and expensive. To address these limitations in the system scalability of conventional optical trapping our lab has devised an alternative technique which utilizes near-field optics to move particles across a chip. Instead of focusing laser beams in the far-field, the optical near field of plasmonic resonators produces the necessary local optical intensity enhancement to overcome the restrictions of diffraction and manipulate particles at higher resolution. Closely-spaced resonators produce strong optical traps which can be addressed to mediate the hand-off of particles from one to the next in a conveyor-belt-like fashion. Here, we describe how to design and produce a conveyor belt using a gold surface patterned with plasmonic C-shaped resonators and how to operate it with polarized laser light to achieve super-resolution nanoparticle manipulation and transport. The nano-optical conveyor belt chip can be produced using lithography techniques and easily packaged and distributed.

  2. Intensity Interferometry: Imaging Stars with Kilometer Baselines

    NASA Astrophysics Data System (ADS)

    Dravins, Dainis

    2018-04-01

    Microarcsecond imaging will reveal stellar surfaces but requires kilometer-scale interferometers. Intensity interferometry circumvents atmospheric turbulence by correlating intensity fluctuations between independent telescopes. Telescopes connect only electronically, and the error budget relates to electronic timescales of nanoseconds (light-travel distances on the order of a meter), enabling the use of imperfect optics in a turbulent atmosphere. Once pioneered by Hanbury Brown and Twiss, digital versions have now been demonstrated in the laboratory, reconstructing diffraction-limited images from hundreds of optical baselines. Arrays of Cherenkov telescopes (primarily erected for gamma-ray studies) will extend over a few km, enabling an optical equivalent of radio interferometers. Resolutions in the tens of microarcseconds will resolve rotationally flattened stars with their circumstellar disks and winds, or possibly even the silhouettes of transiting exoplanets. Applying the method to mirror segments in extremely large telescopes (even with an incompletely filled main mirror, poor seeing, no adaptive optics), the diffraction limit in the blue may be reached.

  3. Scalable ion-photon quantum interface based on integrated diffractive mirrors

    NASA Astrophysics Data System (ADS)

    Ghadimi, Moji; Blūms, Valdis; Norton, Benjamin G.; Fisher, Paul M.; Connell, Steven C.; Amini, Jason M.; Volin, Curtis; Hayden, Harley; Pai, Chien-Shing; Kielpinski, David; Lobino, Mirko; Streed, Erik W.

    2017-12-01

    Quantum networking links quantum processors through remote entanglement for distributed quantum information processing and secure long-range communication. Trapped ions are a leading quantum information processing platform, having demonstrated universal small-scale processors and roadmaps for large-scale implementation. Overall rates of ion-photon entanglement generation, essential for remote trapped ion entanglement, are limited by coupling efficiency into single mode fibers and scaling to many ions. Here, we show a microfabricated trap with integrated diffractive mirrors that couples 4.1(6)% of the fluorescence from a 174Yb+ ion into a single mode fiber, nearly triple the demonstrated bulk optics efficiency. The integrated optic collects 5.8(8)% of the π transition fluorescence, images the ion with sub-wavelength resolution, and couples 71(5)% of the collected light into the fiber. Our technology is suitable for entangling multiple ions in parallel and overcomes mode quality limitations of existing integrated optical interconnects.

  4. Predicting scattering scanning near-field optical microscopy of mass-produced plasmonic devices

    NASA Astrophysics Data System (ADS)

    Otto, Lauren M.; Burgos, Stanley P.; Staffaroni, Matteo; Ren, Shen; Süzer, Özgün; Stipe, Barry C.; Ashby, Paul D.; Hammack, Aeron T.

    2018-05-01

    Scattering scanning near-field optical microscopy enables optical imaging and characterization of plasmonic devices with nanometer-scale resolution well below the diffraction limit. This technique enables developers to probe and understand the waveguide-coupled plasmonic antenna in as-fabricated heat-assisted magnetic recording heads. In order to validate and predict results and to extract information from experimental measurements that is physically comparable to simulations, a model was developed to translate the simulated electric field into expected near-field measurements using physical parameters specific to scattering scanning near-field optical microscopy physics. The methods used in this paper prove that scattering scanning near-field optical microscopy can be used to determine critical sub-diffraction-limited dimensions of optical field confinement, which is a crucial metrology requirement for the future of nano-optics, semiconductor photonic devices, and biological sensing where the near-field character of light is fundamental to device operation.

  5. Polarization-Dependent Quasi-Far-Field Superfocusing Strategy of Nanoring-Based Plasmonic Lenses.

    PubMed

    Sun, Hao; Zhu, Yechuan; Gao, Bo; Wang, Ping; Yu, Yiting

    2017-12-01

    The two-dimensional superfocusing of nanoring-based plasmonic lenses (NRPLs) beyond the diffraction limit in the far-field region remains a great challenge at optical wavelengths. In this paper, in addition to the modulation of structural parameters, we investigated the polarization-dependent focusing performance of a NRPL employing the finite-difference time-domain (FDTD) method. By utilizing the state of polarization (SOP) of incident light, we successfully realize the elliptical-, donut-, and circular-shape foci. The minimum full widths at half maximum (FWHMs) of these foci are ~0.32, ~0.34, and ~0.42 λ 0 in the total electric field, respectively, and the depth of focus (DOF) lies in 1.41~1.77 λ 0 . These sub-diffraction-limit foci are well controlled in the quasi-far-field region. The underlying physical mechanism on the focal shift and an effective way to control the focusing position are proposed. Furthermore, in the case of a high numerical aperture, the longitudinal component, which occupies over 80% of the electric-field energy, decides the focusing patterns of the foci. The achieved sub-diffraction-limit focusing can be widely used for many engineering applications, including the super-resolution imaging, particle acceleration, quantum optical information processing, and optical data storage.

  6. Femtosecond mega-electron-volt electron microdiffraction

    DOE PAGES

    Shen, X.; Li, R. K.; Lundstrom, U.; ...

    2017-09-01

    To understand and control the basic functions of physical, chemical and biological processes from micron to nano-meter scale, an instrument capable of visualizing transient structural changes of inhomogeneous materials with atomic spatial and temporal resolutions, is required. One such technique is femtosecond electron microdiffraction, in which a short electron pulse with femtosecond-scale duration is focused into a micron-scale spot and used to obtain diffraction images to resolve ultrafast structural dynamics over a localized crystalline domain. In this letter, we report the experimental demonstration of time-resolved mega-electron-volt electron microdiffraction which achieves a 5 μm root-mean-square (rms) beam size on the samplemore » and a 110 fs rms temporal resolution. Using pulses of 10k electrons at 4.2 MeV energy with a normalized emittance 3 nm-rad, we obtained high quality diffraction from a single 10 μm paraffin ( C 44 H 90) crystal. The phonon softening mode in optical-pumped polycrystalline Bi was also time-resolved, demonstrating the temporal resolution limits of the instrument. In conclusion, this new characterization capability will open many research opportunities in material and biological sciences.« less

  7. Femtosecond mega-electron-volt electron microdiffraction

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shen, X.; Li, R. K.; Lundstrom, U.

    To understand and control the basic functions of physical, chemical and biological processes from micron to nano-meter scale, an instrument capable of visualizing transient structural changes of inhomogeneous materials with atomic spatial and temporal resolutions, is required. One such technique is femtosecond electron microdiffraction, in which a short electron pulse with femtosecond-scale duration is focused into a micron-scale spot and used to obtain diffraction images to resolve ultrafast structural dynamics over a localized crystalline domain. In this letter, we report the experimental demonstration of time-resolved mega-electron-volt electron microdiffraction which achieves a 5 μm root-mean-square (rms) beam size on the samplemore » and a 110 fs rms temporal resolution. Using pulses of 10k electrons at 4.2 MeV energy with a normalized emittance 3 nm-rad, we obtained high quality diffraction from a single 10 μm paraffin ( C 44 H 90) crystal. The phonon softening mode in optical-pumped polycrystalline Bi was also time-resolved, demonstrating the temporal resolution limits of the instrument. In conclusion, this new characterization capability will open many research opportunities in material and biological sciences.« less

  8. Real-time and sub-wavelength ultrafast coherent diffraction imaging in the extreme ultraviolet.

    PubMed

    Zürch, M; Rothhardt, J; Hädrich, S; Demmler, S; Krebs, M; Limpert, J; Tünnermann, A; Guggenmos, A; Kleineberg, U; Spielmann, C

    2014-12-08

    Coherent Diffraction Imaging is a technique to study matter with nanometer-scale spatial resolution based on coherent illumination of the sample with hard X-ray, soft X-ray or extreme ultraviolet light delivered from synchrotrons or more recently X-ray Free-Electron Lasers. This robust technique simultaneously allows quantitative amplitude and phase contrast imaging. Laser-driven high harmonic generation XUV-sources allow table-top realizations. However, the low conversion efficiency of lab-based sources imposes either a large scale laser system or long exposure times, preventing many applications. Here we present a lensless imaging experiment combining a high numerical aperture (NA = 0.8) setup with a high average power fibre laser driven high harmonic source. The high flux and narrow-band harmonic line at 33.2 nm enables either sub-wavelength spatial resolution close to the Abbe limit (Δr = 0.8λ) for long exposure time, or sub-70 nm imaging in less than one second. The unprecedented high spatial resolution, compactness of the setup together with the real-time capability paves the way for a plethora of applications in fundamental and life sciences.

  9. The Development of a Scanning Soft X-Ray Microscope.

    NASA Astrophysics Data System (ADS)

    Rarback, Harvey Miles

    We have developed a scanning soft X-ray microscope, which can be used to image natural biological specimens at high resolution and with less damage than electron microscopy. The microscope focuses a monochromatic beam of synchrotron radiation to a nearly diffraction limited spot with the aid of a high resolution Fresnel zone plate, specially fabricated for us at the IBM Watson Research Center. The specimen at one atmosphere is mechanically scanned through the spot and the transmitted radiation is efficiently detected with a flow proportional counter. A computer forms a realtime transmission image of the specimen which is displayed on a color monitor. Our first generation optics have produced images of natural wet specimens at a resolution of 300 nm.

  10. Nanopositioning for polarimetric characterization.

    PubMed

    Qureshi, Naser; Kolokoltsev, Oleg V; Ortega-Martínez, Roberto; Ordoñez-Romero, C L

    2008-12-01

    A positioning system with approximately nanometer resolution has been developed based on a new implementation of a motor-driven screw scheme. In contrast to conventional positioning systems based on piezoelectric elements, this system shows remarkably low levels of drift and vibration, and eliminates the need for position feedback during typical data acquisition processes. During positioning or scanning processes, non-repeatability and hysteresis problems inherent in mechanical positioning systems are greatly reduced using a software feedback scheme. As a result, we are able to demonstrate an average mechanical resolution of 1.45 nm and near diffraction-limited imaging using scanning optical microscopy. We propose this approach to nanopositioning as a readily accessible alternative enabling high spatial resolution scanning probe characterization (e.g., polarimetry) and provide practical details for its implementation.

  11. Depth Resolution Dependence on Sample Thickness and Incident Energy in On-Axis Transmission Kikuchi Diffraction in Scanning Electron Microscope (SEM).

    PubMed

    Brodu, Etienne; Bouzy, Emmanuel

    2017-12-01

    Transmission Kikuchi diffraction is an emerging technique aimed at producing orientation maps of the structure of materials with a nanometric lateral resolution. This study investigates experimentally the depth resolution of the on-axis configuration, via a twinned silicon bi-crystal sample specifically designed and fabricated. The measured depth resolution varies from 30 to 65 nm in the range 10-30 keV, with a close to linear dependence with incident energy and no dependence with the total sample thickness. The depth resolution is explained in terms of two mechanisms acting concomitantly: generation of Kikuchi diffraction all along the thickness of the sample, associated with continuous absorption on the way out. A model based on the electron mean free path is used to account for the dependence with incident energy of the depth resolution. In addition, based on the results in silicon, the use of the mean absorption coefficient is proposed to predict the depth resolution for any atomic number and incident energy.

  12. The Growth of Protein Crystals Using McDUCK

    NASA Technical Reports Server (NTRS)

    Ewing, Felicia; Wilson, Lori; Nadarajah, Arunan; Pusey, Marc

    1998-01-01

    Most of the current microgravity crystal growth hardware is optimized to produce crystals within the limited time available on orbit. This often results in the actual nucleation and growth process being rushed or the system not coming to equilibrium within the limited time available. Longer duration hardware exists, but one cannot readily pick out crystals grown early versus those which nucleated and grew more slowly. We have devised a long duration apparatus, the Multi-chamber Dialysis Unit for Crystallization Kinetics, or McDUCK. This apparatus-is a series of protein chambers, stacked upon a precipitant reservoir chamber. All chambers are separated by a dialysis membrane, which serves to pass small molecules while retaining the protein. The volume of the Precipitant chamber is equal to the sum of the volumes of the protein chamber. In operation, the appropriate chambers are filled with precipitant solution or protein solution, and the McDUCK is placed standing upright, with the precipitant chamber on the bottom. The precipitant diffuses upwards over time, with the time to reach equilibration a function of the diffusivity of the precipitant and the overall length of the diffusion pathway. Typical equilibration times are approximately 2-4 months, and one can readily separate rapid from slow nucleation and growth crystals. An advantage on Earth is that the vertical precipitant concentration gradient dominates that of the solute, thus dampening out solute density gradient driven convective flows. However, large Earth-grown crystals have so far tended to be more two dimensional. Preliminary X-ray diffraction analysis of lysozyme crystals grown in McDUCK have indicated that the best, and largest, come from the middle chambers, suggesting that there is an optimal growth rate. Further, the improvements in diffraction resolution have been better signal to noise ratios in the low resolution data, not an increase in resolution overall. Due to the persistently large crystals grown we are currently proposing McDUCK for the growth of macromolecule crystals for use in neutron diffraction studies.

  13. [Detection of single-walled carbon nanotube bundles by tip-enhanced Raman spectroscopy].

    PubMed

    Wu, Xiao-Bin; Wang, Jia; Wang, Rui; Xu, Ji-Ying; Tian, Qian; Yu, Jian-Yuan

    2009-10-01

    Raman spectroscopy is a powerful technique in the characterization of carbon nanotubes (CNTs). However, this spectral method is subject to two obstacles. One is spatial resolution, namely the diffraction limits of light, and the other is its inherent small Raman cross section and weak signal. To resolve these problems, a new approach has been developed, denoted tip-enhanced Raman spectroscopy (TERS). TERS has been demonstrated to be a powerful spectroscopic and microscopic technique to characterize nanomaterial or nanostructures. Excited by a focused laser beam, an enhanced electric field is generated in the vicinity of a metallic tip because of the surface plasmon polariton (SPP) and lightening rod effect. Consequently, Raman signal from the sample area illuminated by the enhanced field nearby the tip is enhanced. At the same time, the topography is obtained in the nanometer scale. The exact corresponding relationship between the localized Raman and the topography makes the Raman identification at the nanometer scale to be feasible. In the present paper, based on an inverted microscope and a metallic AFM tip, a tip-enhanced Raman system was set up. The radius of the Au-coated metallic tip is about 30 nm. The 532 nm laser passes through a high numerical objective (NA0.95) from the bottom to illuminate the tip to excite the enhanced electric field. Corresponding with the AFM image, the tip-enhanced near-field Raman of a 100 nm diameter single-walled carbon nanotube (SWNT) bundles was obtained. The SWNTs were prepared by arc method. Furthermore, the near-field Raman of about 3 SWNTs of the bundles was received with the spatial resolution beyond the diffraction limit. Compared with the far-field Raman, the enhancement factor of the tip-enhanced Raman is more than 230. With the super-diffraction spatial resolution and the tip-enhanced Raman ability, tip-enhanced Raman spectroscopy will play an important role in the nano-material and nano-structure characterization.

  14. Characterization of Differential Toll-Like Receptor Responses below the Optical Diffraction Limit**

    PubMed Central

    Aaron, Jesse S.; Carson, Bryan D.; Timlin, Jerilyn A.

    2013-01-01

    Many membrane receptors are recruited to specific cell surface domains to form nanoscale clusters upon ligand activation. This step appears to be necessary to initiate signaling, including pathways in innate immune system activation. However, virulent pathogens such as Yersinia pestis (the causative agent of plague) are known to evade innate immune detection, in contrast to similar microbes (such as E. coli) that elicit a robust response. This disparity has been partly attributed to the structure of lipopolysaccharides (LPS) on the bacterial cell wall, which are recognized by the innate immune receptor TLR4. As such, we hypothesized that nanoscale differences would exist between the spatial clustering of TLR4 upon binding of LPS derived from Y. pestis and E. coli. Although optical imaging can provide exquisite details of the spatial organization of biomolecules, there is a mismatch between the scale at which receptor clustering occurs (<300 nm) and the optical diffraction limit (>400 nm). The last decade has seen the emergence of super-resolution imaging methods that effectively break the optical diffraction barrier to yield truly nanoscale information in intact biological samples. This study reports the first visualizations of TLR4 distributions on intact cells at image resolutions of <30 nm using a novel, dual-color stochastic optical reconstruction microscopy (STORM) technique. This methodology permits distinction between receptors containing bound LPS from those without at the nanoscale. Importantly, we also show that LPS derived from immuno-stimulatory bacteria resulted in significantly higher LPS-TLR4 cluster sizes and a nearly two-fold greater ligand/receptor colocalization as compared to immuno-evading LPS. PMID:22807232

  15. High resolution middle infrared spectrometer, a part of atmospheric chemistry suite (ACS) for EXOMARS 2016 trace gas orbiter

    NASA Astrophysics Data System (ADS)

    Trokhimovskiy, Alexander; Korablev, Oleg; Ivanov, Yurii; Syniavskyi, Ivan; Montmessin, Franck; Fedorova, Anna

    2017-11-01

    The Atmospheric Chemistry Suite (ACS) package is a part of Russian contribution to ExoMars ESARoscosmos mission for studies of the Martian atmosphere and climate. ACS consists of three separate infrared spectrometers, sharing common mechanical, electrical, and thermal interfaces. The mid-infrared (MIR) channel is a cross-dispersion high resolution echelle instrument dedicated to solar occultation measurements and sensitive studies of trace gases. The MIR channel is a spectrometer working in 2.3-4.2 μm spectral range, covering simultaneously up to almost 300 nm per exposure, targeting the resolving power of 50,000. A crossdispersion concept on echelle and ordinary diffraction grating allows acquisition of the wide wavelength domain at once. That provides a strategic advantage for maximizing the number of gaseous species detected simultaneously and good special resolution of measurements during fast occultation sessions. Moving the second grating allows to switch from one group of the diffraction orders to another prior to a series of measurements, or desired positions during one measurement sequence. The concept of the cross-dispersion echelle instrument, which is widely accepted in astronomy, has been already employed in planetary missions with VIRTIS-H instrument presently in flight on Rosetta and Venus Express missions. Targeting very high spectral resolution the MIR channel operates in solar occultation only. A telescope with relative aperture of 1∶3 forms the image of the solar disk on the slit. The FOV is determined by the slit and it consists 0.1×2.9 mrad. The spectral resolution of the spectrometer is fully slit-limited, and the resolving power of λ/Δλ >= 50000 at 3.3 μm is supported. Two secondary cross-dispersion diffraction gratings (plain, 180 and 361 grooves per mm) are mounted back-to-back on a stepper motor to change observed echelle orders. We have chosen two secondary gratings philosophy to switch between them depending on the long or short wavelength range we are on. Changing the position of the secondary grating in angular steps of 1.8°, from 10 to 30 echelle orders are available for simultaneous record depending on the wavelength. 100 steps are evidently used to switch between gratings prior measurements. The full spectral range is covered on diffraction orders from 142 to 248. For each observation detector area is covered by 10 to 30 stripes, each corresponding to single echelle diffraction order. Given the complexity of the diffraction orders pattern, full detector frames will be transmitted to the ground, with lossless compression. However, the onboard averaging will be possible. Single data frame will be accumulated for 0.5 seconds, stacking of a number of shorter exposures.

  16. Depth-resolved mid-infrared photothermal imaging of living cells and organisms with submicrometer spatial resolution

    PubMed Central

    Zhang, Delong; Li, Chen; Zhang, Chi; Slipchenko, Mikhail N.; Eakins, Gregory; Cheng, Ji-Xin

    2016-01-01

    Chemical contrast has long been sought for label-free visualization of biomolecules and materials in complex living systems. Although infrared spectroscopic imaging has come a long way in this direction, it is thus far only applicable to dried tissues because of the strong infrared absorption by water. It also suffers from low spatial resolution due to long wavelengths and lacks optical sectioning capabilities. We overcome these limitations through sensing vibrational absorption–induced photothermal effect by a visible laser beam. Our mid-infrared photothermal (MIP) approach reached 10 μM detection sensitivity and submicrometer lateral spatial resolution. This performance has exceeded the diffraction limit of infrared microscopy and allowed label-free three-dimensional chemical imaging of live cells and organisms. Distributions of endogenous lipid and exogenous drug inside single cells were visualized. We further demonstrated in vivo MIP imaging of lipids and proteins in Caenorhabditis elegans. The reported MIP imaging technology promises broad applications from monitoring metabolic activities to high-resolution mapping of drug molecules in living systems, which are beyond the reach of current infrared microscopy. PMID:27704043

  17. Depth-resolved mid-infrared photothermal imaging of living cells and organisms with submicrometer spatial resolution.

    PubMed

    Zhang, Delong; Li, Chen; Zhang, Chi; Slipchenko, Mikhail N; Eakins, Gregory; Cheng, Ji-Xin

    2016-09-01

    Chemical contrast has long been sought for label-free visualization of biomolecules and materials in complex living systems. Although infrared spectroscopic imaging has come a long way in this direction, it is thus far only applicable to dried tissues because of the strong infrared absorption by water. It also suffers from low spatial resolution due to long wavelengths and lacks optical sectioning capabilities. We overcome these limitations through sensing vibrational absorption-induced photothermal effect by a visible laser beam. Our mid-infrared photothermal (MIP) approach reached 10 μM detection sensitivity and submicrometer lateral spatial resolution. This performance has exceeded the diffraction limit of infrared microscopy and allowed label-free three-dimensional chemical imaging of live cells and organisms. Distributions of endogenous lipid and exogenous drug inside single cells were visualized. We further demonstrated in vivo MIP imaging of lipids and proteins in Caenorhabditis elegans . The reported MIP imaging technology promises broad applications from monitoring metabolic activities to high-resolution mapping of drug molecules in living systems, which are beyond the reach of current infrared microscopy.

  18. Pushing the limits of sulfur SAD phasing: de novo structure solution of the N-terminal domain of the ectodomain of HCV E1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    El Omari, Kamel; Iourin, Oleg; Kadlec, Jan

    2014-08-01

    The sulfur SAD phasing method was successfully used to determine the structure of the N-terminal domain of HCV E1 from low-resolution diffracting crystals by combining data from 32 crystals. Single-wavelength anomalous dispersion of S atoms (S-SAD) is an elegant phasing method to determine crystal structures that does not require heavy-atom incorporation or selenomethionine derivatization. Nevertheless, this technique has been limited by the paucity of the signal at the usual X-ray wavelengths, requiring very accurate measurement of the anomalous differences. Here, the data collection and structure solution of the N-terminal domain of the ectodomain of HCV E1 from crystals that diffractedmore » very weakly is reported. By combining the data from 32 crystals, it was possible to solve the sulfur substructure and calculate initial maps at 7 Å resolution, and after density modication and phase extension using a higher resolution native data set to 3.5 Å resolution model building was achievable.« less

  19. Squeezing on Momentum States for Atom Interferometry.

    PubMed

    Salvi, Leonardo; Poli, Nicola; Vuletić, Vladan; Tino, Guglielmo M

    2018-01-19

    We propose and analyze a method that allows for the production of squeezed states of the atomic center-of-mass motion that can be injected into an atom interferometer. Our scheme employs dispersive probing in a ring resonator on a narrow transition in order to provide a collective measurement of the relative population of two momentum states. We show that this method is applicable to a Bragg diffraction-based strontium atom interferometer with large diffraction orders. This technique can be extended also to small diffraction orders and large atom numbers N by inducing atomic transparency at the frequency of the probe field, reaching an interferometer phase resolution scaling Δϕ∼N^{-3/4}. We show that for realistic parameters it is possible to obtain a 20 dB gain in interferometer phase estimation compared to the standard quantum limit. Our method is applicable to other atomic species where a narrow transition is available or can be synthesized.

  20. Pink-beam serial crystallography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Meents, A.; Wiedorn, M. O.; Srajer, V.

    Serial X-ray crystallography allows macromolecular structure determination at both X-ray free electron lasers (XFELs) and, more recently, synchrotron sources. The time resolution for serial synchrotron crystallography experiments has been limited to millisecond timescales with monochromatic beams. The polychromatic, “pink”, beam provides a more than two orders of magnitude increased photon flux and hence allows accessing much shorter timescales in diffraction experiments at synchrotron sources. Here we report the structure determination of two different protein samples by merging pink-beam diffraction patterns from many crystals, each collected with a single 100 ps X-ray pulse exposure per crystal using a setup optimized formore » very low scattering background. In contrast to experiments with monochromatic radiation, data from only 50 crystals were required to obtain complete datasets. The high quality of the diffraction data highlights the potential of this method for studying irreversible reactions at sub-microsecond timescales using high-brightness X-ray facilities.« less

  1. Pink-beam serial crystallography

    DOE PAGES

    Meents, A.; Wiedorn, M. O.; Srajer, V.; ...

    2017-11-03

    Serial X-ray crystallography allows macromolecular structure determination at both X-ray free electron lasers (XFELs) and, more recently, synchrotron sources. The time resolution for serial synchrotron crystallography experiments has been limited to millisecond timescales with monochromatic beams. The polychromatic, “pink”, beam provides a more than two orders of magnitude increased photon flux and hence allows accessing much shorter timescales in diffraction experiments at synchrotron sources. Here we report the structure determination of two different protein samples by merging pink-beam diffraction patterns from many crystals, each collected with a single 100 ps X-ray pulse exposure per crystal using a setup optimized formore » very low scattering background. In contrast to experiments with monochromatic radiation, data from only 50 crystals were required to obtain complete datasets. The high quality of the diffraction data highlights the potential of this method for studying irreversible reactions at sub-microsecond timescales using high-brightness X-ray facilities.« less

  2. RESOLFT nanoscopy with photoswitchable organic fluorophores

    NASA Astrophysics Data System (ADS)

    Kwon, Jiwoong; Hwang, Jihee; Park, Jaewan; Han, Gi Rim; Han, Kyu Young; Kim, Seong Keun

    2015-12-01

    Far-field optical nanoscopy has been widely used to image small objects with sub-diffraction-limit spatial resolution. Particularly, reversible saturable optical fluorescence transition (RESOLFT) nanoscopy with photoswitchable fluorescent proteins is a powerful method for super-resolution imaging of living cells with low light intensity. Here we demonstrate for the first time the implementation of RESOLFT nanoscopy for a biological system using organic fluorophores, which are smaller in size and easier to be chemically modified. With a covalently-linked dye pair of Cy3 and Alexa647 to label subcellular structures in fixed cells and by optimizing the imaging buffer and optical parameters, our RESOLFT nanoscopy achieved a spatial resolution of ~74 nm in the focal plane. This method provides a powerful alternative for low light intensity RESOLFT nanoscopy, which enables biological imaging with small organic probes at nanoscale resolution.

  3. Terahertz-wave near-field imaging with subwavelength resolution using surface-wave-assisted bow-tie aperture

    NASA Astrophysics Data System (ADS)

    Ishihara, Kunihiko; Ohashi, Keishi; Ikari, Tomofumi; Minamide, Hiroaki; Yokoyama, Hiroyuki; Shikata, Jun-ichi; Ito, Hiromasa

    2006-11-01

    We demonstrate the terahertz-wave near-field imaging with subwavelength resolution using a bow-tie shaped aperture surrounded by concentric periodic structures in a metal film. A subwavelength aperture with concentric periodic grooves, which are known as a bull's eye structure, shows extremely large enhanced transmission beyond the diffraction limit caused by the resonant excitation of surface waves. Additionally, a bow-tie aperture exhibits extraordinary field enhancement at the sharp tips of the metal, which enhances the transmission and the subwavelength spatial resolution. We introduced a bow-tie aperture to the bull's eye structure and achieved high spatial resolution (˜λ/17) in the near-field region. The terahertz-wave near-field image of the subwavelength metal pattern (pattern width=20μm) was obtained for the wavelength of 207μm.

  4. Micrometer-resolution imaging using MÖNCH: towards G2-less grating interferometry

    PubMed Central

    Cartier, Sebastian; Kagias, Matias; Bergamaschi, Anna; Wang, Zhentian; Dinapoli, Roberto; Mozzanica, Aldo; Ramilli, Marco; Schmitt, Bernd; Brückner, Martin; Fröjdh, Erik; Greiffenberg, Dominic; Mayilyan, Davit; Mezza, Davide; Redford, Sophie; Ruder, Christian; Schädler, Lukas; Shi, Xintian; Thattil, Dhanya; Tinti, Gemma; Zhang, Jiaguo; Stampanoni, Marco

    2016-01-01

    MÖNCH is a 25 µm-pitch charge-integrating detector aimed at exploring the limits of current hybrid silicon detector technology. The small pixel size makes it ideal for high-resolution imaging. With an electronic noise of about 110 eV r.m.s., it opens new perspectives for many synchrotron applications where currently the detector is the limiting factor, e.g. inelastic X-ray scattering, Laue diffraction and soft X-ray or high-resolution color imaging. Due to the small pixel pitch, the charge cloud generated by absorbed X-rays is shared between neighboring pixels for most of the photons. Therefore, at low photon fluxes, interpolation algorithms can be applied to determine the absorption position of each photon with a resolution of the order of 1 µm. In this work, the characterization results of one of the MÖNCH prototypes are presented under low-flux conditions. A custom interpolation algorithm is described and applied to the data to obtain high-resolution images. Images obtained in grating interferometry experiments without the use of the absorption grating G2 are shown and discussed. Perspectives for the future developments of the MÖNCH detector are also presented. PMID:27787252

  5. Quantitative evaluation of high-resolution features in images of negatively stained Tobacco Mosaic Virus.

    PubMed

    Chang, C F; Williams, R C; Grano, D A; Downing, K H; Glaeser, R M

    1983-01-01

    This study investigates the causes of the apparent differences between the optical diffraction pattern of a micrograph of a Tobacco Mosaic Virus (TMV) particle, the optical diffraction pattern of a ten-fold photographically averaged image, and the computed diffraction pattern of the original micrograph. Peak intensities along the layer lines in the transform of the averaged image appear to be quite unlike those in the diffraction pattern of the original micrograph, and the diffraction intensities for the averaged image extend to unexpectedly high resolution. A carefully controlled, quantitative comparison reveals, however, that the optical diffraction pattern of the original micrograph and that of the ten-fold averaged image are essentially equivalent. Using computer-based image processing, we discovered that the peak intensities on the 6th layer line have values very similar in magnitude to the neighboring noise, in contrast to what was expected from the optical diffraction pattern of the original micrograph. This discrepancy was resolved by recording a series of optical diffraction patterns when the original micrograph was immersed in oil. These patterns revealed the presence of a substantial phase grating effect, which exaggerated the peak intensities on the 6th layer line, causing an erroneous impression that the high resolution features possessed a good signal-to-noise ratio. This study thus reveals some pitfalls and misleading results that can be encountered when using optical diffraction patterns to evaluate image quality.

  6. Objective Lens Optimized for Wavefront Delivery, Pupil Imaging, and Pupil Ghosting

    NASA Technical Reports Server (NTRS)

    Olzcak, Gene

    2009-01-01

    An interferometer objective lens (or diverger) may be used to transform a collimated beam into a diverging or converging beam. This innovation provides an objective lens that has diffraction-limited optical performance that is optimized at two sets of conjugates: imaging to the objective focus and imaging to the pupil. The lens thus provides for simultaneous delivery of a high-quality beam and excellent pupil resolution properties.

  7. Target Acquisition for Projectile Vision-Based Navigation

    DTIC Science & Technology

    2014-03-01

    Future Work 20 8. References 21 Appendix A. Simulation Results 23 Appendix B. Derivation of Ground Resolution for a Diffraction-Limited Pinhole Camera...results for visual acquisition (left) and target recognition (right). ..........19 Figure B-1. Differential object and image areas for pinhole camera...projectile and target (measured in terms of the angle ) will depend on target heading. In particular, because we have aligned the x axis along the

  8. Micromachined optical microphone structures with low thermal-mechanical noise levels.

    PubMed

    Hall, Neal A; Okandan, Murat; Littrell, Robert; Bicen, Baris; Degertekin, F Levent

    2007-10-01

    Micromachined microphones with diffraction-based optical displacement detection have been introduced previously [Hall et al., J. Acoust. Soc. Am. 118, 3000-3009 (2005)]. The approach has the advantage of providing high displacement detection resolution of the microphone diaphragm independent of device size and capacitance-creating an unconstrained design space for the mechanical structure itself. Micromachined microphone structures with 1.5-mm-diam polysilicon diaphragms and monolithically integrated diffraction grating electrodes are presented in this work with backplate architectures that deviate substantially from traditional perforated plate designs. These structures have been designed for broadband frequency response and low thermal mechanical noise levels. Rigorous experimental characterization indicates a diaphragm displacement detection resolution of 20 fm radicalHz and a thermal mechanical induced diaphragm displacement noise density of 60 fm radicalHz, corresponding to an A-weighted sound pressure level detection limit of 24 dB(A) for these structures. Measured thermal mechanical displacement noise spectra are in excellent agreement with simulations based on system parameters derived from dynamic frequency response characterization measurements, which show a diaphragm resonance limited bandwidth of approximately 20 kHz. These designs are substantial improvements over initial prototypes presented previously. The high performance-to-size ratio achievable with this technology is expected to have an impact on a variety of instrumentation and hearing applications.

  9. Thaumatin crystallization aboard the International Space Station using liquid-liquid diffusion in the Enhanced Gaseous Nitrogen Dewar (EGN).

    PubMed

    Barnes, Cindy L; Snell, Edward H; Kundrot, Craig E

    2002-05-01

    This paper reports results from the first biological crystal-growth experiment on the International Space Station (ISS). Crystals of thaumatin were grown using liquid-liquid diffusion in Tygon tubing transported in the Enhanced Gaseous Nitrogen Dewar (EGN). Different volume ratios and concentrations of protein and precipitant were used to test different adaptations of the vapor-diffusion crystallization recipe to the liquid-liquid diffusion method. The EGN warmed up from 77 to 273 K in about 4 d, about the same time it took to warm from 273 to 293 K. The temperature within the EGN was 293-297 K for the majority of the experiment. Air gaps that blocked liquid-liquid diffusion formed in the tubes. Nonetheless, crystals were grown. Synchrotron diffraction data collected from the best space-grown crystal extended to 1.28 A, comparable to previous studies of space-grown thaumatin crystals. The resolution of the best ground-control crystal was only 1.47 A. It is not clear if the difference in diffraction limit arises from factors other than crystal size. Improvements in temperature control and the elimination of air gaps are needed, but the results show that the EGN on the ISS can be used to produce space-grown crystals that diffract to high resolution.

  10. Thaumatin Crystallization Aboard the International Space Station Using Liquid-Liquid Diffusion in the Enhanced Gaseous Nitrogen Dewar (EGN)

    NASA Technical Reports Server (NTRS)

    Kundrot, Craig; Barnes, Cindy L.; Snell, Edward H.; Stinson, Thomas N. (Technical Monitor)

    2002-01-01

    This paper reports results from the first biological crystal growth experiment on the International Space Station (ISS). Crystals of thaumatin were grown using liquid-liquid diffusion in Tygon tubing transported in the Enhanced Gaseous Nitrogen Dewar (EGN). Different Volume ratios and concentrations of protein and precipitant were used to test different adaptations of the vapor diffusion crystallization recipe to the liquid-liquid diffusion method. The EGN warmed up from -196 C to 0 C in about four days, about the same time it took to warm from 0 C to 20 C. The temperature within the EGN was 20 - 24 C for the majority of the experiment. Air gaps that blocked liquid-liquid diffusion formed in the tubes. Nonetheless, crystals were grown. Synchrotron diffraction data collected from the best space grown crystal extended to 1.28 Angstroms, comparable to previous studies of space-grown thaumatin crystals. The resolution of the best ground control crystal was only 1.47 Angstroms. It is not clear if the difference in diffraction limit is due to factors other than crystal size. Improvements in temperature control and the elimination of air gaps are needed, but the results show that EGN on the ISS can be used to produce space grown crystals that diffract to high resolution.

  11. Crystallization of the C-terminal domain of the addiction antidote CcdA in complex with its toxin CcdB

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Buts, Lieven; De Jonge, Natalie; Loris, Remy, E-mail: reloris@vub.ac.be

    2005-10-01

    The CcdA C-terminal domain was crystallized in complex with CcdB in two crystal forms that diffract to beyond 2.0 Å resolution. CcdA and CcdB are the antidote and toxin of the ccd addiction module of Escherichia coli plasmid F. The CcdA C-terminal domain (CcdA{sub C36}; 36 amino acids) was crystallized in complex with CcdB (dimer of 2 × 101 amino acids) in three different crystal forms, two of which diffract to high resolution. Form II belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 37.6, b = 60.5, c = 83.8 Å and diffracts to 1.8more » Å resolution. Form III belongs to space group P2{sub 1}, with unit-cell parameters a = 41.0, b = 37.9, c = 69.6 Å, β = 96.9°, and diffracts to 1.9 Å resolution.« less

  12. Direct Detection Electron Energy-Loss Spectroscopy: A Method to Push the Limits of Resolution and Sensitivity.

    PubMed

    Hart, James L; Lang, Andrew C; Leff, Asher C; Longo, Paolo; Trevor, Colin; Twesten, Ray D; Taheri, Mitra L

    2017-08-15

    In many cases, electron counting with direct detection sensors offers improved resolution, lower noise, and higher pixel density compared to conventional, indirect detection sensors for electron microscopy applications. Direct detection technology has previously been utilized, with great success, for imaging and diffraction, but potential advantages for spectroscopy remain unexplored. Here we compare the performance of a direct detection sensor operated in counting mode and an indirect detection sensor (scintillator/fiber-optic/CCD) for electron energy-loss spectroscopy. Clear improvements in measured detective quantum efficiency and combined energy resolution/energy field-of-view are offered by counting mode direct detection, showing promise for efficient spectrum imaging, low-dose mapping of beam-sensitive specimens, trace element analysis, and time-resolved spectroscopy. Despite the limited counting rate imposed by the readout electronics, we show that both core-loss and low-loss spectral acquisition are practical. These developments will benefit biologists, chemists, physicists, and materials scientists alike.

  13. Resolution of 90 nm (lambda/5) in an optical transmission microscope with an annular condenser.

    PubMed

    Vainrub, Arnold; Pustovyy, Oleg; Vodyanoy, Vitaly

    2006-10-01

    Resolution of 90 nm was achieved with a research microscope simply by replacing the standard bright-field condenser with a homebuilt illumination system with a cardioid annular condenser. Diffraction gratings with 100 nm width lines as well as less than 100 nm size features of different-shaped objects were clearly visible on a calibrated microscope test slide. The resolution increase results from a known narrower diffraction pattern in coherent illumination for the annular aperture compared with the circular aperture. This explanation is supported by an excellent accord of calculated and measured diffraction patterns for a 50 nm radius disk.

  14. Diffraction encoded position measuring apparatus

    DOEpatents

    Tansey, Richard J.

    1991-01-01

    When a lightwave passes through a transmission grating, diffracted beams appear at the output or opposite side of the grating that are effectively Doppler shifted in frequency (phase) whereby a detector system can compare the phase of the zero order and higher order beams to obtain an indication of position. Multiple passes through the grating increase resolution for a given wavelength of a laser signal. The resolution can be improved further by using a smaller wavelength laser to generate the grating itself. Since the grating must only have a pitch sufficient to produce diffracted orders, inexpensive, ultraviolet wavelength lasers can be utilized and still obtain high resolution detection.

  15. Diffraction encoded position measuring apparatus

    DOEpatents

    Tansey, R.J.

    1991-09-24

    When a lightwave passes through a transmission grating, diffracted beams appear at the output or opposite side of the grating that are effectively Doppler shifted in frequency (phase) whereby a detector system can compare the phase of the zero order and higher order beams to obtain an indication of position. Multiple passes through the grating increase resolution for a given wavelength of a laser signal. The resolution can be improved further by using a smaller wavelength laser to generate the grating itself. Since the grating must only have a pitch sufficient to produce diffracted orders, inexpensive, ultraviolet wavelength lasers can be utilized and still obtain high resolution detection. 3 figures.

  16. Crystallization of multiple forms of bovine seminal ribonuclease in the liganded and unliganded state

    NASA Astrophysics Data System (ADS)

    Sica, F.; Adinolfi, S.; Berisio, R.; De Lorenzo, C.; Mazzarella, L.; Piccoli, R.; Vitagliano, L.; Zagari, A.

    1999-01-01

    Bovine seminal ribonuclease (BS-RNase) is an intriguing homodimeric enzyme which exists as two conformational isomers, characterized by distinct catalytic and biological properties, referred to as M×M and M=M. Reduction of inter-chain disulfide bridges produces a stable monomeric derivative (M) which is still active. This paper reports the screening and optimization of crystallization conditions for growing single diffraction-quality crystals for the various BS-RNase forms. The crystallization trials were performed using both the vapor diffusion and microbatch methods. The M×M dimer was crystallized in the free form from polyethylene glycol (PEG) 4000 at pH 8.5 and as a complex with the substrate analog uridylyl(2'- 5')guanosine (UpG) from an unbuffered ammonium sulfate (AS) solution. These two crystal types diffract X-rays to 2.5 and 1.9 Å resolution, respectively. Two different crystal types were obtained both for the M=M dimer and for the monomeric derivative. (M=M)a crystals, grown from PEG 4000 (8% w/v) at pH 5.6, diffract X-rays to 4.0 Å. At higher PEG concentration (15% w/v) a different crystal type was obtained, (M=M)b, which showed a better diffraction limit (2.5 Å). For the monomer, type (M)a and (M)b crystals, diffracting X-rays to 2.5 Å resolution, were obtained from AS at pH 6.5 and from PEG 4000 at pH 8.5, respectively. A comparison with previously crystallized forms of the dimer M×M and its complexes with uridylyl(2'-5')adenosine and 2'-deoxycytidylyl(3'-5')-2'-deoxyadenosine is also presented. The three-dimensional structure analysis of (M×M)·UpG and (M=M)b is in progress.

  17. Imaging live cells at high spatiotemporal resolution for lab-on-a-chip applications.

    PubMed

    Chin, Lip Ket; Lee, Chau-Hwang; Chen, Bi-Chang

    2016-05-24

    Conventional optical imaging techniques are limited by the diffraction limit and difficult-to-image biomolecular and sub-cellular processes in living specimens. Novel optical imaging techniques are constantly evolving with the desire to innovate an imaging tool that is capable of seeing sub-cellular processes in a biological system, especially in three dimensions (3D) over time, i.e. 4D imaging. For fluorescence imaging on live cells, the trade-offs among imaging depth, spatial resolution, temporal resolution and photo-damage are constrained based on the limited photons of the emitters. The fundamental solution to solve this dilemma is to enlarge the photon bank such as the development of photostable and bright fluorophores, leading to the innovation in optical imaging techniques such as super-resolution microscopy and light sheet microscopy. With the synergy of microfluidic technology that is capable of manipulating biological cells and controlling their microenvironments to mimic in vivo physiological environments, studies of sub-cellular processes in various biological systems can be simplified and investigated systematically. In this review, we provide an overview of current state-of-the-art super-resolution and 3D live cell imaging techniques and their lab-on-a-chip applications, and finally discuss future research trends in new and breakthrough research areas of live specimen 4D imaging in controlled 3D microenvironments.

  18. Final Technical Report for DE-SC0008149

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Buchanan, Kristen

    The major goal of this project is to study spin waves in magnetic thin films, especially how spin waves respond to external stimuli. This is expected to lead to new insight into dynamic processes and new ideas for methods to control spin waves. Experimental studies are being done primarily using time- and spatially-resolved Brillouin light scattering (BLS) measurements on extended and patterned magnetic thin films. BLS is a versatile tool that provides a non-invasive probe of spin dynamics with frequencies of ~1 GHz to well over 100 GHz, diffraction-limited spatial resolution, 250-ps temporal resolution, and it is sensitive enough tomore » detect thermal magnons.« less

  19. Flatland Photonics: Circumventing Diffraction with Planar Plasmonic Architectures

    NASA Astrophysics Data System (ADS)

    Dionne, Jennifer Anne

    On subwavelength scales, photon-matter interactions are limited by diffraction. The diffraction limit restricts the size of optical devices and the resolution of conventional microscopes to wavelength-scale dimensions, severely hampering our ability to control and probe subwavelength-scale optical phenomena. Circumventing diffraction is now a principle focus of integrated nanophotonics. Surface plasmons provide a particularly promising approach to sub-diffraction-limited photonics. Surface plasmons are hybrid electron-photon modes confined to the interface between conductors and transparent materials. Combining the high localization of electronic waves with the propagation properties of optical waves, plasmons can achieve extremely small mode wavelengths and large local electromagnetic field intensities. Through their unique dispersion, surface plasmons provide access to an enormous phase space of refractive indices and propagation constants that can be readily tuned with material or geometry. In this thesis, we explore both the theory and applications of dispersion in planar plasmonic architectures. Particular attention is given to the modes of metallic core and plasmon slot waveguides, which can span positive, near-zero, and even negative indices. We demonstrate how such basic plasmonic geometries can be used to develop a suite of passive and active plasmonic components, including subwavelength waveguides, color filters, negative index metamaterials, and optical MOS field effect modulators. Positive index modes are probed by near- and far-field techniques, revealing plasmon wavelengths as small as one-tenth of the excitation wavelength. Negative index modes are characterized through direct visualization of negative refraction. By fabricating prisms comprised of gold, silicon nitride, and silver multilayers, we achieve the first experimental demonstration of a negative index material at visible frequencies, with potential applications for sub-diffraction-limited microscopy and electromagnetic cloaking. We exploit this tunability of complex plasmon mode indices to create a compact metal-oxide-Si (MOS) field effect plasmonic modulator (or plasMOStor). By transforming the MOS gate oxide into an optical channel, amplitude modulation depths of 11.2 dB are achieved in device volumes as small as one one-fifth of a cubic wavelength. Our results indicate the accessibility of tunable refractive indices over a wide frequency band, facilitating design of a new materials class with extraordinary optical properties and applications.

  20. Exploring the atomic structure and conformational flexibility of a 320 Å long engineered viral fiber using X-ray crystallography.

    PubMed

    Bhardwaj, Anshul; Casjens, Sherwood R; Cingolani, Gino

    2014-02-01

    Protein fibers are widespread in nature, but only a limited number of high-resolution structures have been determined experimentally. Unlike globular proteins, fibers are usually recalcitrant to form three-dimensional crystals, preventing single-crystal X-ray diffraction analysis. In the absence of three-dimensional crystals, X-ray fiber diffraction is a powerful tool to determine the internal symmetry of a fiber, but it rarely yields atomic resolution structural information on complex protein fibers. An 85-residue-long minimal coiled-coil repeat unit (MiCRU) was previously identified in the trimeric helical core of tail needle gp26, a fibrous protein emanating from the tail apparatus of the bacteriophage P22 virion. Here, evidence is provided that an MiCRU can be inserted in frame inside the gp26 helical core to generate a rationally extended fiber (gp26-2M) which, like gp26, retains a trimeric quaternary structure in solution. The 2.7 Å resolution crystal structure of this engineered fiber, which measures ∼320 Å in length and is only 20-35 Å wide, was determined. This structure, the longest for a trimeric protein fiber to be determined to such a high resolution, reveals the architecture of 22 consecutive trimerization heptads and provides a framework to decipher the structural determinants for protein fiber assembly, stability and flexibility.

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

    NASA Astrophysics Data System (ADS)

    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.

  2. Sub-diffraction Limit Localization of Proteins in Volumetric Space Using Bayesian Restoration of Fluorescence Images from Ultrathin Specimens

    PubMed Central

    Wang, Gordon; Smith, Stephen J.

    2012-01-01

    Photon diffraction limits the resolution of conventional light microscopy at the lateral focal plane to 0.61λ/NA (λ = wavelength of light, NA = numerical aperture of the objective) and at the axial plane to 1.4nλ/NA2 (n = refractive index of the imaging medium, 1.51 for oil immersion), which with visible wavelengths and a 1.4NA oil immersion objective is ∼220 nm and ∼600 nm in the lateral plane and axial plane respectively. This volumetric resolution is too large for the proper localization of protein clustering in subcellular structures. Here we combine the newly developed proteomic imaging technique, Array Tomography (AT), with its native 50–100 nm axial resolution achieved by physical sectioning of resin embedded tissue, and a 2D maximum likelihood deconvolution method, based on Bayes' rule, which significantly improves the resolution of protein puncta in the lateral plane to allow accurate and fast computational segmentation and analysis of labeled proteins. The physical sectioning of AT allows tissue specimens to be imaged at the physical optimum of modern high NA plan-apochormatic objectives. This translates to images that have little out of focus light, minimal aberrations and wave-front distortions. Thus, AT is able to provide images with truly invariant point spread functions (PSF), a property critical for accurate deconvolution. We show that AT with deconvolution increases the volumetric analytical fidelity of protein localization by significantly improving the modulation of high spatial frequencies up to and potentially beyond the spatial frequency cut-off of the objective. Moreover, we are able to achieve this improvement with no noticeable introduction of noise or artifacts and arrive at object segmentation and localization accuracies on par with image volumes captured using commercial implementations of super-resolution microscopes. PMID:22956902

  3. Sub-diffraction limit localization of proteins in volumetric space using Bayesian restoration of fluorescence images from ultrathin specimens.

    PubMed

    Wang, Gordon; Smith, Stephen J

    2012-01-01

    Photon diffraction limits the resolution of conventional light microscopy at the lateral focal plane to 0.61λ/NA (λ = wavelength of light, NA = numerical aperture of the objective) and at the axial plane to 1.4nλ/NA(2) (n = refractive index of the imaging medium, 1.51 for oil immersion), which with visible wavelengths and a 1.4NA oil immersion objective is -220 nm and -600 nm in the lateral plane and axial plane respectively. This volumetric resolution is too large for the proper localization of protein clustering in subcellular structures. Here we combine the newly developed proteomic imaging technique, Array Tomography (AT), with its native 50-100 nm axial resolution achieved by physical sectioning of resin embedded tissue, and a 2D maximum likelihood deconvolution method, based on Bayes' rule, which significantly improves the resolution of protein puncta in the lateral plane to allow accurate and fast computational segmentation and analysis of labeled proteins. The physical sectioning of AT allows tissue specimens to be imaged at the physical optimum of modern high NA plan-apochormatic objectives. This translates to images that have little out of focus light, minimal aberrations and wave-front distortions. Thus, AT is able to provide images with truly invariant point spread functions (PSF), a property critical for accurate deconvolution. We show that AT with deconvolution increases the volumetric analytical fidelity of protein localization by significantly improving the modulation of high spatial frequencies up to and potentially beyond the spatial frequency cut-off of the objective. Moreover, we are able to achieve this improvement with no noticeable introduction of noise or artifacts and arrive at object segmentation and localization accuracies on par with image volumes captured using commercial implementations of super-resolution microscopes.

  4. Producing superfluid circulation states using phase imprinting

    NASA Astrophysics Data System (ADS)

    Kumar, Avinash; Dubessy, Romain; Badr, Thomas; De Rossi, Camilla; de Goër de Herve, Mathieu; Longchambon, Laurent; Perrin, Hélène

    2018-04-01

    We propose a method to prepare states of given quantized circulation in annular Bose-Einstein condensates (BEC) confined in a ring trap using the method of phase imprinting without relying on a two-photon angular momentum transfer. The desired phase profile is imprinted on the atomic wave function using a short light pulse with a tailored intensity pattern generated with a spatial light modulator. We demonstrate the realization of "helicoidal" intensity profiles suitable for this purpose. Due to the diffraction limit, the theoretical steplike intensity profile is not achievable in practice. We investigate the effect of imprinting an intensity profile smoothed by a finite optical resolution onto the annular BEC with a numerical simulation of the time-dependent Gross-Pitaevskii equation. This allows us to optimize the intensity pattern for a given target circulation to compensate for the limited resolution.

  5. HYBRID simulations of diffraction-limited focusing with Kirkpatrick-Baez mirrors for a next-generation In Situ hard X-ray nanoprobe

    DOE PAGES

    Maser, Jorg; Shi, Xianbo; Reininger, Ruben; ...

    2016-02-22

    Next-generation hard X-ray nanoprobe beamlines such as the In Situ Nanoprobe (ISN) beamline being planned at the Advanced Photon Source aim at providing very high spatial resolution while also enabling very high focused flux, to study complex materials and devices using fast, multidimensional imaging across many length scales. The ISN will use diffractive optics to focus X-rays with a bandpass of ΔE/E = 10 –4 into a focal spot of 20 nm or below. Reflective optics in Kirkpatrick-Baez geometry will be used to focus X-rays with a bandpass as large as ΔE/E = 10 –2 into a focal spot ofmore » 50 nm. Diffraction-limited focusing with reflective optics is achieved by spatial filtering and use of a very long, vertically focusing mirror. Furthermore, to quantify the performance of the ISN beamline, we have simulated the propagation of both partially and fully coherent wavefronts from the undulator source, through the ISN beamline and into the mirror-based focal spot. Simulations were carried out using the recently developed software “HYBRID.”« less

  6. Combined operando X-ray diffraction-electrochemical impedance spectroscopy detecting solid solution reactions of LiFePO4 in batteries.

    PubMed

    Hess, Michael; Sasaki, Tsuyoshi; Villevieille, Claire; Novák, Petr

    2015-09-08

    Lithium-ion batteries are widely used for portable applications today; however, often suffer from limited recharge rates. One reason for such limitation can be a reduced active surface area during phase separation. Here we report a technique combining high-resolution operando synchrotron X-ray diffraction coupled with electrochemical impedance spectroscopy to directly track non-equilibrium intermediate phases in lithium-ion battery materials. LiFePO4, for example, is known to undergo phase separation when cycled under low-current-density conditions. However, operando X-ray diffraction under ultra-high-rate alternating current and direct current excitation reveal a continuous but current-dependent, solid solution reaction between LiFePO4 and FePO4 which is consistent with previous experiments and calculations. In addition, the formation of a preferred phase with a composition similar to the eutectoid composition, Li0.625FePO4, is evident. Even at a low rate of 0.1C, ∼20% of the X-ray diffractogram can be attributed to non-equilibrium phases, which changes our understanding of the intercalation dynamics in LiFePO4.

  7. Dark-field imaging based on post-processed electron backscatter diffraction patterns of bulk crystalline materials in a scanning electron microscope.

    PubMed

    Brodusch, Nicolas; Demers, Hendrix; Gauvin, Raynald

    2015-01-01

    Dark-field (DF) images were acquired in the scanning electron microscope with an offline procedure based on electron backscatter diffraction (EBSD) patterns (EBSPs). These EBSD-DF images were generated by selecting a particular reflection on the electron backscatter diffraction pattern and by reporting the intensity of one or several pixels around this point at each pixel of the EBSD-DF image. Unlike previous studies, the diffraction information of the sample is the basis of the final image contrast with a pixel scale resolution at the EBSP providing DF imaging in the scanning electron microscope. The offline facility of this technique permits the selection of any diffraction condition available in the diffraction pattern and displaying the corresponding image. The high number of diffraction-based images available allows a better monitoring of deformation structures compared to electron channeling contrast imaging (ECCI) which is generally limited to a few images of the same area. This technique was applied to steel and iron specimens and showed its high capability in describing more rigorously the deformation structures around micro-hardness indents. Due to the offline relation between the reference EBSP and the EBSD-DF images, this new technique will undoubtedly greatly improve our knowledge of deformation mechanism and help to improve our understanding of the ECCI contrast mechanisms. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. The Influence of Surface Morphology and Diffraction Resolution of Canavalin Crystals

    NASA Technical Reports Server (NTRS)

    Plomp, M.; Thomas, B. R.; Day, J. S.; McPherson, A.; Chernov, A. A.; Malkin, A.

    2003-01-01

    Canavalin crystals grown from material purified and not purified by High Performance Liquid Chromatography were studied by atomic force microscopy and x-ray diffraction. After purification, resolution was improved from 2.55Angstroms to 2.22Angstroms and jagged isotropic spiral steps transformed into regular, well polygonized steps.

  9. 7 Å Resolution in Protein 2-Dimentional-Crystal X-Ray Diffraction at Linac Coherent Light Source

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pedrini, Bill; Tsai, Ching-Ju; Capitani, Guido

    2014-06-09

    Membrane proteins arranged as two-dimensional (2D) crystals in the lipid en- vironment provide close-to-physiological structural information, which is essential for understanding the molecular mechanisms of protein function. X-ray diffraction from individual 2D crystals did not represent a suitable investigation tool because of radiation damage. The recent availability of ultrashort pulses from X-ray Free Electron Lasers (X-FELs) has now provided a mean to outrun the damage. Here we report on measurements performed at the LCLS X-FEL on bacteriorhodopsin 2D crystals mounted on a solid support and kept at room temperature. By merg- ing data from about a dozen of single crystalmore » diffraction images, we unambiguously identified the diffraction peaks to a resolution of 7 °A, thus improving the observable resolution with respect to that achievable from a single pattern alone. This indicates that a larger dataset will allow for reliable quantification of peak intensities, and in turn a corresponding increase of resolution. The presented results pave the way to further X-FEL studies on 2D crystals, which may include pump-probe experiments at subpicosecond time resolution.« less

  10. Simple fiber-optic confocal microscopy with nanoscale depth resolution beyond the diffraction barrier.

    PubMed

    Ilev, Ilko; Waynant, Ronald; Gannot, Israel; Gandjbakhche, Amir

    2007-09-01

    A novel fiber-optic confocal approach for ultrahigh depth-resolution (

  11. 7 Å resolution in protein two-dimensional-crystal X-ray diffraction at Linac Coherent Light Source

    PubMed Central

    Pedrini, Bill; Tsai, Ching-Ju; Capitani, Guido; Padeste, Celestino; Hunter, Mark S.; Zatsepin, Nadia A.; Barty, Anton; Benner, W. Henry; Boutet, Sébastien; Feld, Geoffrey K.; Hau-Riege, Stefan P.; Kirian, Richard A.; Kupitz, Christopher; Messerschmitt, Marc; Ogren, John I.; Pardini, Tommaso; Segelke, Brent; Williams, Garth J.; Spence, John C. H.; Abela, Rafael; Coleman, Matthew; Evans, James E.; Schertler, Gebhard F. X.; Frank, Matthias; Li, Xiao-Dan

    2014-01-01

    Membrane proteins arranged as two-dimensional crystals in the lipid environment provide close-to-physiological structural information, which is essential for understanding the molecular mechanisms of protein function. Previously, X-ray diffraction from individual two-dimensional crystals did not represent a suitable investigational tool because of radiation damage. The recent availability of ultrashort pulses from X-ray free-electron lasers (XFELs) has now provided a means to outrun the damage. Here, we report on measurements performed at the Linac Coherent Light Source XFEL on bacteriorhodopsin two-dimensional crystals mounted on a solid support and kept at room temperature. By merging data from about a dozen single crystal diffraction images, we unambiguously identified the diffraction peaks to a resolution of 7 Å, thus improving the observable resolution with respect to that achievable from a single pattern alone. This indicates that a larger dataset will allow for reliable quantification of peak intensities, and in turn a corresponding increase in the resolution. The presented results pave the way for further XFEL studies on two-dimensional crystals, which may include pump–probe experiments at subpicosecond time resolution. PMID:24914166

  12. Computer-generated holograms and diffraction gratings in optical security applications

    NASA Astrophysics Data System (ADS)

    Stepien, Pawel J.

    2000-04-01

    The term 'computer generated hologram' (CGH) describes a diffractive structure strictly calculated and recorded to diffract light in a desired way. The CGH surface profile is a result of the wavefront calculation rather than of interference. CGHs are able to form 2D and 3D images. Optically, variable devices (OVDs) composed of diffractive gratings are often used in security applications. There are various types of optically and digitally recorded gratings in security applications. Grating based OVDs are used to record bright 2D images with limited range of cinematic effects. These effects result form various orientations or densities of recorded gratings. It is difficult to record high quality OVDs of 3D objects using gratings. Stereo grams and analogue rainbow holograms offer 3D imaging, but they are darker and have lower resolution than grating OVDs. CGH based OVDs contains unlimited range of cinematic effects and high quality 3D images. Images recorded using CGHs are usually more noisy than grating based OVDs, because of numerical inaccuracies in CGH calculation and mastering. CGH based OVDs enable smooth integration of hidden and machine- readable features within an OVD design.

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

    PubMed Central

    Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin; Lohse, Samuel E.; Lee, Chang-soo; Torelli, Marco D.; Hamers, Robert J.; Murhpy, Catherine J.; Orr, Galya

    2015-01-01

    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 eficient 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. PMID:24816810

  14. Atomistic three-dimensional coherent x-ray imaging of nonbiological systems

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ho, Phay J.; Knight, Chris; Tegze, Miklos

    We computationally study the resolution limits for three-dimensional coherent x-ray diffractive imaging of heavy, nonbiological systems using Ar clusters as a prototype. We treat electronic and nuclear dynamics on an equal footing and remove the frozen-lattice approximation often used in electronic damage studies. We explore the achievable resolution as a function of pulse parameters (fluence level, pulse duration, and photon energy) and particle size. The contribution of combined lattice and electron dynamics is not negligible even for 2 fs pulses, and the Compton scattering is less deleterious than in biological systems for atomic-scale imaging. Although free-electron scattering represents a significantmore » background, we find that recovery of the original structure is in principle possible with 3 angstrom resolution for particles of 11 nm diameter.« less

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin

    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 localizationmore » patterns of nanomaterials, specifically cadmium selenide quantum dots, following interaction with bacterial cells.« less

  16. Atomistic three-dimensional coherent x-ray imaging of nonbiological systems

    DOE PAGES

    Ho, Phay J.; Knight, Chris; Tegze, Miklos; ...

    2016-12-12

    We computationally study the resolution limits for three-dimensional coherent x-ray diffractive imaging of heavy, nonbiological systems using Ar clusters as a prototype. We treat electronic and nuclear dynamics on an equal footing and remove the frozen-lattice approximation often used in electronic damage studies. We explore the achievable resolution as a function of pulse parameters (fluence level, pulse duration, and photon energy) and particle size. The contribution of combined lattice and electron dynamics is not negligible even for 2 fs pulses, and the Compton scattering is less deleterious than in biological systems for atomic-scale imaging. Although free-electron scattering represents a significantmore » background, we find that recovery of the original structure is in principle possible with 3 angstrom resolution for particles of 11 nm diameter.« less

  17. Integrated optics to improve resolution on multiple configuration

    NASA Astrophysics Data System (ADS)

    Liu, Hua; Ding, Quanxin; Guo, Chunjie; Zhou, Liwei

    2015-04-01

    Inspired to in order to reveal the structure to improve imaging resolution, further technical requirement is proposed in some areas of the function and influence on the development of multiple configuration. To breakthrough diffraction limit, smart structures are recommended as the most efficient and economical method, while by used to improve the system performance, especially on signal to noise ratio and resolution. Integrated optics were considered in the selection, with which typical multiple configuration, by use the method of simulation experiment. Methodology can change traditional design concept and to develop the application space. Our calculations using multiple matrix transfer method, also the correlative algorithm and full calculations, show the expected beam shaping through system and, in particular, the experimental results will support our argument, which will be reported in the presentation.

  18. Design and Performance of A High Resolution Micro-Spec: An Integrated Sub-Millimeter Spectrometer

    NASA Technical Reports Server (NTRS)

    Barrentine, Emily M.; Cataldo, Giuseppe; Brown, Ari D.; Ehsan, Negar; Noroozian, Omid; Stevenson, Thomas R.; U-Yen, Kongpop; Wollack, Edward J.; Moseley, S. Harvey

    2016-01-01

    Micro-Spec is a compact sub-millimeter (approximately 100 GHz--1:1 THz) spectrometer which uses low loss superconducting microstrip transmission lines and a single-crystal silicon dielectric to integrate all of the components of a diffraction grating spectrometer onto a single chip. We have already successfully evaluated the performance of a prototype Micro-Spec, with spectral resolving power, R=64. Here we present our progress towards developing a higher resolution Micro-Spec, which would enable the first science returns in a balloon flight version of this instrument. We describe modifications to the design in scaling from a R=64 to a R=256 instrument, as well as the ultimate performance limits and design concerns when scaling this instrument to higher resolutions.

  19. Observing Exoplanetary Ozone In The Mid-Ultraviolet

    NASA Technical Reports Server (NTRS)

    Heap, S.

    2008-01-01

    There are good reasons for pushing the spectral range of observation to shorter wavelengths than currently envisaged for terrestrial planet-finding missions utilizing with a 4-rn, diffraction-limited, optical telescope: (1) The angular resolution is higher, so that the image of an exoplanet is better separated from that of the much brighter star. (2) Due to the higher resolution, the exozodiacal background per resolution element is smaller, so exposure times are reduced for the same incident flux. (3) Most importantly, the sensitivity to the presence of life on habitable exoplanets is increased by a hundred-fold by access to the ozone biomarker at 250-300 nm. These benefits must be weighed against challenges arising from the faintness of exoplanets in the mid-UV. We will describe the benefits and the technical and cost challenges.

  20. Energy resolution of the CdTe-XPAD detector: calibration and potential for Laue diffraction measurements on protein crystals.

    PubMed

    Medjoubi, Kadda; Thompson, Andrew; Bérar, Jean-François; Clemens, Jean-Claude; Delpierre, Pierre; Da Silva, Paulo; Dinkespiler, Bernard; Fourme, Roger; Gourhant, Patrick; Guimaraes, Beatriz; Hustache, Stéphanie; Idir, Mourad; Itié, Jean-Paul; Legrand, Pierre; Menneglier, Claude; Mercere, Pascal; Picca, Frederic; Samama, Jean-Pierre

    2012-05-01

    The XPAD3S-CdTe, a CdTe photon-counting pixel array detector, has been used to measure the energy and the intensity of the white-beam diffraction from a lysozyme crystal. A method was developed to calibrate the detector in terms of energy, allowing incident photon energy measurement to high resolution (approximately 140 eV), opening up new possibilities in energy-resolved X-ray diffraction. In order to demonstrate this, Laue diffraction experiments were performed on the bending-magnet beamline METROLOGIE at Synchrotron SOLEIL. The X-ray energy spectra of diffracted spots were deduced from the indexed Laue patterns collected with an imaging-plate detector and then measured with both the XPAD3S-CdTe and the XPAD3S-Si, a silicon photon-counting pixel array detector. The predicted and measured energy of selected diffraction spots are in good agreement, demonstrating the reliability of the calibration method. These results open up the way to direct unit-cell parameter determination and the measurement of high-quality Laue data even at low resolution. Based on the success of these measurements, potential applications in X-ray diffraction opened up by this type of technology are discussed.

  1. The 2015 super-resolution microscopy roadmap

    NASA Astrophysics Data System (ADS)

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

    2015-11-01

    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 discussion on the concepts underlying super-resolution optical microscopy, the potential of different approaches, the importance of label optimization (such as reversible photoswitchable proteins) and applications in which these methods will have a significant impact. Mark Bates, Christian Eggeling

  2. Hard X-ray multilayer zone plate with 25-nm outermost zone width

    NASA Astrophysics Data System (ADS)

    Takano, H.; Sumida, K.; Hirotomo, H.; Koyama, T.; Ichimaru, S.; Ohchi, T.; Takenaka, H.; Kagoshima, Y.

    2017-06-01

    We have improved the performance of a previously reported multilayer zone plate by reducing its outermost zone width, using the same multilayer materials (MoSi2 and Si) and fabrication technique. The focusing performance was evaluated at the BL24XU of SPring-8 using 20-keV X-rays. The line spread function (LSF) in the focal plane was measured using a dark-field knife-edge scan method, and the point spread function was obtained from the LSF through a tomographic reconstruction principle. The spatial resolution was estimated to be 30 nm, which is in relatively good agreement with the calculated diffraction-limited value of 25 nm, while the measured diffraction efficiency of the +1st order was 24%.

  3. Preliminary neutron diffraction analysis of challenging human manganese superoxide dismutase crystals

    DOE PAGES

    Azadmanesh, Jahaun; Trickel, Scott R.; Weiss, Kevin L.; ...

    2017-03-29

    Superoxide dismutases (SODs) are enzymes that protect against oxidative stress by dismutation of superoxide into oxygen and hydrogen peroxide through cyclic reduction and oxidation of the active-site metal. The complete enzymatic mechanisms of SODs are unknown since data on the positions of hydrogen are limited. Here, we present, methods for large crystal growth and neutron data collection of human manganese SOD (MnSOD) using perdeuteration and the MaNDi beamline at Oak Ridge National Laboratory. Furthermore, The crystal from which the human MnSOD data set was obtained is the crystal with the largest unit-cell edge (240 Å) from which data have beenmore » collectedvianeutron diffraction to sufficient resolution (2.30 Å) where hydrogen positions can be observed.« less

  4. Sub-diffraction Laser Synthesis of Silicon Nanowires

    PubMed Central

    Mitchell, James I.; Zhou, Nan; Nam, Woongsik; Traverso, Luis M.; Xu, Xianfan

    2014-01-01

    We demonstrate synthesis of silicon nanowires of tens of nanometers via laser induced chemical vapor deposition. These nanowires with diameters as small as 60 nm are produced by the interference between incident laser radiation and surface scattered radiation within a diffraction limited spot, which causes spatially confined, periodic heating needed for high resolution chemical vapor deposition. By controlling the intensity and polarization direction of the incident radiation, multiple parallel nanowires can be simultaneously synthesized. The nanowires are produced on a dielectric substrate with controlled diameter, length, orientation, and the possibility of in-situ doping, and therefore are ready for device fabrication. Our method offers rapid one-step fabrication of nano-materials and devices unobtainable with previous CVD methods. PMID:24469704

  5. Preliminary neutron diffraction analysis of challenging human manganese superoxide dismutase crystals

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Azadmanesh, Jahaun; Trickel, Scott R.; Weiss, Kevin L.

    Superoxide dismutases (SODs) are enzymes that protect against oxidative stress by dismutation of superoxide into oxygen and hydrogen peroxide through cyclic reduction and oxidation of the active-site metal. The complete enzymatic mechanisms of SODs are unknown since data on the positions of hydrogen are limited. Here, we present, methods for large crystal growth and neutron data collection of human manganese SOD (MnSOD) using perdeuteration and the MaNDi beamline at Oak Ridge National Laboratory. Furthermore, The crystal from which the human MnSOD data set was obtained is the crystal with the largest unit-cell edge (240 Å) from which data have beenmore » collectedvianeutron diffraction to sufficient resolution (2.30 Å) where hydrogen positions can be observed.« less

  6. One-dimensional error-diffusion technique adapted for binarization of rotationally symmetric pupil filters

    NASA Astrophysics Data System (ADS)

    Kowalczyk, Marek; Martínez-Corral, Manuel; Cichocki, Tomasz; Andrés, Pedro

    1995-02-01

    Two novel algorithms for the binarization of continuous rotationally symmetric real and positive pupil filters are presented. Both algorithms are based on the one-dimensional error diffusion concept. In our numerical experiment an original gray-tone apodizer is substituted by a set of transparent and opaque concentric annular zones. Depending on the algorithm the resulting binary mask consists of either equal width or equal area zones. The diffractive behavior of binary filters is evaluated. It is shown that the filter with equal width zones gives Fraunhofer diffraction pattern more similar to that of the original gray-tone apodizer than that with equal area zones, assuming in both cases the same resolution limit of device used to print both filters.

  7. Revealing the Effects of Nanoscale Membrane Curvature on Lipid Mobility.

    PubMed

    Kabbani, Abir Maarouf; Woodward, Xinxin; Kelly, Christopher V

    2017-10-18

    Recent advances in nanoengineering and super-resolution microscopy have enabled new capabilities for creating and observing membrane curvature. However, the effects of curvature on single-lipid diffusion have yet to be revealed. The simulations presented here describe the capabilities of varying experimental methods for revealing the effects of nanoscale curvature on single-molecule mobility. Traditionally, lipid mobility is revealed through fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), and single particle tracking (SPT). However, these techniques vary greatly in their ability to detect the effects of nanoscale curvature on lipid behavior. Traditionally, FRAP and FCS depend on diffraction-limited illumination and detection. A simulation of FRAP shows minimal effects on lipids diffusion due to a 50 nm radius membrane bud. Throughout the stages of the budding process, FRAP detected minimal changes in lipid recovery time due to the curvature versus flat membrane. Simulated FCS demonstrated small effects due to a 50 nm radius membrane bud that was more apparent with curvature-dependent lipid mobility changes. However, SPT achieves a sub-diffraction-limited resolution of membrane budding and lipid mobility through the identification of the single-lipid positions with ≤15 nm spatial and ≤20 ms temporal resolution. By mapping the single-lipid step lengths to locations on the membrane, the effects of membrane topography and curvature could be correlated to the effective membrane viscosity. Single-fluorophore localization techniques, such SPT, can detect membrane curvature and its effects on lipid behavior. These simulations and discussion provide a guideline for optimizing the experimental procedures in revealing the effects of curvature on lipid mobility and effective local membrane viscosity.

  8. Investigating radiation induced damage processes with femtosecond x-ray pulses (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Song, Changyong

    2017-05-01

    Interest in high-resolution structure investigation has been zealous, especially with the advent of X-ray free electron lasers (XFELs). The intense and ultra-short X-ray laser pulses ( 10 GW) pave new routes to explore structures and dynamics of single macromolecules, functional nanomaterials and complex electronic materials. In the last several years, we have developed XFEL single-shot diffraction imaging by probing ultrafast phase changes directly. Pump-probe single-shot imaging was realized by synchronizing femtosecond (<10 fs in FWHM) X-ray laser (probe) with femtosecond (50 fs) IR laser (pump) at better than 1 ps resolution. Nanoparticles under intense fs-laser pulses were investigated with fs XFEL pulses to provide insight into the irreversible particle damage processes with nanoscale resolution. Research effort, introduced, aims to extend the current spatio-temporal resolution beyond the present limit. We expect this single-shot dynamic imaging to open new science opportunity with XFELs.

  9. Quantitative super-resolution imaging of Bruchpilot distinguishes active zone states

    NASA Astrophysics Data System (ADS)

    Ehmann, Nadine; van de Linde, Sebastian; Alon, Amit; Ljaschenko, Dmitrij; Keung, Xi Zhen; Holm, Thorge; Rings, Annika; Diantonio, Aaron; Hallermann, Stefan; Ashery, Uri; Heckmann, Manfred; Sauer, Markus; Kittel, Robert J.

    2014-08-01

    The precise molecular architecture of synaptic active zones (AZs) gives rise to different structural and functional AZ states that fundamentally shape chemical neurotransmission. However, elucidating the nanoscopic protein arrangement at AZs is impeded by the diffraction-limited resolution of conventional light microscopy. Here we introduce new approaches to quantify endogenous protein organization at single-molecule resolution in situ with super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM). Focusing on the Drosophila neuromuscular junction (NMJ), we find that the AZ cytomatrix (CAZ) is composed of units containing ~137 Bruchpilot (Brp) proteins, three quarters of which are organized into about 15 heptameric clusters. We test for a quantitative relationship between CAZ ultrastructure and neurotransmitter release properties by engaging Drosophila mutants and electrophysiology. Our results indicate that the precise nanoscopic organization of Brp distinguishes different physiological AZ states and link functional diversification to a heretofore unrecognized neuronal gradient of the CAZ ultrastructure.

  10. High resolution X-ray diffraction imaging of lead tin telluride

    NASA Technical Reports Server (NTRS)

    Steiner, Bruce; Dobbyn, Ronald C.; Black, David; Burdette, Harold; Kuriyama, Masao; Spal, Richard; Simchick, Richard; Fripp, Archibald

    1991-01-01

    High resolution X-ray diffraction images of two directly comparable crystals of lead tin telluride, one Bridgman-grown on Space Shuttle STS 61A and the other terrestrially Bridgman-grown under similar conditions from identical material, present different subgrain structure. In the terrestrial, sample 1 the appearance of an elaborate array of subgrains is closely associated with the intrusion of regions that are out of diffraction in all of the various images. The formation of this elaborate subgrain structure is inhibited by growth in microgravity.

  11. Variable Magnification With Kirkpatrick-Baez Optics for Synchrotron X-Ray Microscopy

    PubMed Central

    Jach, Terrence; Bakulin, Alex S.; Durbin, Stephen M.; Pedulla, Joseph; Macrander, Albert

    2006-01-01

    We describe the distinction between the operation of a short focal length x-ray microscope forming a real image with a laboratory source (convergent illumination) and with a highly collimated intense beam from a synchrotron light source (Köhler illumination). We demonstrate the distinction with a Kirkpatrick-Baez microscope consisting of short focal length multilayer mirrors operating at an energy of 8 keV. In addition to realizing improvements in the resolution of the optics, the synchrotron radiation microscope is not limited to the usual single magnification at a fixed image plane. Higher magnification images are produced by projection in the limit of geometrical optics with a collimated beam. However, in distinction to the common method of placing the sample behind the optical source of a diverging beam, we describe the situation in which the sample is located in the collimated beam before the optical element. The ultimate limits of this magnification result from diffraction by the specimen and are determined by the sample position relative to the focal point of the optic. We present criteria by which the diffraction is minimized. PMID:27274930

  12. Kernel-Phase Interferometry for Super-Resolution Detection of Faint Companions

    NASA Astrophysics Data System (ADS)

    Factor, Samuel M.; Kraus, Adam L.

    2017-01-01

    Direct detection of close in companions (exoplanets or binary systems) is notoriously difficult. While coronagraphs and point spread function (PSF) subtraction can be used to reduce contrast and dig out signals of companions under the PSF, there are still significant limitations in separation and contrast. Non-redundant aperture masking (NRM) interferometry can be used to detect companions well inside the PSF of a diffraction limited image, though the mask discards ˜95% of the light gathered by the telescope and thus the technique is severely flux limited. Kernel-phase analysis applies interferometric techniques similar to NRM to a diffraction limited image utilizing the full aperture. Instead of non-redundant closure-phases, kernel-phases are constructed from a grid of points on the full aperture, simulating a redundant interferometer. I have developed my own faint companion detection pipeline which utilizes an Bayesian analysis of kernel-phases. I have used this pipeline to search for new companions in archival images from HST/NICMOS in order to constrain planet and binary formation models at separations inaccessible to previous techniques. Using this method, it is possible to detect a companion well within the classical λ/D Rayleigh diffraction limit using a fraction of the telescope time as NRM. This technique can easily be applied to archival data as no mask is needed and will thus make the detection of close in companions cheap and simple as no additional observations are needed. Since the James Webb Space Telescope (JWST) will be able to perform NRM observations, further development and characterization of kernel-phase analysis will allow efficient use of highly competitive JWST telescope time.

  13. Pupil-segmentation-based adaptive optical correction of a high-numerical-aperture gradient refractive index lens for two-photon fluorescence endoscopy.

    PubMed

    Wang, Chen; Ji, Na

    2012-06-01

    The intrinsic aberrations of high-NA gradient refractive index (GRIN) lenses limit their image quality as well as field of view. Here we used a pupil-segmentation-based adaptive optical approach to correct the inherent aberrations in a two-photon fluorescence endoscope utilizing a 0.8 NA GRIN lens. By correcting the field-dependent aberrations, we recovered diffraction-limited performance across a large imaging field. The consequent improvements in imaging signal and resolution allowed us to detect fine structures that were otherwise invisible inside mouse brain slices.

  14. Observation of femtosecond X-ray interactions with matter using an X-ray–X-ray pump–probe scheme

    PubMed Central

    Inoue, Ichiro; Inubushi, Yuichi; Sato, Takahiro; Tono, Kensuke; Katayama, Tetsuo; Kameshima, Takashi; Ogawa, Kanade; Togashi, Tadashi; Owada, Shigeki; Amemiya, Yoshiyuki; Tanaka, Takashi; Hara, Toru

    2016-01-01

    Resolution in the X-ray structure determination of noncrystalline samples has been limited to several tens of nanometers, because deep X-ray irradiation required for enhanced resolution causes radiation damage to samples. However, theoretical studies predict that the femtosecond (fs) durations of X-ray free-electron laser (XFEL) pulses make it possible to record scattering signals before the initiation of X-ray damage processes; thus, an ultraintense X-ray beam can be used beyond the conventional limit of radiation dose. Here, we verify this scenario by directly observing femtosecond X-ray damage processes in diamond irradiated with extraordinarily intense (∼1019 W/cm2) XFEL pulses. An X-ray pump–probe diffraction scheme was developed in this study; tightly focused double–5-fs XFEL pulses with time separations ranging from sub-fs to 80 fs were used to excite (i.e., pump) the diamond and characterize (i.e., probe) the temporal changes of the crystalline structures through Bragg reflection. It was found that the pump and probe diffraction intensities remain almost constant for shorter time separations of the double pulse, whereas the probe diffraction intensities decreased after 20 fs following pump pulse irradiation due to the X-ray–induced atomic displacement. This result indicates that sub-10-fs XFEL pulses enable conductions of damageless structural determinations and supports the validity of the theoretical predictions of ultraintense X-ray–matter interactions. The X-ray pump–probe scheme demonstrated here would be effective for understanding ultraintense X-ray–matter interactions, which will greatly stimulate advanced XFEL applications, such as atomic structure determination of a single molecule and generation of exotic matters with high energy densities. PMID:26811449

  15. The High Resolution Powder Diffraction Beam Line at ESRF.

    PubMed

    Fitch, A N

    2004-01-01

    The optical design and performance of the high-resolution powder diffraction beam line BM16 at ESRF are discussed and illustrated. Some recent studies carried out on BM16 are described, including crystal structure solution and refinement, anomalous scattering, in situ measurements, residual strain in engineering components, investigation of microstructure, and grazing-incidence diffraction from surface layers. The beam line is built on a bending magnet, and operates in the energy range from 5 keV to 40 keV. After the move to an undulator source in 2002, it will benefit from an extented energy range up to 60 keV and increased flux and resolution. It is anticipated that enhancements to the data quality will be achieved, leading to the solution of larger crystal structures, and improvements in the accuracy of refined structures. The systematic exploitation of anisotropic thermal expansion will help reduce the effects of peak overlap in the analysis of powder diffraction data.

  16. Crystallization of a fungal lytic polysaccharide monooxygenase expressed from glycoengineered Pichia pastoris for X-ray and neutron diffraction

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    O'Dell, William B.; Swartz, Paul D.; Weiss, Kevin L.

    Lytic polysaccharide monooxygenases (LPMOs) are carbohydrate-disrupting enzymes secreted by bacteria and fungi that break glycosidic bondsviaan oxidative mechanism. Fungal LPMOs typically act on cellulose and can enhance the efficiency of cellulose-hydrolyzing enzymes that release soluble sugars for bioethanol production or other industrial uses. The enzyme PMO-2 fromNeurospora crassa(NcPMO-2) was heterologously expressed inPichia pastoristo facilitate crystallographic studies of the fungal LPMO mechanism. Diffraction resolution and crystal morphology were improved by expressingNcPMO-2 from a glycoengineered strain ofP. pastorisand by the use of crystal seeding methods, respectively. These improvements resulted in high-resolution (1.20 Å) X-ray diffraction data collection at 100 K and themore » production of a largeNcPMO-2 crystal suitable for room-temperature neutron diffraction data collection to 2.12 Å resolution.« less

  17. Understanding deformation with high angular resolution electron backscatter diffraction (HR-EBSD)

    NASA Astrophysics Data System (ADS)

    Britton, T. B.; Hickey, J. L. R.

    2018-01-01

    High angular resolution electron backscatter diffraction (HR-EBSD) affords an increase in angular resolution, as compared to ‘conventional’ Hough transform based EBSD, of two orders of magnitude, enabling measurements of relative misorientations of 1 x 10-4 rads (~ 0.006°) and changes in (deviatoric) lattice strain with a precision of 1 x 10-4. This is achieved through direct comparison of two or more diffraction patterns using sophisticated cross-correlation based image analysis routines. Image shifts between zone axes in the two-correlated diffraction pattern are measured with sub-pixel precision and this realises the ability to measure changes in interplanar angles and lattice orientation with a high degree of sensitivity. These shifts are linked to strains and lattice rotations through simple geometry. In this manuscript, we outline the basis of the technique and two case studies that highlight its potential to tackle real materials science challenges, such as deformation patterning in polycrystalline alloys.

  18. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto.

    PubMed

    Yanagisawa, Yasuhide; Chatake, Toshiyuki; Chiba-Kamoshida, Kaori; Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki; Yasuda, Ichiro; Morimoto, Yukio

    2010-12-01

    Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27,724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a=74.3, b=49.9, c=56.3 Å, β=95.2°. Diffraction images were processed to a resolution of 1.74 Å with an Rmerge of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase.

  19. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto

    PubMed Central

    Yanagisawa, Yasuhide; Chatake, Toshiyuki; Chiba-Kamoshida, Kaori; Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki; Yasuda, Ichiro; Morimoto, Yukio

    2010-01-01

    Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27 724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a = 74.3, b = 49.9, c = 56.3 Å, β = 95.2°. Diffraction images were processed to a resolution of 1.74 Å with an R merge of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase. PMID:21139221

  20. Crystallization and preliminary X-ray diffraction analysis of the wild-type haloalkane dehalogenase DhaA and its variant DhaA13 complexed with different ligands.

    PubMed

    Stsiapanava, Alena; Chaloupkova, Radka; Fortova, Andrea; Brynda, Jiri; Weiss, Manfred S; Damborsky, Jiri; Smatanova, Ivana Kuta

    2011-02-01

    Haloalkane dehalogenases make up an important class of hydrolytic enzymes which catalyse the cleavage of carbon-halogen bonds in halogenated aliphatic compounds. There is growing interest in these enzymes owing to their potential use in environmental and industrial applications. The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 can slowly detoxify the industrial pollutant 1,2,3-trichloropropane (TCP). Structural analysis of this enzyme complexed with target ligands was conducted in order to obtain detailed information about the structural limitations of its catalytic properties. In this study, the crystallization and preliminary X-ray analysis of complexes of wild-type DhaA with 2-propanol and with TCP and of complexes of the catalytically inactive variant DhaA13 with the dye coumarin and with TCP are described. The crystals of wild-type DhaA were plate-shaped and belonged to the triclinic space group P1, while the variant DhaA13 can form prism-shaped crystals belonging to the orthorhombic space group P2(1)2(1)2(1) as well as plate-shaped crystals belonging to the triclinic space group P1. Diffraction data for crystals of wild-type DhaA grown from crystallization solutions with different concentrations of 2-propanol were collected to 1.70 and 1.26 Å resolution, respectively. A prism-shaped crystal of DhaA13 complexed with TCP and a plate-shaped crystal of the same variant complexed with the dye coumarin diffracted X-rays to 1.60 and 1.33 Å resolution, respectively. A crystal of wild-type DhaA and a plate-shaped crystal of DhaA13, both complexed with TCP, diffracted to atomic resolutions of 1.04 and 0.97 Å, respectively.

  1. A dedicated superbend x-ray microdiffraction beamline for materials, geo-, and environmental sciences at the advanced light source.

    PubMed

    Kunz, Martin; Tamura, Nobumichi; Chen, Kai; MacDowell, Alastair A; Celestre, Richard S; Church, Matthew M; Fakra, Sirine; Domning, Edward E; Glossinger, James M; Kirschman, Jonathan L; Morrison, Gregory Y; Plate, Dave W; Smith, Brian V; Warwick, Tony; Yashchuk, Valeriy V; Padmore, Howard A; Ustundag, Ersan

    2009-03-01

    A new facility for microdiffraction strain measurements and microfluorescence mapping has been built on beamline 12.3.2 at the advanced light source of the Lawrence Berkeley National Laboratory. This beamline benefits from the hard x-radiation generated by a 6 T superconducting bending magnet (superbend). This provides a hard x-ray spectrum from 5 to 22 keV and a flux within a 1 microm spot of approximately 5x10(9) photons/s (0.1% bandwidth at 8 keV). The radiation is relayed from the superbend source to a focus in the experimental hutch by a toroidal mirror. The focus spot is tailored by two pairs of adjustable slits, which serve as secondary source point. Inside the lead hutch, a pair of Kirkpatrick-Baez (KB) mirrors placed in a vacuum tank refocuses the secondary slit source onto the sample position. A new KB-bending mechanism with active temperature stabilization allows for more reproducible and stable mirror bending and thus mirror focusing. Focus spots around 1 microm are routinely achieved and allow a variety of experiments, which have in common the need of spatial resolution. The effective spatial resolution (approximately 0.2 microm) is limited by a convolution of beam size, scan-stage resolution, and stage stability. A four-bounce monochromator consisting of two channel-cut Si(111) crystals placed between the secondary source and KB-mirrors allows for easy changes between white-beam and monochromatic experiments while maintaining a fixed beam position. High resolution stage scans are performed while recording a fluorescence emission signal or an x-ray diffraction signal coming from either a monochromatic or a white focused beam. The former allows for elemental mapping, whereas the latter is used to produce two-dimensional maps of crystal-phases, -orientation, -texture, and -strain/stress. Typically achieved strain resolution is in the order of 5x10(-5) strain units. Accurate sample positioning in the x-ray focus spot is achieved with a commercial laser-triangulation unit. A Si-drift detector serves as a high-energy-resolution (approximately 150 eV full width at half maximum) fluorescence detector. Fluorescence scans can be collected in continuous scan mode with up to 300 pixels/s scan speed. A charge coupled device area detector is utilized as diffraction detector. Diffraction can be performed in reflecting or transmitting geometry. Diffraction data are processed using XMAS, an in-house written software package for Laue and monochromatic microdiffraction analysis.

  2. Superresolution Imaging with Standard Fluorescent Probes

    PubMed Central

    Burnette, Dylan T.; Lippincott-Schwartz, Jennifer; Kachar, Bechara

    2013-01-01

    For more than 100 years, the ultimate resolution of a light microscope (~200 nm) has been constrained by the fundamental physical phenomenon of diffraction, as described by Ernst Abbe in 1873. While this limitation is just as applicable to today’s light microscopes, it is the combination of high-end optics, clever methods of sample illumination, and computational techniques that has enabled researchers to access high-resolution information an order of magnitude greater than once thought possible. This combination, broadly termed superresolution microscopy, has been increasingly practical for many labs to implement from both a hardware and software standpoint, but as with many cutting-edge techniques, it also comes with limitations. One of the current drawbacks to superresolution microscopy is the limited number of probes and conditions that have been suitable for imaging. Here, a technique termed bleaching/blinking assisted localization microscopy (BaLM) makes use of almost all fluorophore’s inherent blinking and bleaching properties as a means to generate superresolution images. PMID:24510788

  3. Overcoming a hemihedral twinning problem in tetrahydrofolate-dependent O -demethylase crystals by the microseeding method

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Harada, Ayaka; Sato, Yukari; Kamimura, Naofumi

    2016-11-30

    A tetrahydrofolate-dependentO-demethylase, LigM, from Sphingobiumsp. SYK-6 was crystallized by the hanging-drop vapour-diffusion method. However, the obtained P3 121 orP3 221 crystals, which diffracted to 2.5–3.3 Å resolution, were hemihedrally twinned. To overcome the twinning problem, microseeding using P3 121/P3 2 21 crystals as microseeds was performed with optimization of the reservoir conditions. As a result, another crystal form was obtained. The newly obtained crystal diffracted to 2.5–3.0 Å resolution and belonged to space group P2 12 12, with unit-cell parametersa= 102.0,b= 117.3,c = 128.1 Å. The P2 12 12 crystals diffracted to better than 2.0 Å resolution after optimizing themore » cryoconditions. Phasing using the single anomalous diffraction method was successful at 3.0 Å resolution with a Pt-derivative crystal. This experience suggested that microseeding is an effective method to overcome the twinning problem, even when twinned crystals are utilized as microseeds.« less

  4. High Sensitivity Detection of Broadband Acoustic Vibration Using Optical Demodulation Method

    NASA Astrophysics Data System (ADS)

    Zhang, Zhen

    Measuring the high frequency acoustic vibrations represents the fundamental interest in revealing the intrinsic dynamic characteristic of board range of systems, such as the growth of the fetus, blood flow in human palms, and vibrations of carbon nanotube. However, the acoustic wave detection capability is limited by the detection bandwidth and sensitivity of the commonly used piezoelectric based ultrasound detectors. To overcome these limitations, this thesis focuses on exploring the optical demodulation method for highly sensitive detection of broadband acoustic vibration. First, a transparent optical ultrasonic detector has been developed using micro-ring resonator (MRR) made of soft polymeric materials. It outperforms the traditional piezoelectric detectors with broader detection bandwidth, miniaturized size and wide angular sensitivity. Its ease of integration into photoacoustic microscopy system has resulted in the great improvement of the imaging resolution. A theoretic framework has been developed to establish the quantitative understanding of its unique distance and angular dependent detection characteristics and was subsequently validated experimentally. The developed theoretic framework provides a guideline to fully accounts for the trade-offs between axial and lateral resolution, working distance, and the field of view in developing optimal imaging performance for a wide range of biological and clinical applications. MRR-based ultrasonic detector is further integrated into confocal fluorescence microscopy to realize the simultaneous imaging of fluorescence and optical absorption of retinal pigment epithelium, achieving multi-contrast imaging at sub-cellular level. The needs to resolve the fine details of the biological specimen with the resolution beyond the diffraction limit further motivate the development of optical demodulated ultrasonic detection method based on near-field scanning optical microscopy (NSOM). The nano-focusing probe was developed for adiabatic focusing of surface plasmon polaritons to the probe apex with high energy efficiency and the suppression of the background noise was accomplished through the implementation of the harmonic demodulation technique. Collectively, this system is capable of delivering intense near-field illumination source while effectively suppressing the background signal due to the far-field scattering and thus, allows for quantitative mapping of local evanescent field with enhanced contrast and improved resolutions. The performance of the developed NSOM system has been validated through the experimental measurements of the surface plasmon polariton mode. This new NSOM system enables optical demodulated ultrasound detection at nanoscale spatial resolution. Using it to detect the ultrasound signal within the acoustic near-field has led to the successful experimental demonstration of the sub-surface photoacoustic imaging of buried objects with sub-diffraction-limited resolution and high sensitivity. Such a new ultrasound detection method holds promising potential for super-resolution ultrasound imaging.

  5. Development of a Tip-Enhanced Near-Field Optical Microscope for Nanoscale Interrogation of Surface Chemistry and Plasmonic Phenomena

    NASA Astrophysics Data System (ADS)

    Heilman, Alexander Lee

    Optical microscopy and spectroscopy are invaluable tools for the physical and chemical characterization of materials and surfaces in a wide range of scientific disciplines. However, the application of conventional optical methods in the study of nanomaterials is inherently limited by diffraction. Tip-enhanced near-field optical microscopy (TENOM) is a hybrid technique that marries optical spectroscopy with scanning probe microscopy to overcome the spatial resolution limit imposed by diffraction. By coupling optical energy into the plasmonic modes of a sharp metal probe tip, a strong, localized optical field is generated near the tip's apex and is used to enhance spectroscopic emissions within a sub-diffraction-limited volume. In this thesis, we describe the design, construction, validation, and application of a custom TENOM instrument with a unique attenuated total reflectance (ATR)-geometry excitation/detection system. The specific goals of this work were: (i) to develop a versatile TENOM instrument capable of investigating a variety of optical phenomena at the nanoscale, (ii) to use the instrument to demonstrate chemical interrogation of surfaces with sub-diffraction-limited spatial resolution (i.e., at super resolution), (iii) to apply the instrument to study plasmonic phenomena that influence spectroscopic enhancement in TENOM measurements, and (iv) to leverage resulting insights to develop systematic improvements that expand the ultimate capabilities of near-field optical interrogation techniques. The TENOM instrument described herein is comprised of three main components: an atomic force microscope (AFM), a side-on confocal Raman microscope, and a novel ATR excitation/detection system. The design of each component is discussed along with the results of relevant validation experiments, which were performed to rigorously assess each component's performance. Finite-difference time-domain (FDTD) optical simulations were also developed and used extensively to evaluate the results of validation studies and to optimize experimental design and instrument performance. By combining and synchronizing the operation of the instrument's three components, we perform a variety of near-field optical experiments that demonstrate the instrument's functionality and versatility. ATR illumination is combined with a plasmonic AFM tip to show that: (i) the tip can quantitatively transduce the optical near-field (evanescent waves) above the surface by scattering photons into the far-field, (ii) the ATR geometry enables excitation and characterization of surface plasmon polaritons (SPPs), whose associated optical fields are shown to enhance Raman scattering from a thin layer of copper phthalocyanine (CuPc), and (iii) SPPs can be used to plasmonically excite the tip for super-resolution chemical imaging of patterned CuPc via tip-enhanced Raman spectroscopy (TERS). ATR-illumination TERS is quantitatively compared with side-on illumination. In both cases, spatial resolution was better than 40 nm and tip-on/tip-off Raman enhancement factors were >6500. Furthermore, ATR illumination was shown to provide similar Raman signal levels at lower "effective'' pump powers due to additional optical energy delivered by SPPs to the active region in the tip-surface gap. We also investigate the sensitivity of the TENOM instrument to changes in the plasmonic properties of the tip-surface system in the strongly-coupled regime at small tip-surface separations. Specifically, we demonstrate detection of a resonant plasmonic tip-surface mode (a gap plasmon) that dramatically influences the optical response of the system, and we use experimental results and FDTD simulations to support a hypothesized mechanism. Moreover, we confirm that the gap plasmon resonance has a strong effect on the enhancement of both fluorescence and Raman scattering, and we propose that this phenomenon could ultimately be exploited to improve sensitivity in super-resolution chemical imaging measurements. Finally, we recommend a straightforward modification to the TENOM instrument that could enable future application of these gap-mode plasmon resonances to increase spectroscopic enhancements by an order of magnitude.

  6. xMDFF: molecular dynamics flexible fitting of low-resolution X-ray structures.

    PubMed

    McGreevy, Ryan; Singharoy, Abhishek; Li, Qufei; Zhang, Jingfen; Xu, Dong; Perozo, Eduardo; Schulten, Klaus

    2014-09-01

    X-ray crystallography remains the most dominant method for solving atomic structures. However, for relatively large systems, the availability of only medium-to-low-resolution diffraction data often limits the determination of all-atom details. A new molecular dynamics flexible fitting (MDFF)-based approach, xMDFF, for determining structures from such low-resolution crystallographic data is reported. xMDFF employs a real-space refinement scheme that flexibly fits atomic models into an iteratively updating electron-density map. It addresses significant large-scale deformations of the initial model to fit the low-resolution density, as tested with synthetic low-resolution maps of D-ribose-binding protein. xMDFF has been successfully applied to re-refine six low-resolution protein structures of varying sizes that had already been submitted to the Protein Data Bank. Finally, via systematic refinement of a series of data from 3.6 to 7 Å resolution, xMDFF refinements together with electrophysiology experiments were used to validate the first all-atom structure of the voltage-sensing protein Ci-VSP.

  7. Analytical description of high-aperture STED resolution with 0–2π vortex phase modulation

    PubMed Central

    Xie, Hao; Liu, Yujia; Jin, Dayong; Santangelo, Philip J.; Xi, Peng

    2014-01-01

    Stimulated emission depletion (STED) can achieve optical superresolution, with the optical diffraction limit broken by the suppression on the periphery of the fluorescent focal spot. Previously, it is generally experimentally accepted that there exists an inverse square root relationship with the STED power and the resolution, but with arbitrary coefficients in expression. In this paper, we have removed the arbitrary coefficients by exploring the relationship between the STED power and the achievable resolution from vector optical theory for the widely used 0–2π vortex phase modulation. Electromagnetic fields of the focal region of a high numerical aperture objective are calculated and approximated into polynomials of radius in the focal plane, and analytical expression of resolution as a function of the STED intensity has been derived. As a result, the resolution can be estimated directly from the measurement of the saturation power of the dye and the STED power applied in the region of high STED power. PMID:24323224

  8. Plasmon-emitter interaction using integrated ring grating-nanoantenna structures.

    PubMed

    Rahbany, Nancy; Geng, Wei; Bachelot, Renaud; Couteau, Christophe

    2017-05-05

    Overcoming the diffraction limit to achieve high optical resolution is one of the main challenges in the fields of plasmonics, nanooptics and nanophotonics. In this work, we introduce novel plasmonic structures consisting of nanoantennas (nanoprisms, single bowtie nanoantennas and double bowtie nanoantennas) integrated in the center of ring diffraction gratings. Propagating surface plasmon polaritons (SPPs) are generated by the ring grating and coupled with localized surface plasmons (LSPs) at the nanoantennas exciting emitters placed in their gap. SPPs are widely used for optical waveguiding but provide low resolution due to their weak spatial confinement. In contrast, LSPs provide excellent sub-wavelength confinement but induce large losses. The phenomenon of SPP-LSP coupling witnessed in our structures allows for achieving more precise focusing at the nanoscale, causing an increase in the fluorescence emission of the emitters. Finite-difference time-domain simulations as well as experimental fabrication and optical characterization results are presented to study plasmon-emitter coupling between an ensemble of dye molecules and our integrated plasmonic structures. A comparison is given to highlight the importance of each structure on the photoluminescence and radiative decay enhancement of the molecules.

  9. Plasmon-emitter interaction using integrated ring grating-nanoantenna structures

    NASA Astrophysics Data System (ADS)

    Rahbany, Nancy; Geng, Wei; Bachelot, Renaud; Couteau, Christophe

    2017-05-01

    Overcoming the diffraction limit to achieve high optical resolution is one of the main challenges in the fields of plasmonics, nanooptics and nanophotonics. In this work, we introduce novel plasmonic structures consisting of nanoantennas (nanoprisms, single bowtie nanoantennas and double bowtie nanoantennas) integrated in the center of ring diffraction gratings. Propagating surface plasmon polaritons (SPPs) are generated by the ring grating and coupled with localized surface plasmons (LSPs) at the nanoantennas exciting emitters placed in their gap. SPPs are widely used for optical waveguiding but provide low resolution due to their weak spatial confinement. In contrast, LSPs provide excellent sub-wavelength confinement but induce large losses. The phenomenon of SPP-LSP coupling witnessed in our structures allows for achieving more precise focusing at the nanoscale, causing an increase in the fluorescence emission of the emitters. Finite-difference time-domain simulations as well as experimental fabrication and optical characterization results are presented to study plasmon-emitter coupling between an ensemble of dye molecules and our integrated plasmonic structures. A comparison is given to highlight the importance of each structure on the photoluminescence and radiative decay enhancement of the molecules.

  10. New software to model energy dispersive X-ray diffraction in polycrystalline materials

    NASA Astrophysics Data System (ADS)

    Ghammraoui, B.; Tabary, J.; Pouget, S.; Paulus, C.; Moulin, V.; Verger, L.; Duvauchelle, Ph.

    2012-02-01

    Detection of illicit materials, such as explosives or drugs, within mixed samples is a major issue, both for general security and as part of forensic analyses. In this paper, we describe a new code simulating energy dispersive X-ray diffraction patterns in polycrystalline materials. This program, SinFullscat, models diffraction of any object in any diffractometer system taking all physical phenomena, including amorphous background, into account. Many system parameters can be tuned: geometry, collimators (slit and cylindrical), sample properties, X-ray source and detector energy resolution. Good agreement between simulations and experimental data was obtained. Simulations using explosive materials indicated that parameters such as the diffraction angle or the energy resolution of the detector have a significant impact on the diffraction signature of the material inspected. This software will be a convenient tool to test many diffractometer configurations, providing information on the one that best restores the spectral diffraction signature of the materials of interest.

  11. A possibility of parallel and anti-parallel diffraction measurements on neu- tron diffractometer employing bent perfect crystal monochromator at the monochromatic focusing condition

    NASA Astrophysics Data System (ADS)

    Choi, Yong Nam; Kim, Shin Ae; Kim, Sung Kyu; Kim, Sung Baek; Lee, Chang-Hee; Mikula, Pavel

    2004-07-01

    In a conventional diffractometer having single monochromator, only one position, parallel position, is used for the diffraction experiment (i.e. detection) because the resolution property of the other one, anti-parallel position, is very poor. However, a bent perfect crystal (BPC) monochromator at monochromatic focusing condition can provide a quite flat and equal resolution property at both parallel and anti-parallel positions and thus one can have a chance to use both sides for the diffraction experiment. From the data of the FWHM and the Delta d/d measured on three diffraction geometries (symmetric, asymmetric compression and asymmetric expansion), we can conclude that the simultaneous diffraction measurement in both parallel and anti-parallel positions can be achieved.

  12. Neutron and X-ray single-crystal diffraction from protein microcrystals via magnetically oriented microcrystal arrays in gels.

    PubMed

    Tsukui, Shu; Kimura, Fumiko; Kusaka, Katsuhiro; Baba, Seiki; Mizuno, Nobuhiro; Kimura, Tsunehisa

    2016-07-01

    Protein microcrystals magnetically aligned in D2O hydrogels were subjected to neutron diffraction measurements, and reflections were observed for the first time to a resolution of 3.4 Å from lysozyme microcrystals (∼10 × 10 × 50 µm). This result demonstrated the possibility that magnetically oriented microcrystals consolidated in D2O gels may provide a promising means to obtain single-crystal neutron diffraction from proteins that do not crystallize at the sizes required for neutron diffraction structure determination. In addition, lysozyme microcrystals aligned in H2O hydrogels allowed structure determination at a resolution of 1.76 Å at room temperature by X-ray diffraction. The use of gels has advantages since the microcrystals are measured under hydrated conditions.

  13. Lens-based wavefront sensorless adaptive optics swept source OCT

    NASA Astrophysics Data System (ADS)

    Jian, Yifan; Lee, Sujin; Ju, Myeong Jin; Heisler, Morgan; Ding, Weiguang; Zawadzki, Robert J.; Bonora, Stefano; Sarunic, Marinko V.

    2016-06-01

    Optical coherence tomography (OCT) has revolutionized modern ophthalmology, providing depth resolved images of the retinal layers in a system that is suited to a clinical environment. Although the axial resolution of OCT system, which is a function of the light source bandwidth, is sufficient to resolve retinal features at a micrometer scale, the lateral resolution is dependent on the delivery optics and is limited by ocular aberrations. Through the combination of wavefront sensorless adaptive optics and the use of dual deformable transmissive optical elements, we present a compact lens-based OCT system at an imaging wavelength of 1060 nm for high resolution retinal imaging. We utilized a commercially available variable focal length lens to correct for a wide range of defocus commonly found in patient’s eyes, and a novel multi-actuator adaptive lens for aberration correction to achieve near diffraction limited imaging performance at the retina. With a parallel processing computational platform, high resolution cross-sectional and en face retinal image acquisition and display was performed in real time. In order to demonstrate the system functionality and clinical utility, we present images of the photoreceptor cone mosaic and other retinal layers acquired in vivo from research subjects.

  14. Fast super-resolution with affine motion using an adaptive Wiener filter and its application to airborne imaging.

    PubMed

    Hardie, Russell C; Barnard, Kenneth J; Ordonez, Raul

    2011-12-19

    Fast nonuniform interpolation based super-resolution (SR) has traditionally been limited to applications with translational interframe motion. This is in part because such methods are based on an underlying assumption that the warping and blurring components in the observation model commute. For translational motion this is the case, but it is not true in general. This presents a problem for applications such as airborne imaging where translation may be insufficient. Here we present a new Fourier domain analysis to show that, for many image systems, an affine warping model with limited zoom and shear approximately commutes with the point spread function when diffraction effects are modeled. Based on this important result, we present a new fast adaptive Wiener filter (AWF) SR algorithm for non-translational motion and study its performance with affine motion. The fast AWF SR method employs a new smart observation window that allows us to precompute all the needed filter weights for any type of motion without sacrificing much of the full performance of the AWF. We evaluate the proposed algorithm using simulated data and real infrared airborne imagery that contains a thermal resolution target allowing for objective resolution analysis.

  15. Diffraction-limited lucky imaging with a 12" commercial telescope

    NASA Astrophysics Data System (ADS)

    Baptista, Brian J.

    2014-08-01

    Here we demonstrate a novel lucky imaging camera which is designed to produce diffraction-limited imaging using small telescopes similar to ones used by many academic institutions for outreach and/or student training. We present a design that uses a Meade 12" SCT paired with an Andor iXon fast readout EMCCD. The PSF of the telescope is matched to the pixel size of the EMCCD by adding a simple, custom-fabricated, intervening optical system. We demonstrate performance of the system by observing both astronomical and terrestrial targets. The astronomical application requires simpler data reconstruction techniques as compared to the terrestrial case. We compare different lucky imaging registration and reconstruction algorithms for use with this imager for both astronomical and terrestrial targets. We also demonstrate how this type of instrument would be useful for both undergraduate and graduate student training. As an instructional aide, the instrument can provide a hands-on approach for teaching instrument design, standard data reduction techniques, lucky imaging data processing, and high resolution imaging concepts.

  16. Imaging System Model Crammed Into A 32K Microcomputer

    NASA Astrophysics Data System (ADS)

    Tyson, Robert K.

    1986-12-01

    An imaging system model, based upon linear systems theory, has been developed for a microcomputer with less than 32K of free random access memory (RAM). The model includes diffraction effects of the optics, aberrations in the optics, and atmospheric propagation transfer functions. Variables include pupil geometry, magnitude and character of the aberrations, and strength of atmospheric turbulence ("seeing"). Both coherent and incoherent image formation can be evaluated. The techniques employed for crowding the model into a very small computer will be discussed in detail. Simplifying assumptions for the diffraction and aberration phenomena will be shown along with practical considerations in modeling the optical system. Particular emphasis is placed on avoiding inaccuracies in modeling the pupil and the associated optical transfer function knowing limits on spatial frequency content and resolution. Memory and runtime constraints are analyzed stressing the efficient use of assembly language Fourier transform routines, disk input/output, and graphic displays. The compromises between computer time, limited RAM, and scientific accuracy will be given with techniques for balancing these parameters for individual needs.

  17. Near-Field Scanning Optical Microscopy and Raman Microscopy.

    NASA Astrophysics Data System (ADS)

    Harootunian, Alec Tate

    1987-09-01

    Both a one dimensional near-field scanning optical microscope and Raman microprobe were constructed. In near -field scanning optical microscopy (NSOM) a subwavelength aperture is scanned in the near-field of the object. Radiation transmitted through the aperture is collected to form an image as the aperture scans over the object. The resolution of an NSOM system is essentially wavelength independent and is limited by the diameter of the aperture used to scan the object. NSOM was developed in an effort to provide a nondestructive in situ high spatial resolution probe while still utilizing photons at optical wavelengths. The Raman microprobe constructed provided vibrational Raman information with spatial resolution equivalent that of a conventional diffraction limited microscope. Both transmission studies and near-field diffration studies of subwavelength apertures were performed. Diffraction theories for a small aperture in an infinitely thin conducting screen, a slit in a thick conducting screen, and an aperture in a black screen were examined. All three theories indicate collimation of radiation to the size to the size of the subwavelength aperture or slit in the near-field. Theoretical calculations and experimental results indicate that light transmitted through subwavelength apertures is readily detectable. Light of wavelength 4579 (ANGSTROM) was transmitted through apertures with diameters as small as 300 (ANGSTROM). These studies indicate the feasibility of constructing an NSOM system. One dimensional transmission and fluorescence NSOM systems were constructed. Apertures in the tips of metallized glass pipettes width inner diameters of less than 1000 (ANGSTROM) were used as a light source in the NSOM system. A tunneling current was used to maintain the aperture position in the near-field. Fluorescence NSOM was demonstrated for the first time. Microspectroscopic and Raman microscopic studies of turtle cone oil droplets were performed. Both the Raman vibrational frequencies and the Raman excitation data indicate that the carotenoids are unaggregated. The carotenoid astaxanthin was identified in the orange and red droplets by Raman microscopy. Future applications for both Raman microscopy and near-field microscopy were proposed. Four methods of near-field distance regulation were also examined. Finally, theoretical exposure curves for near-field lithography were calculated. Both the near-field lithographic results and the near field diffraction studies indicate essentially wavelength independent resolution. (Abstract shortened with permission of author.).

  18. Ultra-high Resolution Coherent X-ray Imaging of Nano-Materials

    NASA Astrophysics Data System (ADS)

    Shapiro, David

    A revolution is underway in the field of x-ray microscopy driven by the develop of experimental, theoretical and computational means of producing a complete description of coherent imaging systems from x-ray diffraction data. The methods being developed not only allow for full quantification and removal of all optical aberrations but also extension of the numerical aperture to the diffraction limit. One such method under intensive development is x-ray ptychography. This is a scanned probe method that reconstructs a scattering object and its illumination from coherent diffraction data. Within the first few years of development at the Advanced Light Source (ALS), Lawrence Berkeley National Laboratory, this method has already achieved the highest resolution x-ray images ever recorded in two, three and four dimensions. With the ability of x-rays to penetrate significantly more matter than electrons, their short wavelength and their sensitivity to chemical and magnetic states of matter, x-ray ptychography is set to revolutionize how we see the nano-scale world. In this presentation I will briefly describe the technical framework for how various methods work and will give a detailed account of a practical implementation at the ALS along with various scientific applications. The Advanced Light Source is supported by the Director, Office of Science, Office of Basic Energy Sciences, of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231.

  19. Imaging of sub-wavelength structures radiating coherently near microspheres

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Maslov, Alexey V., E-mail: avmaslov@yandex.ru; Astratov, Vasily N., E-mail: astratov@uncc.edu

    2016-02-01

    Using a two-dimensional model, we show that the optical images of a sub-wavelength object depend strongly on the excitation of its electromagnetic modes. There exist modes that enable the resolution of the object features smaller than the classical diffraction limit, in particular, due to the destructive interference. We propose to use such modes for super-resolution of resonant structures such as coupled cavities, metal dimers, or bowties. A dielectric microsphere in contact with the object forms its magnified image in a wide range of the virtual image plane positions. It is also suggested that the resonances may significantly affect the resolutionmore » quantification in recent experimental studies.« less

  20. In-line FINCH super resolution digital holographic fluorescence microscopy using a high efficiency transmission liquid crystal GRIN lens.

    PubMed

    Brooker, Gary; Siegel, Nisan; Rosen, Joseph; Hashimoto, Nobuyuki; Kurihara, Makoto; Tanabe, Ayano

    2013-12-15

    We report a new optical arrangement that creates high-efficiency, high-quality Fresnel incoherent correlation holography (FINCH) holograms using polarization sensitive transmission liquid crystal gradient index (TLCGRIN) diffractive lenses. In contrast, current universal practice in the field employs a reflective spatial light modulator (SLM) to separate sample and reference beams. Polarization sensitive TLCGRIN lenses enable a straight optical path, have >90% transmission efficiency, are not pixilated, and are free of many limitations of reflective SLM devices. For each sample point, two spherical beams created by a glass lens in combination with a polarization sensitive TLCGRIN lens interfere and create a hologram and resultant super resolution image.

  1. Snorkelling between the stars: submarine methods for astronomical observations.

    NASA Astrophysics Data System (ADS)

    Velasco, S.; Quevedo, E.; Font, J.; Oscoz, A.; López, R. L.; Puga, M.; Rebolo, R.; Hernáandez Brito, J.; Llinas, O.; Marrero Callico, G.; Sarmiento, R.

    2017-03-01

    Trying to reach diffraction-limited astronomical observations from ground-based telescopes is very challenging due to the atmospheric effects contributing to a general blurring of the images. However, astronomy is not the only science facing turbulence problems; obtaining quality images of the undersea world is as ambitious as it is on the sky. One of the solutions contemplated to reach high-resolution images is the use of multiple frames of the same target, known as fusion super-resolution (Quevedo et al. 2015), which is the principle for Lucky Imaging (Velasco et al. 2016). Here we present the successful result of joining efforts between the undersea and the astronomical research done at the Canary Islands.

  2. Neptune and Titan Observed with Keck Telescope Adaptive Optics

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Max, C.E.; Macintosh, B.A.; Gibbard, S.

    2000-05-05

    The authors report on observations taken during engineering science validation time using the new adaptive optics system at the 10-m Keck II Telescope. They observe Neptune and Titan at near-infrared wavelengths. These objects are ideal for adaptive optics imaging because they are bright and small, yet have many diffraction-limited resolution elements across their disks. In addition Neptune and Titan have prominent physical features, some of which change markedly with time. They have observed infrared-bright storms on Neptune, and very low-albedo surface regions on Titan, Saturn's largest moon, Spatial resolution on Neptune and Titan was 0.05-0.06 and 0.04-0.05 arc sec, respectively.

  3. Fabrication of high-resolution x-ray diffractive optics at King's College London

    NASA Astrophysics Data System (ADS)

    Charalambous, Pambos S.; Anastasi, Peter A. F.; Burge, Ronald E.; Popova, Katia

    1995-09-01

    The fabrication of high resolution x-ray diffractive optics, and Fresnel zone plates (ZPs) in particular, is a very demanding multifaceted technological task. The commissioning of more (and brighter) synchrotron radiation sources, has increased the number of x-ray imaging beam lines world wide. The availability of cheaper and more effective laboratory x-ray sources, has further increased the number of laboratories involved in x-ray imaging. The result is an ever increasing demand for x-ray optics with a very wide range of specifications, reflecting the particular type of x-ray imaging performed at different laboratories. We have been involved in all aspects of high resolution nanofabrication for a number of years, and we have explored many different methods of lithography, which, although unorthodox, open up possibilities, and increase our flexibility for the fabrication of different diffractive optical elements, as well as other types of nanostructures. The availability of brighter x-ray sources, means that the diffraction efficiency of the ZPs is becoming of secondary importance, a trend which will continue in the future. Resolution, however, is important and will always remain so. Resolution is directly related to the accuracy af pattern generation, as well as the ability to draw fine lines. This is the area towards which we have directed most of our efforts so far.

  4. Adaptive Optics Optical Coherence Tomography in Glaucoma

    PubMed Central

    Dong, Zachary M.; Wollstein, Gadi; Wang, Bo; Schuman, Joel S.

    2016-01-01

    Since the introduction of commercial optical coherence tomography (OCT) systems, the ophthalmic imaging modality has rapidly expanded and it has since changed the paradigm of visualization of the retina and revolutionized the management and diagnosis of neuro-retinal diseases, including glaucoma. OCT remains a dynamic and evolving imaging modality, growing from time-domain OCT to the improved spectral-domain OCT, adapting novel image analysis and processing methods, and onto the newer swept-source OCT and the implementation of adaptive optics (AO) into OCT. The incorporation of AO into ophthalmic imaging modalities has enhanced OCT by improving image resolution and quality, particularly in the posterior segment of the eye. Although OCT previously captured in-vivo cross-sectional images with unparalleled high resolution in the axial direction, monochromatic aberrations of the eye limit transverse or lateral resolution to about 15-20 μm and reduce overall image quality. In pairing AO technology with OCT, it is now possible to obtain diffraction-limited resolution images of the optic nerve head and retina in three-dimensions, increasing resolution down to a theoretical 3 μm3. It is now possible to visualize discrete structures within the posterior eye, such as photoreceptors, retinal nerve fiber layer bundles, the lamina cribrosa, and other structures relevant to glaucoma. Despite its limitations and barriers to widespread commercialization, the expanding role of AO in OCT is propelling this technology into clinical trials and onto becoming an invaluable modality in the clinician's arsenal. PMID:27916682

  5. A versatile genome-scale PCR-based pipeline for high-definition DNA FISH.

    PubMed

    Bienko, Magda; Crosetto, Nicola; Teytelman, Leonid; Klemm, Sandy; Itzkovitz, Shalev; van Oudenaarden, Alexander

    2013-02-01

    We developed a cost-effective genome-scale PCR-based method for high-definition DNA FISH (HD-FISH). We visualized gene loci with diffraction-limited resolution, chromosomes as spot clusters and single genes together with transcripts by combining HD-FISH with single-molecule RNA FISH. We provide a database of over 4.3 million primer pairs targeting the human and mouse genomes that is readily usable for rapid and flexible generation of probes.

  6. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hosseinizadeh, Ahmad; Mashayekhi, Ghoncheh; Copperman, Jeremy

    Using a manifold-based analysis of experimental diffraction snapshots from an X-ray free electron laser, we determine the three-dimensional structure and conformational landscape of the PR772 virus to a detector-limited resolution of 9 nm. Our results indicate that a single conformational coordinate controls reorganization of the genome, growth of a tubular structure from a portal vertex and release of the genome. Furthermore, these results demonstrate that single-particle X-ray scattering has the potential to shed light on key biological processes.

  7. Multiplexed aberration measurement for deep tissue imaging in vivo

    PubMed Central

    Wang, Chen; Liu, Rui; Milkie, Daniel E.; Sun, Wenzhi; Tan, Zhongchao; Kerlin, Aaron; Chen, Tsai-Wen; Kim, Douglas S.; Ji, Na

    2014-01-01

    We describe a multiplexed aberration measurement method that modulates the intensity or phase of light rays at multiple pupil segments in parallel to determine their phase gradients. Applicable to fluorescent-protein-labeled structures of arbitrary complexity, it allows us to obtain diffraction-limited resolution in various samples in vivo. For the strongly scattering mouse brain, a single aberration correction improves structural and functional imaging of fine neuronal processes over a large imaging volume. PMID:25128976

  8. Wavefront correction and high-resolution in vivo OCT imaging with an objective integrated multi-actuator adaptive lens.

    PubMed

    Bonora, Stefano; Jian, Yifan; Zhang, Pengfei; Zam, Azhar; Pugh, Edward N; Zawadzki, Robert J; Sarunic, Marinko V

    2015-08-24

    Adaptive optics is rapidly transforming microscopy and high-resolution ophthalmic imaging. The adaptive elements commonly used to control optical wavefronts are liquid crystal spatial light modulators and deformable mirrors. We introduce a novel Multi-actuator Adaptive Lens that can correct aberrations to high order, and which has the potential to increase the spread of adaptive optics to many new applications by simplifying its integration with existing systems. Our method combines an adaptive lens with an imaged-based optimization control that allows the correction of images to the diffraction limit, and provides a reduction of hardware complexity with respect to existing state-of-the-art adaptive optics systems. The Multi-actuator Adaptive Lens design that we present can correct wavefront aberrations up to the 4th order of the Zernike polynomial characterization. The performance of the Multi-actuator Adaptive Lens is demonstrated in a wide field microscope, using a Shack-Hartmann wavefront sensor for closed loop control. The Multi-actuator Adaptive Lens and image-based wavefront-sensorless control were also integrated into the objective of a Fourier Domain Optical Coherence Tomography system for in vivo imaging of mouse retinal structures. The experimental results demonstrate that the insertion of the Multi-actuator Objective Lens can generate arbitrary wavefronts to correct aberrations down to the diffraction limit, and can be easily integrated into optical systems to improve the quality of aberrated images.

  9. High spatial resolution X-ray and gamma ray imaging system using diffraction crystals

    DOEpatents

    Smither, Robert K [Hinsdale, IL

    2011-05-17

    A method and a device for high spatial resolution imaging of a plurality of sources of x-ray and gamma-ray radiation are provided. The device comprises a plurality of arrays, with each array comprising a plurality of elements comprising a first collimator, a diffracting crystal, a second collimator, and a detector.

  10. Incoherent Diffractive Imaging via Intensity Correlations of Hard X Rays

    NASA Astrophysics Data System (ADS)

    Classen, Anton; Ayyer, Kartik; Chapman, Henry N.; Röhlsberger, Ralf; von Zanthier, Joachim

    2017-08-01

    Established x-ray diffraction methods allow for high-resolution structure determination of crystals, crystallized protein structures, or even single molecules. While these techniques rely on coherent scattering, incoherent processes like fluorescence emission—often the predominant scattering mechanism—are generally considered detrimental for imaging applications. Here, we show that intensity correlations of incoherently scattered x-ray radiation can be used to image the full 3D arrangement of the scattering atoms with significantly higher resolution compared to conventional coherent diffraction imaging and crystallography, including additional three-dimensional information in Fourier space for a single sample orientation. We present a number of properties of incoherent diffractive imaging that are conceptually superior to those of coherent methods.

  11. Computer synthesis of high resolution electron micrographs

    NASA Technical Reports Server (NTRS)

    Nathan, R.

    1976-01-01

    Specimen damage, spherical aberration, low contrast and noisy sensors combine to prevent direct atomic viewing in a conventional electron microscope. The paper describes two methods for obtaining ultra-high resolution in biological specimens under the electron microscope. The first method assumes the physical limits of the electron objective lens and uses a series of dark field images of biological crystals to obtain direct information on the phases of the Fourier diffraction maxima; this information is used in an appropriate computer to synthesize a large aperture lens for a 1-A resolution. The second method assumes there is sufficient amplitude scatter from images recorded in focus which can be utilized with a sensitive densitometer and computer contrast stretching to yield fine structure image details. Cancer virus characterization is discussed as an illustrative example. Numerous photographs supplement the text.

  12. From single-molecule spectroscopy to super-resolution imaging of the neuron: a review

    PubMed Central

    Laine, Romain F; Kaminski Schierle, Gabriele S; van de Linde, Sebastian; Kaminski, Clemens F

    2016-01-01

    Abstract For more than 20 years, single-molecule spectroscopy has been providing invaluable insights into nature at the molecular level. The field has received a powerful boost with the development of the technique into super-resolution imaging methods, ca. 10 years ago, which overcome the limitations imposed by optical diffraction. Today, single molecule super-resolution imaging is routinely used in the study of macromolecular function and structure in the cell. Concomitantly, computational methods have been developed that provide information on numbers and positions of molecules at the nanometer-scale. In this overview, we outline the technical developments that have led to the emergence of localization microscopy techniques from single-molecule spectroscopy. We then provide a comprehensive review on the application of the technique in the field of neuroscience research. PMID:28809165

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

    PubMed Central

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

    2016-01-01

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

  14. Sub-atomic resolution X-ray crystallography and neutron crystallography: promise, challenges and potential.

    PubMed

    Blakeley, Matthew P; Hasnain, Samar S; Antonyuk, Svetlana V

    2015-07-01

    The International Year of Crystallography saw the number of macromolecular structures deposited in the Protein Data Bank cross the 100000 mark, with more than 90000 of these provided by X-ray crystallography. The number of X-ray structures determined to sub-atomic resolution (i.e. ≤1 Å) has passed 600 and this is likely to continue to grow rapidly with diffraction-limited synchrotron radiation sources such as MAX-IV (Sweden) and Sirius (Brazil) under construction. A dozen X-ray structures have been deposited to ultra-high resolution (i.e. ≤0.7 Å), for which precise electron density can be exploited to obtain charge density and provide information on the bonding character of catalytic or electron transfer sites. Although the development of neutron macromolecular crystallography over the years has been far less pronounced, and its application much less widespread, the availability of new and improved instrumentation, combined with dedicated deuteration facilities, are beginning to transform the field. Of the 83 macromolecular structures deposited with neutron diffraction data, more than half (49/83, 59%) were released since 2010. Sub-mm(3) crystals are now regularly being used for data collection, structures have been determined to atomic resolution for a few small proteins, and much larger unit-cell systems (cell edges >100 Å) are being successfully studied. While some details relating to H-atom positions are tractable with X-ray crystallography at sub-atomic resolution, the mobility of certain H atoms precludes them from being located. In addition, highly polarized H atoms and protons (H(+)) remain invisible with X-rays. Moreover, the majority of X-ray structures are determined from cryo-cooled crystals at 100 K, and, although radiation damage can be strongly controlled, especially since the advent of shutterless fast detectors, and by using limited doses and crystal translation at micro-focus beams, radiation damage can still take place. Neutron crystallography therefore remains the only approach where diffraction data can be collected at room temperature without radiation damage issues and the only approach to locate mobile or highly polarized H atoms and protons. Here a review of the current status of sub-atomic X-ray and neutron macromolecular crystallography is given and future prospects for combined approaches are outlined. New results from two metalloproteins, copper nitrite reductase and cytochrome c', are also included, which illustrate the type of information that can be obtained from sub-atomic-resolution (∼0.8 Å) X-ray structures, while also highlighting the need for complementary neutron studies that can provide details of H atoms not provided by X-ray crystallography.

  15. Towards the development of a hybrid-integrated chip interferometer for online surface profile measurements

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kumar, P.; Martin, H.; Jiang, X.

    Non-destructive testing and online measurement of surface features are pressing demands in manufacturing. Thus optical techniques are gaining importance for characterization of complex engineering surfaces. Harnessing integrated optics for miniaturization of interferometry systems onto a silicon wafer and incorporating a compact optical probe would enable the development of a handheld sensor for embedded metrology applications. In this work, we present the progress in the development of a hybrid photonics based metrology sensor device for online surface profile measurements. The measurement principle along with test and measurement results of individual components has been presented. For non-contact measurement, a spectrally encoded lateralmore » scanning probe based on the laser scanning microscopy has been developed to provide fast measurement with lateral resolution limited to the diffraction limit. The probe demonstrates a lateral resolution of ∼3.6 μm while high axial resolution (sub-nanometre) is inherently achieved by interferometry. Further the performance of the hybrid tuneable laser and the scanning probe was evaluated by measuring a standard step height sample of 100 nm.« less

  16. Microlens performance limits in sub-2mum pixel CMOS image sensors.

    PubMed

    Huo, Yijie; Fesenmaier, Christian C; Catrysse, Peter B

    2010-03-15

    CMOS image sensors with smaller pixels are expected to enable digital imaging systems with better resolution. When pixel size scales below 2 mum, however, diffraction affects the optical performance of the pixel and its microlens, in particular. We present a first-principles electromagnetic analysis of microlens behavior during the lateral scaling of CMOS image sensor pixels. We establish for a three-metal-layer pixel that diffraction prevents the microlens from acting as a focusing element when pixels become smaller than 1.4 microm. This severely degrades performance for on and off-axis pixels in red, green and blue color channels. We predict that one-metal-layer or backside-illuminated pixels are required to extend the functionality of microlenses beyond the 1.4 microm pixel node.

  17. Coherent diffraction imaging analysis of shape-controlled nanoparticles with focused hard X-ray free-electron laser pulses.

    PubMed

    Takahashi, Yukio; Suzuki, Akihiro; Zettsu, Nobuyuki; Oroguchi, Tomotaka; Takayama, Yuki; Sekiguchi, Yuki; Kobayashi, Amane; Yamamoto, Masaki; Nakasako, Masayoshi

    2013-01-01

    We report the first demonstration of the coherent diffraction imaging analysis of nanoparticles using focused hard X-ray free-electron laser pulses, allowing us to analyze the size distribution of particles as well as the electron density projection of individual particles. We measured 1000 single-shot coherent X-ray diffraction patterns of shape-controlled Ag nanocubes and Au/Ag nanoboxes and estimated the edge length from the speckle size of the coherent diffraction patterns. We then reconstructed the two-dimensional electron density projection with sub-10 nm resolution from selected coherent diffraction patterns. This method enables the simultaneous analysis of the size distribution of synthesized nanoparticles and the structures of particles at nanoscale resolution to address correlations between individual structures of components and the statistical properties in heterogeneous systems such as nanoparticles and cells.

  18. Spiders in Lyot Coronagraphs

    NASA Astrophysics Data System (ADS)

    Sivaramakrishnan, Anand; Lloyd, James P.

    2005-11-01

    In principle, suppression of on-axis stellar light by a coronagraph is easier on an unobscured aperture telescope than on one with an obscured aperture. Recent designs such as the apodized pupil Lyot coronagraph, the ``band-limited'' Lyot coronagraph, and several variants of phase-mask coronagraphs work best on unobscured circular aperture telescopes. These designs were developed to enable the discovery and characterization of nearby Jovian or even terrestrial exoplanets. All of today's major space-based and adaptive optics-equipped ground-based telescopes are obscured-aperture systems with a secondary mirror held in place by secondary support ``spider'' vanes. The presence of a secondary obscuration can be dealt with by ingenious coronagraph designs, but the spider vanes themselves cause diffracted light, which can hamper the search for Jovian exoplanets around nearby stars. We look at the problem of suppressing spider vane diffraction in Lyot coronagraphs, including apodized pupil and band-limited designs. We show how spider vane diffraction can be reduced drastically and in fact contained in the final coronagraphic image, within one resolution element of the geometric image of the focal plane mask's occulting spot. This makes adaptive optics coronagraphic searches for exojupiters possible with the next generation of adaptive optics systems being developed for 8-10 m class telescopes such as Gemini and the Very Large Telescopes.

  19. Reaching the Diffraction Limit - Differential Speckle and Wide-Field Imaging for the WIYN Telescope

    NASA Technical Reports Server (NTRS)

    Scott, Nic J.; Howell, Steve; Horch, Elliott

    2016-01-01

    Speckle imaging allows telescopes to achieve diffraction limited imaging performance. The technique requires cameras capable of reading out frames at a very fast rate, effectively 'freezing out' atmospheric seeing. The resulting speckles can be correlated and images reconstructed that are at the diffraction limit of the telescope. These new instruments are based on the successful performance and design of the Differential Speckle Survey Instrument (DSSI).The instruments are being built for the Gemini-N and WIYN telescopes and will be made available to the community via the peer review proposal process. We envision their primary use to be validation and characterization of exoplanet targets from the NASA, K2 and TESS missions and RV discovered exoplanets. Such targets will provide excellent follow-up candidates for both the WIYN and Gemini telescopes. We expect similar data quality in speckle imaging mode with the new instruments. Additionally, both cameras will have a wide-field mode and standard SDSS filters. They will be highly versatile instruments and it is that likely many other science programs will request time on the cameras. The limiting magnitude for speckle observations will remain around 13-14th at WIYN and 16-17th at Gemini, while wide-field, normal CCD imaging operation should be able to go to much fainter, providing usual CCD imaging and photometric capabilities. The instruments will also have high utility as scoring cameras for telescope engineering purposes, or other applications where high time resolution is needed. Instrument support will be provided, including a software pipeline that takes raw speckle data to fully reconstructed images.

  20. Structural molecular biology: Recent results from neutron diffraction

    NASA Astrophysics Data System (ADS)

    Timmins, Peter A.

    1995-02-01

    Neutron diffraction is of importance in structural biology at several different levels of resolution. In most cases the unique possibility arising from deuterium labelling or contrast variation is of fundamental importance in providing information complementary to that which can be obtained from X-ray diffraction. At high resolution, neutron crystallography of proteins allows the location of hydrogen atoms in the molecule or of the hydration water, both of which may be central to biological activity. A major difficulty in this field has been the poor signal-to-noise ratio of the data arising not only from relatively low beam intensities and small crystals but, most importantly from the incoherent background due to hydrogen atoms in the sample. Modern methods of molecular biology now offer ways of producing fully deuterated proteins by cloning in bacteria grown on fully deuterated media. At a slightly lower resolution, there are a number of systems which may be ordered in one or two dimensions. This is the case in the purple membrane where neutron diffraction with deuterium labelling has complemented high resolution electron diffraction. Finally there is a class of very large macromolecular systems which can be crystallised and have been studied by X-ray diffraction but in which part of the structure is locally disordered and usually has insufficient contrast to be seen with X-rays. In this case the use of H 2O/D 2O contrast variation allows these components to be located. Examples of this are the nucleic acid in virus structures and detergent bound to membrane proteins.

  1. Experiences with making diffraction image data available: what metadata do we need to archive?

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kroon-Batenburg, Loes M. J., E-mail: l.m.j.kroon-batenburg@uu.nl; Helliwell, John R.; Utrecht University, Padualaan 8, 3584 CH Utrecht

    A local raw ‘diffraction data images’ archive was made available and some data sets were retrieved and reprocessed, which led to analysis of the anomalous difference densities of two partially occupied Cl atoms in cisplatin as well as a re-evaluation of the resolution cutoff in these diffraction data. General questions on storing raw data are discussed. It is also demonstrated that often one needs unambiguous prior knowledge to read the (binary) detector format and the setup of goniometer geometries. Recently, the IUCr (International Union of Crystallography) initiated the formation of a Diffraction Data Deposition Working Group with the aim ofmore » developing standards for the representation of raw diffraction data associated with the publication of structural papers. Archiving of raw data serves several goals: to improve the record of science, to verify the reproducibility and to allow detailed checks of scientific data, safeguarding against fraud and to allow reanalysis with future improved techniques. A means of studying this issue is to submit exemplar publications with associated raw data and metadata. In a recent study of the binding of cisplatin and carboplatin to histidine in lysozyme crystals under several conditions, the possible effects of the equipment and X-ray diffraction data-processing software on the occupancies and B factors of the bound Pt compounds were compared. Initially, 35.3 GB of data were transferred from Manchester to Utrecht to be processed with EVAL. A detailed description and discussion of the availability of metadata was published in a paper that was linked to a local raw data archive at Utrecht University and also mirrored at the TARDIS raw diffraction data archive in Australia. By making these raw diffraction data sets available with the article, it is possible for the diffraction community to make their own evaluation. This led to one of the authors of XDS (K. Diederichs) to re-integrate the data from crystals that supposedly solely contained bound carboplatin, resulting in the analysis of partially occupied chlorine anomalous electron densities near the Pt-binding sites and the use of several criteria to more carefully assess the diffraction resolution limit. General arguments for archiving raw data, the possibilities of doing so and the requirement of resources are discussed. The problems associated with a partially unknown experimental setup, which preferably should be available as metadata, is discussed. Current thoughts on data compression are summarized, which could be a solution especially for pixel-device data sets with fine slicing that may otherwise present an unmanageable amount of data.« less

  2. Purification, crystallization and preliminary X-ray analysis of a thermostable glycoside hydrolase family 43 β-xylosidase from Geobacillus thermoleovorans IT-08

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rohman, Ali; Oosterwijk, Niels van; Kralj, Slavko

    2007-11-01

    The β-xylosidase was crystallized using PEG 6000 as precipitant. 5% PEG 6000 yielded bipyramid-shaped tetragonal crystals diffracting to 1.55 Å resolution, and 13% PEG 6000 gave rectangular monoclinic crystals diffracting to 1.80 Å resolution. The main enzymes involved in xylan-backbone hydrolysis are endo-1,4-β-xylanase and β-xylosidase. β-Xylosidase converts the xylo-oligosaccharides produced by endo-1,4-β-xylanase into xylose monomers. The β-xylosidase from the thermophilic Geobacillus thermoleovorans IT-08, a member of glycoside hydrolase family 43, was crystallized at room temperature using the hanging-drop vapour-diffusion method. Two crystal forms were observed. Bipyramid-shaped crystals belonging to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = bmore » = 62.53, c = 277.4 Å diffracted to 1.55 Å resolution. The rectangular crystals belonged to space group P2{sub 1}, with unit-cell parameters a = 57.94, b = 142.1, c = 153.9 Å, β = 90.5°, and diffracted to 1.80 Å resolution.« less

  3. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tregillis, Ian Lee

    This document examines the performance of a generic flat-mirror multimonochromatic imager (MMI), with special emphasis on existing instruments at NIF and Omega. We begin by deriving the standard equation for the mean number of photons detected per resolution element. The pinhole energy bandwidth is a contributing factor; this is dominated by the finite size of the source and may be considerable. The most common method for estimating the spatial resolution of such a system (quadrature addition) is, technically, mathematically invalid for this case. However, under the proper circumstances it may produce good estimates compared to a rigorous calculation based onmore » the convolution of point-spread functions. Diffraction is an important contribution to the spatial resolution. Common approximations based on Fraunhofer (farfield) diffraction may be inappropriate and misleading, as the instrument may reside in multiple regimes depending upon its configuration or the energy of interest. It is crucial to identify the correct diffraction regime; Fraunhofer and Fresnel (near-field) diffraction profiles are substantially different, the latter being considerably wider. Finally, we combine the photonics and resolution analyses to derive an expression for the minimum signal level such that the resulting images are not dominated by photon statistics. This analysis is consistent with observed performance of the NIF MMI.« less

  4. Kernel-Phase Interferometry for Super-Resolution Detection of Faint Companions

    NASA Astrophysics Data System (ADS)

    Factor, Samuel M.; Kraus, Adam L.

    2017-06-01

    Direct detection of close in companions (exoplanets or binary systems) is notoriously difficult. While coronagraphs and point spread function (PSF) subtraction can be used to reduce contrast and dig out signals of companions under the PSF, there are still significant limitations in separation and contrast near λ/D. Non-redundant aperture masking (NRM) interferometry can be used to detect companions well inside the PSF of a diffraction limited image, though the mask discards ˜ 95% of the light gathered by the telescope and thus the technique is severely flux limited. Kernel-phase analysis applies interferometric techniques similar to NRM to a diffraction limited image utilizing the full aperture. Instead of non-redundant closure-phases, kernel-phases are constructed from a grid of points on the full aperture, simulating a redundant interferometer. I have developed a new, easy to use, faint companion detection pipeline which analyzes kernel-phases utilizing Bayesian model comparison. I demonstrate this pipeline on archival images from HST/NICMOS, searching for new companions in order to constrain binary formation models at separations inaccessible to previous techniques. Using this method, it is possible to detect a companion well within the classical λ/D Rayleigh diffraction limit using a fraction of the telescope time as NRM. Since the James Webb Space Telescope (JWST) will be able to perform NRM observations, further development and characterization of kernel-phase analysis will allow efficient use of highly competitive JWST telescope time. As no mask is needed, this technique can easily be applied to archival data and even target acquisition images (e.g. from JWST), making the detection of close in companions cheap and simple as no additional observations are needed.

  5. Coherent x-ray diffraction

    NASA Astrophysics Data System (ADS)

    Pitney, John Allen

    Conventional x-ray diffraction has historically been done under conditions such that the measured signal consists of an incoherent addition of scattering which is coherent only on a length scale determined by the properties of the beam. The result of the incoherent summation is a statistical averaging over the whole illuminated volume of the sample, which yields certain kinds of information with a high degree of precision and has been key to the success of x-ray diffraction in a variety of applications. Coherent x-ray scattering techniques, such as coherent x-ray diffraction (CXD) and x-ray intensity fluctuation spectroscopy (XIFS), attempt to reduce or eliminate any incoherent averaging so that specific, local structures couple to the measurement without being averaged out. In the case of XIFS, the result is analogous to dynamical light scattering, but with sensitivity to length scales less than 200 nm and time scales from 10-3 s to 103 s. When combined with phase retrieval, CXD represents an imaging technique with the penetration, in situ capabilities, and contrast mechanisms associated with x-rays and with a spatial resolution ultimately limited by the x-ray wavelength. In practice, however, the spatial resolution of CXD imaging is limited by exposure to about 100 A. This thesis describes CXD measurements of the binary alloy Cu3Au and the adaptation of phase retrieval methods for the reconstruction of real-space images of Cu3Au antiphase domains. The theoretical foundations of CXD are described in Chapter 1 as derived from the kinematical formulation for x-ray diffraction and from the temporal and spatial coherence of radiation. The antiphase domain structure of Cu 3Au is described, along with the associated reciprocal-space structure which is measured by CXD. CXD measurements place relatively stringent requirements on the coherence properties of the beam and on the detection mechanism of the experiment; these requirements and the means by which they have been met are delineated in Chapter 2. The results and interpretation of a set of Cu 3Au measurements are presented in Chapter 3. Chapter 4 describes the Gerchberg-Saxton and the hybrid input-output (HIO) algorithms for phase retrieval and shows the results of image reconstruction tests with simulated Cu 3Au CXD, including the effect of oversampling in reciprocal space.

  6. Imaging single cells in a beam of live cyanobacteria with an X-ray laser.

    PubMed

    van der Schot, Gijs; Svenda, Martin; Maia, Filipe R N C; Hantke, Max; DePonte, Daniel P; Seibert, M Marvin; Aquila, Andrew; Schulz, Joachim; Kirian, Richard; Liang, Mengning; Stellato, Francesco; Iwan, Bianca; Andreasson, Jakob; Timneanu, Nicusor; Westphal, Daniel; Almeida, F Nunes; Odic, Dusko; Hasse, Dirk; Carlsson, Gunilla H; Larsson, Daniel S D; Barty, Anton; Martin, Andrew V; Schorb, Sebastian; Bostedt, Christoph; Bozek, John D; Rolles, Daniel; Rudenko, Artem; Epp, Sascha; Foucar, Lutz; Rudek, Benedikt; Hartmann, Robert; Kimmel, Nils; Holl, Peter; Englert, Lars; Duane Loh, Ne-Te; Chapman, Henry N; Andersson, Inger; Hajdu, Janos; Ekeberg, Tomas

    2015-02-11

    There exists a conspicuous gap of knowledge about the organization of life at mesoscopic levels. Ultra-fast coherent diffractive imaging with X-ray free-electron lasers can probe structures at the relevant length scales and may reach sub-nanometer resolution on micron-sized living cells. Here we show that we can introduce a beam of aerosolised cyanobacteria into the focus of the Linac Coherent Light Source and record diffraction patterns from individual living cells at very low noise levels and at high hit ratios. We obtain two-dimensional projection images directly from the diffraction patterns, and present the results as synthetic X-ray Nomarski images calculated from the complex-valued reconstructions. We further demonstrate that it is possible to record diffraction data to nanometer resolution on live cells with X-ray lasers. Extension to sub-nanometer resolution is within reach, although improvements in pulse parameters and X-ray area detectors will be necessary to unlock this potential.

  7. High-angular-resolution NIR astronomy with large arrays (SHARP I and SHARP II)

    NASA Astrophysics Data System (ADS)

    Hofmann, Reiner; Brandl, Bernhard; Eckart, Andreas; Eisenhauer, Frank; Tacconi-Garman, Lowell E.

    1995-06-01

    SHARP I and SHARP II are near infrared cameras for high-angular-resolution imaging. Both cameras are built around a 256 X 256 pixel NICMOS 3 HgCdTe array from Rockwell which is sensitive in the 1 - 2.5 micrometers range. With a 0.05'/pixel scale, they can produce diffraction limited K-band images at 4-m-class telescopes. For a 256 X 256 array, this pixel scale results in a field of view of 12.8' X 12.8' which is well suited for the observation of galactic and extragalactic near-infrared sources. Photometric and low resolution spectroscopic capabilities are added by photometric band filters (J, H, K), narrow band filters ((lambda) /(Delta) (lambda) approximately equals 100) for selected spectral lines, and a CVF ((lambda) /(Delta) (lambda) approximately equals 70). A cold shutter permits short exposure times down to about 10 ms. The data acquisition electronics permanently accepts the maximum frame rate of 8 Hz which is defined by the detector time constants (data rate 1 Mbyte/s). SHARP I has been especially designed for speckle observations at ESO's 3.5 m New Technology Telescope and is in operation since 1991. SHARP II is used at ESO's 3.6 m telescope together with the adaptive optics system COME-ON + since 1993. A new version of SHARP II is presently under test, which incorporates exchangeable camera optics for observations with scales of 0.035, 0.05, and 0.1'/pixel. The first scale extends diffraction limited observations down to the J-band, while the last one provides a larger field of view. To demonstrate the power of the cameras, images of the galactic center obtained with SHARP I, and images of the R136 region in 30 Doradus observed with SHARP II are presented.

  8. Revealing the Effects of Nanoscale Membrane Curvature on Lipid Mobility

    PubMed Central

    Kabbani, Abir Maarouf; Woodward, Xinxin

    2017-01-01

    Recent advances in nanoengineering and super-resolution microscopy have enabled new capabilities for creating and observing membrane curvature. However, the effects of curvature on single-lipid diffusion have yet to be revealed. The simulations presented here describe the capabilities of varying experimental methods for revealing the effects of nanoscale curvature on single-molecule mobility. Traditionally, lipid mobility is revealed through fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), and single particle tracking (SPT). However, these techniques vary greatly in their ability to detect the effects of nanoscale curvature on lipid behavior. Traditionally, FRAP and FCS depend on diffraction-limited illumination and detection. A simulation of FRAP shows minimal effects on lipids diffusion due to a 50 nm radius membrane bud. Throughout the stages of the budding process, FRAP detected minimal changes in lipid recovery time due to the curvature versus flat membrane. Simulated FCS demonstrated small effects due to a 50 nm radius membrane bud that was more apparent with curvature-dependent lipid mobility changes. However, SPT achieves a sub-diffraction-limited resolution of membrane budding and lipid mobility through the identification of the single-lipid positions with ≤15 nm spatial and ≤20 ms temporal resolution. By mapping the single-lipid step lengths to locations on the membrane, the effects of membrane topography and curvature could be correlated to the effective membrane viscosity. Single-fluorophore localization techniques, such SPT, can detect membrane curvature and its effects on lipid behavior. These simulations and discussion provide a guideline for optimizing the experimental procedures in revealing the effects of curvature on lipid mobility and effective local membrane viscosity. PMID:29057801

  9. High quality single shot ultrafast MeV electron diffraction from a photocathode radio-frequency gun

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fu, Feichao; Liu, Shengguang; Zhu, Pengfei

    2014-08-15

    A compact ultrafast electron diffractometer, consisting of an s-band 1.6 cell photocathode radio-frequency gun, a multi-function changeable sample chamber, and a sensitive relativistic electron detector, was built at Shanghai Jiao Tong University. High-quality single-shot transmission electron diffraction patterns have been recorded by scattering 2.5 MeV electrons off single crystalline gold and polycrystalline aluminum samples. The high quality diffraction pattern indicates an excellent spatial resolution, with the ratio of the diffraction ring radius over the ring rms width beyond 10. The electron pulse width is estimated to be about 300 fs. The high temporal and spatial resolution may open new opportunities inmore » various areas of sciences.« less

  10. High quality single shot ultrafast MeV electron diffraction from a photocathode radio-frequency gun.

    PubMed

    Fu, Feichao; Liu, Shengguang; Zhu, Pengfei; Xiang, Dao; Zhang, Jie; Cao, Jianming

    2014-08-01

    A compact ultrafast electron diffractometer, consisting of an s-band 1.6 cell photocathode radio-frequency gun, a multi-function changeable sample chamber, and a sensitive relativistic electron detector, was built at Shanghai Jiao Tong University. High-quality single-shot transmission electron diffraction patterns have been recorded by scattering 2.5 MeV electrons off single crystalline gold and polycrystalline aluminum samples. The high quality diffraction pattern indicates an excellent spatial resolution, with the ratio of the diffraction ring radius over the ring rms width beyond 10. The electron pulse width is estimated to be about 300 fs. The high temporal and spatial resolution may open new opportunities in various areas of sciences.

  11. Accurate macromolecular structures using minimal measurements from X-ray free-electron lasers

    PubMed Central

    Hattne, Johan; Echols, Nathaniel; Tran, Rosalie; Kern, Jan; Gildea, Richard J.; Brewster, Aaron S.; Alonso-Mori, Roberto; Glöckner, Carina; Hellmich, Julia; Laksmono, Hartawan; Sierra, Raymond G.; Lassalle-Kaiser, Benedikt; Lampe, Alyssa; Han, Guangye; Gul, Sheraz; DiFiore, Dörte; Milathianaki, Despina; Fry, Alan R.; Miahnahri, Alan; White, William E.; Schafer, Donald W.; Seibert, M. Marvin; Koglin, Jason E.; Sokaras, Dimosthenis; Weng, Tsu-Chien; Sellberg, Jonas; Latimer, Matthew J.; Glatzel, Pieter; Zwart, Petrus H.; Grosse-Kunstleve, Ralf W.; Bogan, Michael J.; Messerschmidt, Marc; Williams, Garth J.; Boutet, Sébastien; Messinger, Johannes; Zouni, Athina; Yano, Junko; Bergmann, Uwe; Yachandra, Vittal K.; Adams, Paul D.; Sauter, Nicholas K.

    2014-01-01

    X-ray free-electron laser (XFEL) sources enable the use of crystallography to solve three-dimensional macromolecular structures under native conditions and free from radiation damage. Results to date, however, have been limited by the challenge of deriving accurate Bragg intensities from a heterogeneous population of microcrystals, while at the same time modeling the X-ray spectrum and detector geometry. Here we present a computational approach designed to extract statistically significant high-resolution signals from fewer diffraction measurements. PMID:24633409

  12. Aplanatic Three-Mirror Objective for High-Magnification Soft X-Ray Microscopy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Toyoda, M.; Jinno, T.; Yanagihara, M.

    2011-09-09

    An innovative solution for high-magnification microscopy, based on attaching afocal optics for focal length reduction, is proposed. The solution, consisting of three spherical mirrors, allows one to enhance a magnification of a laboratory based soft x-ray microscope over 1000x, where movies with diffraction-limited resolution can be observed with an x-ray CCD. The design example, having a numerical aperture of 0.25, was successfully demonstrated both a high magnification and a large field of view.

  13. Long-baseline optical intensity interferometry. Laboratory demonstration of diffraction-limited imaging

    NASA Astrophysics Data System (ADS)

    Dravins, Dainis; Lagadec, Tiphaine; Nuñez, Paul D.

    2015-08-01

    Context. A long-held vision has been to realize diffraction-limited optical aperture synthesis over kilometer baselines. This will enable imaging of stellar surfaces and their environments, and reveal interacting gas flows in binary systems. An opportunity is now opening up with the large telescope arrays primarily erected for measuring Cherenkov light in air induced by gamma rays. With suitable software, such telescopes could be electronically connected and also used for intensity interferometry. Second-order spatial coherence of light is obtained by cross correlating intensity fluctuations measured in different pairs of telescopes. With no optical links between them, the error budget is set by the electronic time resolution of a few nanoseconds. Corresponding light-travel distances are approximately one meter, making the method practically immune to atmospheric turbulence or optical imperfections, permitting both very long baselines and observing at short optical wavelengths. Aims: Previous theoretical modeling has shown that full images should be possible to retrieve from observations with such telescope arrays. This project aims at verifying diffraction-limited imaging experimentally with groups of detached and independent optical telescopes. Methods: In a large optics laboratory, artificial stars (single and double, round and elliptic) were observed by an array of small telescopes. Using high-speed photon-counting solid-state detectors and real-time electronics, intensity fluctuations were cross-correlated over up to 180 baselines between pairs of telescopes, producing coherence maps across the interferometric Fourier-transform plane. Results: These interferometric measurements were used to extract parameters about the simulated stars, and to reconstruct their two-dimensional images. As far as we are aware, these are the first diffraction-limited images obtained from an optical array only linked by electronic software, with no optical connections between the telescopes. Conclusions: These experiments serve to verify the concepts for long-baseline aperture synthesis in the optical, somewhat analogous to radio interferometry.

  14. A New Camera for Powder Diffraction of Macromolecular Crystallography at SPring-8

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Miura, Keiko; Inoue, Katsuaki; Goto, Shunji

    2004-05-12

    A powder diffractometer of Guinier geometry was developed and tested on a beamline, BL40B2, at SPring-8. The long specimen-to-detector distance, 1,000 mm, is advantageous in recording diffraction from Bragg spacing of 20 nm or larger. The angular resolution, 0.012 degrees, was realized together with the focusing optics, the long specimen-to-detector distance and the small pixel size of Blue-type Imaging Plate detector. Such a high resolution makes the peak separation possible in the powder diffraction from microcrystals with large unit cell and low symmetry of biological macromolecules.

  15. X-Ray Topography of Tetragonal Lysozyme Grown by the Temperature-Controlled Technique

    NASA Technical Reports Server (NTRS)

    Stojanoff, V.; Siddons, D. P.; Monaco, Lisa A.; Vekilov, Peter; Rosenberger, Franz

    1997-01-01

    Growth-induced defects in lysozyme crystals were observed by white-beam and monochromatic X-ray topography at the National Synchrotron Light Source (NSLS) at the Brookhaven National Laboratory (BNL). The topographic methods were non-destructive to the extent that traditional diffraction data collection could be performed to high resolution after topography. It was found that changes in growth parameters, defect concentration as detected by X-ray topography, and the diffraction quality obtainable from the crystals were all strongly correlated. In addition, crystals with fewer defects showed lower mosaicity and higher diffraction resolution as expected.

  16. Partially coherent X-ray wavefront propagation simulations including grazing-incidence focusing optics.

    PubMed

    Canestrari, Niccolo; Chubar, Oleg; Reininger, Ruben

    2014-09-01

    X-ray beamlines in modern synchrotron radiation sources make extensive use of grazing-incidence reflective optics, in particular Kirkpatrick-Baez elliptical mirror systems. These systems can focus the incoming X-rays down to nanometer-scale spot sizes while maintaining relatively large acceptance apertures and high flux in the focused radiation spots. In low-emittance storage rings and in free-electron lasers such systems are used with partially or even nearly fully coherent X-ray beams and often target diffraction-limited resolution. Therefore, their accurate simulation and modeling has to be performed within the framework of wave optics. Here the implementation and benchmarking of a wave-optics method for the simulation of grazing-incidence mirrors based on the local stationary-phase approximation or, in other words, the local propagation of the radiation electric field along geometrical rays, is described. The proposed method is CPU-efficient and fully compatible with the numerical methods of Fourier optics. It has been implemented in the Synchrotron Radiation Workshop (SRW) computer code and extensively tested against the geometrical ray-tracing code SHADOW. The test simulations have been performed for cases without and with diffraction at mirror apertures, including cases where the grazing-incidence mirrors can be hardly approximated by ideal lenses. Good agreement between the SRW and SHADOW simulation results is observed in the cases without diffraction. The differences between the simulation results obtained by the two codes in diffraction-dominated cases for illumination with fully or partially coherent radiation are analyzed and interpreted. The application of the new method for the simulation of wavefront propagation through a high-resolution X-ray microspectroscopy beamline at the National Synchrotron Light Source II (Brookhaven National Laboratory, USA) is demonstrated.

  17. High-resolution study of dynamical diffraction phenomena accompanying the Renninger (222/113) case of three-beam diffraction in silicon

    PubMed Central

    Kazimirov, A.; Kohn, V. G.

    2010-01-01

    X-ray optical schemes capable of producing a highly monochromatic beam with high angular collimation in both the vertical and horizontal planes have been evaluated and utilized to study high-resolution diffraction phenomena in the Renninger (222/113) case of three-beam diffraction in silicon. The effect of the total reflection of the incident beam into the nearly forbidden reflected beam was observed for the first time with the maximum 222 reflectivity at the 70% level. We have demonstrated that the width of the 222 reflection can be varied many times by tuning the azimuthal angle by only a few µrad in the vicinity of the three-beam diffraction region. This effect, predicted theoretically more than 20 years ago, is explained by the enhancement of the 222 scattering amplitude due to the virtual two-stage 000 113 222 process which depends on the azimuthal angle. PMID:20555185

  18. Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations

    NASA Astrophysics Data System (ADS)

    Schulz-Hildebrandt, H.; Münter, Michael; Ahrens, M.; Spahr, H.; Hillmann, D.; König, P.; Hüttmann, G.

    2018-03-01

    Optical coherence tomography (OCT) images scattering tissues with 5 to 15 μm resolution. This is usually not sufficient for a distinction of cellular and subcellular structures. Increasing axial and lateral resolution and compensation of artifacts caused by dispersion and aberrations is required to achieve cellular and subcellular resolution. This includes defocus which limit the usable depth of field at high lateral resolution. OCT gives access the phase of the scattered light and hence correction of dispersion and aberrations is possible by numerical algorithms. Here we present a unified dispersion/aberration correction which is based on a polynomial parameterization of the phase error and an optimization of the image quality using Shannon's entropy. For validation, a supercontinuum light sources and a costume-made spectrometer with 400 nm bandwidth were combined with a high NA microscope objective in a setup for tissue and small animal imaging. Using this setup and computation corrections, volumetric imaging at 1.5 μm resolution is possible. Cellular and near cellular resolution is demonstrated in porcine cornea and the drosophila larva, when computational correction of dispersion and aberrations is used. Due to the excellent correction of the used microscope objective, defocus was the main contribution to the aberrations. In addition, higher aberrations caused by the sample itself were successfully corrected. Dispersion and aberrations are closely related artifacts in microscopic OCT imaging. Hence they can be corrected in the same way by optimization of the image quality. This way microscopic resolution is easily achieved in OCT imaging of static biological tissues.

  19. Single Fluorescent Molecules as Nano-Illuminators for Biological Structure and Function

    NASA Astrophysics Data System (ADS)

    Moerner, W. E.

    2011-03-01

    Since the first optical detection and spectroscopy of a single molecule in a solid (Phys. Rev. Lett. {62}, 2535 (1989)), much has been learned about the ability of single molecules to probe local nanoenvironments and individual behavior in biological and nonbiological materials in the absence of ensemble averaging that can obscure heterogeneity. Because each single fluorophore acts a light source roughly 1 nm in size, microscopic imaging of individual fluorophores leads naturally to superlocalization, or determination of the position of the molecule with precision beyond the optical diffraction limit, simply by digitization of the point-spread function from the single emitter. For example, the shape of single filaments in a living cell can be extracted simply by allowing a single molecule to move through the filament (PNAS {103}, 10929 (2006)). The addition of photoinduced control of single-molecule emission allows imaging beyond the diffraction limit (super-resolution) and a new array of acronyms (PALM, STORM, F-PALM etc.) and advances have appeared. We have used the native blinking and switching of a common yellow-emitting variant of green fluorescent protein (EYFP) reported more than a decade ago (Nature {388}, 355 (1997)) to achieve sub-40 nm super-resolution imaging of several protein structures in the bacterium Caulobacter crescentus: the quasi-helix of the actin-like protein MreB (Nat. Meth. {5}, 947 (2008)), the cellular distribution of the DNA binding protein HU (submitted), and the recently discovered division spindle composed of ParA filaments (Nat. Cell Biol. {12}, 791 (2010)). Even with these advances, better emitters would provide more photons and improved resolution, and a new photoactivatable small-molecule emitter has recently been synthesized and targeted to specific structures in living cells to provide super-resolution images (JACS {132}, 15099 (2010)). Finally, a new optical method for extracting three-dimensional position information based on a double-helix point spread function enables quantitative tracking of single mRNA particles in living yeast cells with 15 ms time resolution and 25-50 nm spatial precision (PNAS {107}, 17864 (2010)). These examples illustrate the power of single-molecule optical imaging in extracting new structural and functional information in living cells.

  20. X-ray Echo Spectroscopy

    NASA Astrophysics Data System (ADS)

    Shvyd'ko, Yuri

    2016-02-01

    X-ray echo spectroscopy, a counterpart of neutron spin echo, is being introduced here to overcome limitations in spectral resolution and weak signals of the traditional inelastic x-ray scattering (IXS) probes. An image of a pointlike x-ray source is defocused by a dispersing system comprised of asymmetrically cut specially arranged Bragg diffracting crystals. The defocused image is refocused into a point (echo) in a time-reversal dispersing system. If the defocused beam is inelastically scattered from a sample, the echo signal acquires a spatial distribution, which is a map of the inelastic scattering spectrum. The spectral resolution of the echo spectroscopy does not rely on the monochromaticity of the x rays, ensuring strong signals along with a very high spectral resolution. Particular schemes of x-ray echo spectrometers for 0.1-0.02 meV ultrahigh-resolution IXS applications (resolving power >108 ) with broadband ≃5 - 13 meV dispersing systems are introduced featuring more than 103 signal enhancement. The technique is general, applicable in different photon frequency domains.

  1. Label-free photoacoustic nanoscopy

    PubMed Central

    Danielli, Amos; Maslov, Konstantin; Garcia-Uribe, Alejandro; Winkler, Amy M.; Li, Chiye; Wang, Lidai; Chen, Yun; Dorn, Gerald W.; Wang, Lihong V.

    2014-01-01

    Abstract. Super-resolution microscopy techniques—capable of overcoming the diffraction limit of light—have opened new opportunities to explore subcellular structures and dynamics not resolvable in conventional far-field microscopy. However, relying on staining with exogenous fluorescent markers, these techniques can sometimes introduce undesired artifacts to the image, mainly due to large tagging agent sizes and insufficient or variable labeling densities. By contrast, the use of endogenous pigments allows imaging of the intrinsic structures of biological samples with unaltered molecular constituents. Here, we report label-free photoacoustic (PA) nanoscopy, which is exquisitely sensitive to optical absorption, with an 88 nm resolution. At each scanning position, multiple PA signals are successively excited with increasing laser pulse energy. Because of optical saturation or nonlinear thermal expansion, the PA amplitude depends on the nonlinear incident optical fluence. The high-order dependence, quantified by polynomial fitting, provides super-resolution imaging with optical sectioning. PA nanoscopy is capable of super-resolution imaging of either fluorescent or nonfluorescent molecules. PMID:25104412

  2. Advances in high-resolution imaging--techniques for three-dimensional imaging of cellular structures.

    PubMed

    Lidke, Diane S; Lidke, Keith A

    2012-06-01

    A fundamental goal in biology is to determine how cellular organization is coupled to function. To achieve this goal, a better understanding of organelle composition and structure is needed. Although visualization of cellular organelles using fluorescence or electron microscopy (EM) has become a common tool for the cell biologist, recent advances are providing a clearer picture of the cell than ever before. In particular, advanced light-microscopy techniques are achieving resolutions below the diffraction limit and EM tomography provides high-resolution three-dimensional (3D) images of cellular structures. The ability to perform both fluorescence and electron microscopy on the same sample (correlative light and electron microscopy, CLEM) makes it possible to identify where a fluorescently labeled protein is located with respect to organelle structures visualized by EM. Here, we review the current state of the art in 3D biological imaging techniques with a focus on recent advances in electron microscopy and fluorescence super-resolution techniques.

  3. Diffraction-limited imaging with very large telescopes; Proceedings of the NATO Advanced Study Institute, Cargese, France, Sept. 13-23, 1988

    NASA Astrophysics Data System (ADS)

    Alloin, D. M.; Mariotti, J.-M.

    Recent advances in optics and observation techniques for very large astronomical telescopes are discussed in reviews and reports. Topics addressed include Fourier optics and coherence, optical propagation and image formation through a turbulent atmosphere, radio telescopes, continuously deformable telescopes for optical interferometry (I), amplitude estimation from speckle I, noise calibration of speckle imagery, and amplitude estimation from diluted-array I. Consideration is given to first-order imaging methods, speckle imaging with the PAPA detector and the Knox-Thompson algorithm, phase-closure imaging, real-time wavefront sensing and adaptive optics, differential I, astrophysical programs for high-angular-resolution optical I, cophasing telescope arrays, aperture synthesis for space observatories, and lunar occultations for marcsec resolution.

  4. Principle, system, and applications of tip-enhanced Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Zhang, MingQian; Wang, Rui; Wu, XiaoBin; Wang, Jia

    2012-08-01

    Raman spectroscopy is a powerful technique in chemical information characterization. However, this spectral method is subject to two obstacles in nano-material detection. One is diffraction limited spatial resolution, and the other is its inherent small Raman cross section and weak signaling. To resolve these problems, a new approach has been developed, denoted as tip-enhanced Raman spectroscopy (TERS). TERS is capable of high-resolution and high-sensitivity detection and demonstrated to be a promising spectroscopic and micro-topographic method to characterize nano-materials and nanostructures. In this paper, the principle and experimental system of TERS are discussed. The latest application of TERS in molecule detection, biological specimen identification, nanao-material characterization, and semi-conductor material determination with some specific experimental examples are presented.

  5. 30-lens interferometer for high energy x-rays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lyubomirskiy, M., E-mail: lyubomir@esrf.fr; Snigireva, I., E-mail: irina@esrf.fr; Vaughan, G.

    2016-07-27

    We report a hard X-ray multilens interferometer consisting of 30 parallel compound refractive lenses. Under coherent illumination each CRL creates a diffraction limited focal spot - secondary source. An overlapping of coherent beams from these sources resulting in the interference pattern which has a rich longitudinal structure in accordance with the Talbot imaging formalism. The proposed interferometer was experimentally tested at ID11 ESRF beamline for the photon energies 32 keV and 65 keV. The fundamental and fractional Talbot images were recorded with the high resolution CCD camera. An effective source size in the order of 15 µm was determined frommore » the first Talbot image proving that the multilens interferometer can be used as a high resolution beam diagnostic tool.« less

  6. Superlensing effect for surface acoustic waves in a pillar-based phononic crystal with negative refractive index

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Addouche, Mahmoud, E-mail: mamoud.addouche@femto-st.fr; Al-Lethawe, Mohammed A., E-mail: mohammed.abdulridha@femto-st.fr; Choujaa, Abdelkrim, E-mail: achoujaa@femto-st.fr

    2014-07-14

    We demonstrate super resolution imaging for surface acoustic waves using a phononic structure displaying negative refractive index. This phononic structure is made of a monolithic square lattice of cylindrical pillars standing on a semi-infinite medium. The pillars act as acoustic resonator and induce a surface propagating wave with unusual dispersion. We found, under specific geometrical parameters, one propagating mode that exhibits negative refraction effect with negative effective index close to −1. Furthermore, a flat lens with finite number of pillars is designed to allow the focusing of an acoustic point source into an image with a resolution of (λ)/3 ,more » overcoming the Rayleigh diffraction limit.« less

  7. Observational Evidence for Small-Scale Mixture of Weak and Strong Fields in the Quiet Sun

    NASA Astrophysics Data System (ADS)

    Socas-Navarro, H.; Lites, B. W.

    2004-11-01

    Three different maps of the quiet Sun, observed with the Advanced Stokes Polarimeter (ASP) and the Diffraction-Limited Stokes Polarimeter (DLSP), show evidence of strong (~=1700 G) and weak (<500 G) fields coexisting within the resolution element at both network and internetwork locations. The angular resolution of the observations is of 1" (ASP) and 0.6" (DLSP). Even at the higher DLSP resolution, a significant fraction of the network magnetic patches harbor a mixture of strong and weak fields. Internetwork elements that exhibit kG fields when analyzed with a single-component atmosphere are also shown to harbor considerable amounts of weak fields. Only those patches for which a single-component analysis yields weak fields do not show this mixture of field strengths. Finally, there is a larger fractional area of weak fields in the convective upflows than in the downflows.

  8. Design and evaluation of an ultra-slim objective for in-vivo deep optical biopsy

    PubMed Central

    Landau, Sara M.; Liang, Chen; Kester, Robert T.; Tkaczyk, Tomasz S.; Descour, Michael R.

    2010-01-01

    An estimated 1.6 million breast biopsies are performed in the US each year. In order to provide real-time, in-vivo imaging with sub-cellular resolution for optical biopsies, we have designed an ultra-slim objective to fit inside the 1-mm-diameter hypodermic needles currently used for breast biopsies to image tissue stained by the fluorescent probe proflavine. To ensure high-quality imaging performance, experimental tests were performed to characterize fiber bundle’s light-coupling efficiency and simulations were performed to evaluate the impact of candidate lens materials’ autofluorescence. A prototype of NA = 0.4, 250-µm field of view, ultra-slim objective optics was built and tested, yielding diffraction-limited performance and estimated resolution of 0.9 µm. When used in conjunction with a commercial coherent fiber bundle to relay the image formed by the objective, the measured resolution was 2.5 µm. PMID:20389489

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

    DOE PAGES

    Meddens, Marjolein B. M.; Liu, Sheng; Finnegan, Patrick S.; ...

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

    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.

  11. Near-field examination of perovskite-based superlenses and superlens-enhanced probe-object coupling.

    PubMed

    Kehr, S C; Liu, Y M; Martin, L W; Yu, P; Gajek, M; Yang, S-Y; Yang, C-H; Wenzel, M T; Jacob, R; von Ribbeck, H-G; Helm, M; Zhang, X; Eng, L M; Ramesh, R

    2011-01-01

    A planar slab of negative-index material works as a superlens with sub-diffraction-limited resolution, as propagating waves are focused and, moreover, evanescent waves are reconstructed in the image plane. Here we demonstrate a superlens for electric evanescent fields with low losses using perovskites in the mid-infrared regime. The combination of near-field microscopy with a tunable free-electron laser allows us to address precisely the polariton modes, which are critical for super-resolution imaging. We spectrally study the lateral and vertical distributions of evanescent waves around the image plane of such a lens, and achieve imaging resolution of λ/14 at the superlensing wavelength. Interestingly, at certain distances between the probe and sample surface, we observe a maximum of these evanescent fields. Comparisons with numerical simulations indicate that this maximum originates from an enhanced coupling between probe and object, which might be applicable for multifunctional circuits, infrared spectroscopy and thermal sensors.

  12. Sub-10 fs Time-Resolved Vibronic Optical Microscopy

    PubMed Central

    2016-01-01

    We introduce femtosecond wide-field transient absorption microscopy combining sub-10 fs pump and probe pulses covering the complete visible (500–650 nm) and near-infrared (650–950 nm) spectrum with diffraction-limited optical resolution. We demonstrate the capabilities of our system by reporting the spatially- and spectrally-resolved transient electronic response of MAPbI3–xClx perovskite films and reveal significant quenching of the transient bleach signal at grain boundaries. The unprecedented temporal resolution enables us to directly observe the formation of band-gap renormalization, completed in 25 fs after photoexcitation. In addition, we acquire hyperspectral Raman maps of TIPS pentacene films with sub-400 nm spatial and sub-15 cm–1 spectral resolution covering the 100–2000 cm–1 window. Our approach opens up the possibility of studying ultrafast dynamics on nanometer length and femtosecond time scales in a variety of two-dimensional and nanoscopic systems. PMID:27934055

  13. Optical gating and streaking of free electrons with sub-optical cycle precision

    PubMed Central

    Kozák, M.; McNeur, J.; Leedle, K. J.; Deng, H.; Schönenberger, N.; Ruehl, A.; Hartl, I.; Harris, J. S.; Byer, R. L.; Hommelhoff, P.

    2017-01-01

    The temporal resolution of ultrafast electron diffraction and microscopy experiments is currently limited by the available experimental techniques for the generation and characterization of electron bunches with single femtosecond or attosecond durations. Here, we present proof of principle experiments of an optical gating concept for free electrons via direct time-domain visualization of the sub-optical cycle energy and transverse momentum structure imprinted on the electron beam. We demonstrate a temporal resolution of 1.2±0.3 fs. The scheme is based on the synchronous interaction between electrons and the near-field mode of a dielectric nano-grating excited by a femtosecond laser pulse with an optical period duration of 6.5 fs. The sub-optical cycle resolution demonstrated here is promising for use in laser-driven streak cameras for attosecond temporal characterization of bunched particle beams as well as time-resolved experiments with free-electron beams. PMID:28120930

  14. Maximizing the Biochemical Resolving Power of Fluorescence Microscopy

    PubMed Central

    Esposito, Alessandro; Popleteeva, Marina; Venkitaraman, Ashok R.

    2013-01-01

    Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been formally described, because an adequate and general theoretical framework is lacking. Here, we develop a mathematical characterization of the biochemical resolution in fluorescence detection with Fisher information analysis. To improve the precision and the resolution of quantitative imaging methods, we demonstrate strategies for the optimization of fluorescence lifetime, fluorescence anisotropy and hyperspectral detection, as well as different multi-dimensional techniques. We describe optimized imaging protocols, provide optimization algorithms and describe precision and resolving power in biochemical imaging thanks to the analysis of the general properties of Fisher information in fluorescence detection. These strategies enable the optimal use of the information content available within the limited photon-budget typically available in fluorescence microscopy. This theoretical foundation leads to a generalized strategy for the optimization of multi-dimensional optical detection, and demonstrates how the parallel detection of all properties of fluorescence can maximize the biochemical resolving power of fluorescence microscopy, an approach we term Hyper Dimensional Imaging Microscopy (HDIM). Our work provides a theoretical framework for the description of the biochemical resolution in fluorescence microscopy, irrespective of spatial resolution, and for the development of a new class of microscopes that exploit multi-parametric detection systems. PMID:24204821

  15. A preliminary neutron diffraction study of rasburicase, a recombinant urate oxidase enzyme, complexed with 8-azaxanthin

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Budayova-Spano, Monika, E-mail: spano@embl-grenoble.fr; Institut Laue-Langevin, 6 Rue Jules Horowitz, BP 156, 38042 Grenoble; Bonneté, Françoise

    2006-03-01

    Neutron diffraction data of hydrogenated recombinant urate oxidase enzyme (Rasburicase), complexed with a purine-type inhibitor 8-azaxanthin, was collected to 2.1 Å resolution from a crystal grown in D{sub 2}O by careful control and optimization of crystallization conditions via knowledge of the phase diagram. Deuterium atoms were clearly seen in the neutron-scattering density map. Crystallization and preliminary neutron diffraction measurements of rasburicase, a recombinant urate oxidase enzyme expressed by a genetically modified Saccharomyces cerevisiae strain, complexed with a purine-type inhibitor (8-azaxanthin) are reported. Neutron Laue diffraction data were collected to 2.1 Å resolution using the LADI instrument from a crystal (grownmore » in D{sub 2}O) with volume 1.8 mm{sup 3}. The aim of this neutron diffraction study is to determine the protonation states of the inhibitor and residues within the active site. This will lead to improved comprehension of the enzymatic mechanism of this important enzyme, which is used as a protein drug to reduce toxic uric acid accumulation during chemotherapy. This paper illustrates the high quality of the neutron diffraction data collected, which are suitable for high-resolution structural analysis. In comparison with other neutron protein crystallography studies to date in which a hydrogenated protein has been used, the volume of the crystal was relatively small and yet the data still extend to high resolution. Furthermore, urate oxidase has one of the largest primitive unit-cell volumes (space group I222, unit-cell parameters a = 80, b = 96, c = 106 Å) and molecular weights (135 kDa for the homotetramer) so far successfully studied with neutrons.« less

  16. Biological imaging by soft x-ray diffraction microscopy

    DOE PAGES

    Shapiro, D.; Thibault, P.; Beetz, T.; ...

    2005-10-25

    We have used the method of x-ray diffraction microscopy to image the complex-valued exit wave of an intact and unstained yeast cell. The images of the freeze-dried cell, obtained by using 750-eV x-rays from different angular orientations, portray several of the cell's major internal components to 30-nm resolution. The good agreement among the independently recovered structures demonstrates the accuracy of the imaging technique. To obtain the best possible reconstructions, we have implemented procedures for handling noisy and incomplete diffraction data, and we propose a method for determining the reconstructed resolution. This work represents a previously uncharacterized application of x-ray diffractionmore » microscopy to a specimen of this complexity and provides confidence in the feasibility of the ultimate goal of imaging biological specimens at 10-nm resolution in three dimensions.« less

  17. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Aoki, Ken-ichi; Tanaka, Nobutada, E-mail: ntanaka@pharm.showa-u.ac.jp; Ishikura, Shuhei

    Pig heart carbonyl reductase has been crystallized in the presence of NADPH. Diffraction data have been collected using synchrotron radiation. Pig heart carbonyl reductase (PHCR), which belongs to the short-chain dehydrogenase/reductase (SDR) family, has been crystallized by the hanging-drop vapour-diffusion method. Two crystal forms (I and II) have been obtained in the presence of NADPH. Form I crystals belong to the tetragonal space group P4{sub 2}, with unit-cell parameters a = b = 109.61, c = 94.31 Å, and diffract to 1.5 Å resolution. Form II crystals belong to the tetragonal space group P4{sub 1}2{sub 1}2, with unit-cell parameters amore » = b = 120.10, c = 147.00 Å, and diffract to 2.2 Å resolution. Both crystal forms are suitable for X-ray structure analysis at high resolution.« less

  18. A Compact "Water Window" Microscope with 60 nm Spatial Resolution for Applications in Biology and Nanotechnology.

    PubMed

    Wachulak, Przemyslaw; Torrisi, Alfio; Nawaz, Muhammad F; Bartnik, Andrzej; Adjei, Daniel; Vondrová, Šárka; Turňová, Jana; Jančarek, Alexandr; Limpouch, Jiří; Vrbová, Miroslava; Fiedorowicz, Henryk

    2015-10-01

    Short illumination wavelength allows an extension of the diffraction limit toward nanometer scale; thus, improving spatial resolution in optical systems. Soft X-ray (SXR) radiation, from "water window" spectral range, λ=2.3-4.4 nm wavelength, which is particularly suitable for biological imaging due to natural optical contrast provides better spatial resolution than one obtained with visible light microscopes. The high contrast in the "water window" is obtained because of selective radiation absorption by carbon and water, which are constituents of the biological samples. The development of SXR microscopes permits the visualization of features on the nanometer scale, but often with a tradeoff, which can be seen between the exposure time and the size and complexity of the microscopes. Thus, herein, we present a desk-top system, which overcomes the already mentioned limitations and is capable of resolving 60 nm features with very short exposure time. Even though the system is in its initial stage of development, we present different applications of the system for biology and nanotechnology. Construction of the microscope with recently acquired images of various samples will be presented and discussed. Such a high resolution imaging system represents an interesting solution for biomedical, material science, and nanotechnology applications.

  19. Inquiry with Laser Printer Diffraction Gratings

    ERIC Educational Resources Information Center

    Van Hook, Stephen J.

    2007-01-01

    The pages of "The Physics Teacher" have featured several clever designs for homemade diffraction gratings using a variety of materials--cloth, lithographic film, wire, compact discs, parts of aerosol spray cans, and pseudoliquids and pseudosolids. A different and inexpensive method I use to make low-resolution diffraction gratings takes advantage…

  20. A general strategy for developing cell-permeable photo-modulatable organic fluorescent probes for live-cell super-resolution imaging.

    PubMed

    Pan, Deng; Hu, Zhe; Qiu, Fengwu; Huang, Zhen-Li; Ma, Yilong; Wang, Yina; Qin, Lingsong; Zhang, Zhihong; Zeng, Shaoqun; Zhang, Yu-Hui

    2014-11-20

    Single-molecule localization microscopy (SMLM) achieves super-resolution imaging beyond the diffraction limit but critically relies on the use of photo-modulatable fluorescent probes. Here we report a general strategy for constructing cell-permeable photo-modulatable organic fluorescent probes for live-cell SMLM by exploiting the remarkable cytosolic delivery ability of a cell-penetrating peptide (rR)3R2. We develop photo-modulatable organic fluorescent probes consisting of a (rR)3R2 peptide coupled to a cell-impermeable organic fluorophore and a recognition unit. Our results indicate that these organic probes are not only cell permeable but can also specifically and directly label endogenous targeted proteins. Using the probes, we obtain super-resolution images of lysosomes and endogenous F-actin under physiological conditions. We resolve the dynamics of F-actin with 10 s temporal resolution in live cells and discern fine F-actin structures with diameters of ~80 nm. These results open up new avenues in the design of fluorescent probes for live-cell super-resolution imaging.

  1. 120nm resolution in thick samples with structured illumination and adaptive optics

    NASA Astrophysics Data System (ADS)

    Thomas, Benjamin; Sloan, Megan; Wolstenholme, Adrian J.; Kner, Peter

    2014-03-01

    μLinear Structured Illumination Microscopy (SIM) provides a two-fold increase over the diffraction limited resolution. SIM produces excellent images with 120nm resolution in tissue culture cells in two and three dimensions. For SIM to work correctly, the point spread function (PSF) and optical transfer function (OTF) must be known, and, ideally, should be unaberrated. When imaging through thick samples, aberrations will be introduced into the optical system which will reduce the peak intensity and increase the width of the PSF. This will lead to reduced resolution and artifacts in SIM images. Adaptive optics can be used to correct the optical wavefront restoring the PSF to its unaberrated state, and AO has been used in several types of fluorescence microscopy. We demonstrate that AO can be used with SIM to achieve 120nm resolution through 25m of tissue by imaging through the full thickness of an adult C. elegans roundworm. The aberrations can be corrected over a 25μm × 45μm field of view with one wavefront correction setting, demonstrating that AO can be used effectively with widefield superresolution techniques.

  2. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhou, Dongwen; Sun, Jianping; Zhao, Wei

    The CRD domain of GRP from H. sapiens has been expressed, purified and crystallized and X-ray diffraction data have been collected to a resolution of 2.0 Å. Galectins are a family of animal lectins which share similar carbohydrate-recognition domains (CRDs) and an affinity for β-galactosides. A novel human galectin-related protein named GRP (galectin-related protein; previously known as HSPC159) comprises only one conserved CRD with 38 additional N-terminal residues. The C-terminal fragment of human GRP (GRP-C; residues 38–172) containing the CRD has been expressed and purified. The protein was crystallized using the hanging-drop vapour-diffusion method from a solution containing 2% PEGmore » 400 and 2M ammonium sulfate in 100 mM Tris–HCl buffer pH 7.5. Diffraction data were collected to a resolution limit of 2.0 Å at beamline 3W1A of Beijing Synchrotron Radiation Facility at 100 K. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 123.07, b = 96.67, c = 61.56 Å, β = 118.72°. The estimated Matthews coefficient was 2.6 Å{sup 3} Da{sup −1}, corresponding to 51.8% solvent content.« less

  3. RIMAS - Optical Design Development of the Imager/Spectrometer for the Discovery Channel Telescope

    NASA Technical Reports Server (NTRS)

    Capone, John

    2012-01-01

    The Rapid IMAger - Spectrometer (RIMAS) is a collaborative effort between the University of Maryland at College Park, NASA-GSFC and Lowell Observatory designed for use on the 4.3 meter Discovery Channel Telescope at Lowell. The primary science goal of the instrument is the study of gamma-ray burst (ORB) afterglow appearing in the near-infrared. Continuous operation will allow measurements beginning minutes after the prompt emission. We present the results of the RIMAS optical design development. The instrument consists of two arms separated by a dichroic: the first for the Y and J bands (0.9 - 1.35 microns) and the second for the Hand K-bands (1.5 - 1.8 and 2.0 - 2.4 microns). Each arm will be equipped with two broad band filters for imaging, as well as low resolution and echelle grisms. The imaging modes are designed to be diffraction limited, with one pixel corresponding to approx.0.35 arcseconds, while the diffractive modes have resolving powers of approximately 20 and 4,000. With photometric and spectroscopic capabilities, RIMAS will be well positioned to quickly determine redshifts, followed by high resolution spectroscopic studies of ORB afterglow.

  4. Ultrahigh resolution protein crystallography: Concanavalin A to 0.94 Å and beyond

    NASA Astrophysics Data System (ADS)

    Deacon, A. M.; Gleichmann, T.; Harrop, S. J.; Helliwell, J. R.; Kalb Gilboa, A. J.; Yariv, J.

    1996-09-01

    Many years ago the idea of collecting voluminous quantities of weak reflection intensities from a protein crystal, at high resolution, was a particular challenge [J.R. Helliwell (1979) Daresbury Study Weekend DL/SCI R13, pp. 1-6]. The combination of insertion devices with very high x-ray fluxes at short x-ray wavelengths, sensitive CCD detectors, and freezing of crystals have provided the means to certainly match those best hopes. So much so that the data can best be described as ultrahigh resolution, at least as evidenced in our studies of the 25000 molecular weight plant protein concanavalin A. (The intrinsic property of this protein is to bind sugar molecules; it is implicated in cell-to-cell recognition processes and is widely used as a laboratory diagnostic tool.) At CHESS we have used a 0.9 Å wavelength beam on station A1, fed by a 24 pole multipole wiggler. Both an imaging plate system and the Princeton 1k CCD detector [M. Tate et al., J. Appl. Cryst. 28, 196 (1995)] have been used on this experimental setup to collect diffraction data sets from frozen concanavalin A crystals (saccharide-free crystal form). The rapid readout of the CCD was most convenient compared with the image plate and its associated scanning and erasing. Moreover the data processing results towards the edges of the detectors, 0.98 Å, show that the CCD is much better than the image plate at recording these weaker data (Rmerge(I) 13% versus 44%, respectively). The poor performance of the image plate with weak signals has of course been documented by the Daresbury detector group [R. Lewis, J. Synchrotron Radiation 1, 43 (1994)]. However, the aperture of the CCD used was limiting here. Very recently, in another run at CHESS with the CCD on A1, we have been able to record diffraction data to 0.94 Å by further offsetting the detector. We again found that the reflections are still strong at the edge. Clearly the use of even shorter wavelengths than 0.9 Å would be very useful in matching the solid angle of the diffraction pattern to the available detector aperture, for a reasonable crystal-to-detector distance. In addition, absorption errors in the data can be simultaneously removed by such a strategy. Indeed, finely focused x-ray beams of, say 0.5 Å wavelength, are especially well suited to high energy, low emittance synchrotron radition (SR) machines. Some initial tests carried out on CHESS station F2 with a 0.5 Å wavelength beam and the CCD detector show an improvement in the R-merge(I) to 2 Å resolution, in comparison to the data collected at 0.9 Å wavelength (i.e., 2.3% versus 3.0%). In conclusion, the diffraction resolution limit (0.94 Å) seen already in our concanavalin A studies can be further enhanced and is important for the most detailed molecular model refinement (and the testing of structure solving strategies), in conjunction with novel spectroscopic and theoretical studies. This paper builds upon the work of Deacon et al. [Rev. Sci. Instrum. 66, 1287 (1995)].

  5. Exploring the atomic structure and conformational flexibility of a 320 Å long engineered viral fiber using X-ray crystallography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bhardwaj, Anshul; Casjens, Sherwood R.; Cingolani, Gino, E-mail: gino.cingolani@jefferson.edu

    2014-02-01

    This study presents the crystal structure of a ∼320 Å long protein fiber generated by in-frame extension of its repeated helical coiled-coil core. Protein fibers are widespread in nature, but only a limited number of high-resolution structures have been determined experimentally. Unlike globular proteins, fibers are usually recalcitrant to form three-dimensional crystals, preventing single-crystal X-ray diffraction analysis. In the absence of three-dimensional crystals, X-ray fiber diffraction is a powerful tool to determine the internal symmetry of a fiber, but it rarely yields atomic resolution structural information on complex protein fibers. An 85-residue-long minimal coiled-coil repeat unit (MiCRU) was previously identifiedmore » in the trimeric helical core of tail needle gp26, a fibrous protein emanating from the tail apparatus of the bacteriophage P22 virion. Here, evidence is provided that an MiCRU can be inserted in frame inside the gp26 helical core to generate a rationally extended fiber (gp26-2M) which, like gp26, retains a trimeric quaternary structure in solution. The 2.7 Å resolution crystal structure of this engineered fiber, which measures ∼320 Å in length and is only 20–35 Å wide, was determined. This structure, the longest for a trimeric protein fiber to be determined to such a high resolution, reveals the architecture of 22 consecutive trimerization heptads and provides a framework to decipher the structural determinants for protein fiber assembly, stability and flexibility.« less

  6. Wavefront correction and high-resolution in vivo OCT imaging with an objective integrated multi-actuator adaptive lens

    PubMed Central

    Bonora, Stefano; Jian, Yifan; Zhang, Pengfei; Zam, Azhar; Pugh, Edward N.; Zawadzki, Robert J.; Sarunic, Marinko V.

    2015-01-01

    Adaptive optics is rapidly transforming microscopy and high-resolution ophthalmic imaging. The adaptive elements commonly used to control optical wavefronts are liquid crystal spatial light modulators and deformable mirrors. We introduce a novel Multi-actuator Adaptive Lens that can correct aberrations to high order, and which has the potential to increase the spread of adaptive optics to many new applications by simplifying its integration with existing systems. Our method combines an adaptive lens with an imaged-based optimization control that allows the correction of images to the diffraction limit, and provides a reduction of hardware complexity with respect to existing state-of-the-art adaptive optics systems. The Multi-actuator Adaptive Lens design that we present can correct wavefront aberrations up to the 4th order of the Zernike polynomial characterization. The performance of the Multi-actuator Adaptive Lens is demonstrated in a wide field microscope, using a Shack-Hartmann wavefront sensor for closed loop control. The Multi-actuator Adaptive Lens and image-based wavefront-sensorless control were also integrated into the objective of a Fourier Domain Optical Coherence Tomography system for in vivo imaging of mouse retinal structures. The experimental results demonstrate that the insertion of the Multi-actuator Objective Lens can generate arbitrary wavefronts to correct aberrations down to the diffraction limit, and can be easily integrated into optical systems to improve the quality of aberrated images. PMID:26368169

  7. Coherent Diffractive Imaging: From Nanometric Down to Picometric Resolution

    NASA Astrophysics Data System (ADS)

    De Caro, Liberato; Carlino, Elvio; Siliqi, Dritan; Giannini, Cinzia

    Coherent diffractive imaging (CDI) is a novel technique for inspecting (crystalline and non-crystalline) matter from nanometric down to picometric resolution. It was used originally with X-rays and, more recently, with electrons (so-called electron diffractive imaging, or EDI). This chapter introduces basic concepts concerning CDI and addresses the different types of X-ray CDI experiments that have been conducted, namely plane wave CDI from isolated objects in forward scattering, focused-beam Fresnel CDI from isolated objects in forward scattering, Bragg CDI from nanocrystals, and keyhole CDI and ptychography from extended objects. A CDI experiment with a transmission electron microscope, alternatively named an EDI experiment, is also introduced.

  8. An Efficient Image Recovery Algorithm for Diffraction Tomography Systems

    NASA Technical Reports Server (NTRS)

    Jin, Michael Y.

    1993-01-01

    A diffraction tomography system has potential application in ultrasonic medical imaging area. It is capable of achieving imagery with the ultimate resolution of one quarter the wavelength by collecting ultrasonic backscattering data from a circular array of sensors and reconstructing the object reflectivity using a digital image recovery algorithm performed by a computer. One advantage of such a system is that is allows a relatively lower frequency wave to penetrate more deeply into the object and still achieve imagery with a reasonable resolution. An efficient image recovery algorithm for the diffraction tomography system was originally developed for processing a wide beam spaceborne SAR data...

  9. Low Copy Numbers of DC-SIGN in Cell Membrane Microdomains: Implications for Structure and Function

    PubMed Central

    Liu, Ping; Wang, Xiang; Itano, Michelle S.; Neumann, Aaron K.; de Silva, Aravinda M.; Jacobson, Ken; Thompson, Nancy L.

    2014-01-01

    Presently, there are few estimates of the number of molecules occupying membrane domains. Using a total internal reflection fluorescence microscopy (TIRFM) imaging approach, based on comparing the intensities of fluorescently labeled microdomains with those of single fluorophores, we measured the occupancy of DC-SIGN, a C-type lectin, in membrane microdomains. DC-SIGN or its mutants were labeled with primary monoclonal antibodies (mAbs) in either dendritic cells (DCs) or NIH3T3 cells, or expressed as GFP fusions in NIH3T3 cells. The number of DC-SIGN molecules per microdomain ranges from only a few to over 20, while microdomain dimensions range from the diffraction limit to > 1μm. The largest fraction of microdomains, appearing at the diffraction limit, in either immature DCs or 3T3 cells contains only 4-8 molecules of DC-SIGN, consistent with our preliminary super-resolution Blink microscopy estimates. We further show that these small assemblies are sufficient to bind and efficiently internalize a small (~50nm) pathogen, dengue virus, leading to infection of host cells. PMID:24313910

  10. Measuring a Precise Ultra-Lightweight Spaceflight Mirror on Earth: The Analysis of the SHARPI PM Mirror Figure Data during Mirror Processing at GSFC

    NASA Technical Reports Server (NTRS)

    Antonille, Scott; Content, David; Rabin, Douglas; Wallace, Thomas; Wake, Shane

    2007-01-01

    The SHARPI (Solar High Angular Resolution Photometric Imager) primary mirror is a 5kg, 0.5m paraboloid, diffraction limited at FUV wavelengths when placed in a 0-G environment. The ULE sandwich honeycomb mirror and the attached mount pads were delivered by ITT (then Kodak) in 2003 to NASA s Goddard Space Flight Center (GSFC). At GSFC, we accepted, coated, mounted, and vibration tested this mirror in preparation for flight on the PICTURES (Planet Imaging Concept Testbed Using a Rocket Experiment) mission. At each step, the integrated analysis of interferometer data and FEA models was essential to quantify the 0-G mirror figure. This task required separating nanometer sized variations from hundreds of nanometers of gravity induced distortion. The ability to isolate such features allowed in-situ monitoring of mirror figure, diagnosis of perturbations, and remediation of process errors. In this paper, we describe the technical approach used to achieve these measurements and overcome the various difficulties maintaining UV diffraction-limited performance with this aggressively lightweighted mirror.

  11. Nonlinear photothermal Mid-Infrared Microspectroscopy with Superresolution

    NASA Astrophysics Data System (ADS)

    Erramilli, Shyamsunder; Mertiri, Alket; Liu, Hui; Totachawattana, Atcha; Hong, Mi; Sander, Michelle

    2015-03-01

    We describe a nonlinear method for breaking the diffraction limit in mid-infrared microscopy using nonlinear photothermal microspectroscopy. A Quantum Cascade Laser (QCL) tuned to an infrared active vibrational molecular normal mode is used as the pump laser. A low-phase noise Erbium-doped fiber (EDFL) laser is used as the probe. When the incident intensity of the mid-infrared pump laser is increased past a critical threshold, a nanobubble is nucleated, strongly modulating the scatter of the probe beam, in agreement with prior work. Remarkably, we have also found that the photothermal spectral signature of the mid-infrared absorption bifurcates and is strongly narrowed, consistent with an effective ``mean-field'' theory of the observed pitchfork bifurcation. This ultrasharp narrowing can be exploited to obtain mid-infrared images with a resolution that breaks the diffraction limit, without the need of mechanical scanning near-field probes. The method provides a powerful new tool for hyperspectral label-free mid-infrared imaging and characterization of biological tissues and materials science and engineering. We thank our collaborators H. Altug, L. D. Ziegler, J. Mertz, for their advice and generous loan of equipment.

  12. Innovative diffraction gratings for high-resolution resonant inelastic soft x-ray scattering spectroscopy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Voronov, D.L.; Warwick, T.; Gullikson, E. M.

    2016-07-27

    High-resolution Resonant Inelastic X-ray Scattering (RIXS) requires diffraction gratings with very exacting characteristics. The gratings should provide both very high dispersion and high efficiency which are conflicting requirements and extremely challenging to satisfy in the soft x-ray region for a traditional grazing incidence geometry. To achieve high dispersion one should increase the groove density of a grating; this however results in a diffraction angle beyond the critical angle range and results in drastic efficiency loss. The problem can be solved by use of multilayer coated blazed gratings (MBG). In this work we have investigated the diffraction characteristics of MBGs viamore » numerical simulations and have developed a procedure for optimization of grating design for a multiplexed high resolution imaging spectrometer for RIXS spectroscopy to be built in sector 6 at the Advanced Light Source (ALS). We found that highest diffraction efficiency can be achieved for gratings optimized for 4{sup th} or 5{sup th} order operation. Fabrication of such gratings is an extremely challenging technological problem. We present a first experimental prototype of these gratings and report its performance. High order and high line density gratings have the potential to be a revolutionary new optical element that should have great impact in the area of soft x-ray RIXS.« less

  13. Ab initio structure determination from prion nanocrystals at atomic resolution by MicroED

    PubMed Central

    Sawaya, Michael R.; Rodriguez, Jose; Cascio, Duilio; Collazo, Michael J.; Shi, Dan; Reyes, Francis E.; Gonen, Tamir; Eisenberg, David S.

    2016-01-01

    Electrons, because of their strong interaction with matter, produce high-resolution diffraction patterns from tiny 3D crystals only a few hundred nanometers thick in a frozen-hydrated state. This discovery offers the prospect of facile structure determination of complex biological macromolecules, which cannot be coaxed to form crystals large enough for conventional crystallography or cannot easily be produced in sufficient quantities. Two potential obstacles stand in the way. The first is a phenomenon known as dynamical scattering, in which multiple scattering events scramble the recorded electron diffraction intensities so that they are no longer informative of the crystallized molecule. The second obstacle is the lack of a proven means of de novo phase determination, as is required if the molecule crystallized is insufficiently similar to one that has been previously determined. We show with four structures of the amyloid core of the Sup35 prion protein that, if the diffraction resolution is high enough, sufficiently accurate phases can be obtained by direct methods with the cryo-EM method microelectron diffraction (MicroED), just as in X-ray diffraction. The success of these four experiments dispels the concern that dynamical scattering is an obstacle to ab initio phasing by MicroED and suggests that structures of novel macromolecules can also be determined by direct methods. PMID:27647903

  14. Energy research with neutrons (ErwiN) and installation of a fast neutron powder diffraction option at the MLZ, Germany1

    PubMed Central

    Mühlbauer, Martin J.

    2018-01-01

    The need for rapid data collection and studies of small sample volumes in the range of cubic millimetres are the main driving forces for the concept of a new high-throughput monochromatic diffraction instrument at the Heinz Maier-Leibnitz Zentrum (MLZ), Germany. A large region of reciprocal space will be accessed by a detector with sufficient dynamic range and microsecond time resolution, while allowing for a variety of complementary sample environments. The medium-resolution neutron powder diffraction option for ‘energy research with neutrons’ (ErwiN) at the high-flux FRM II neutron source at the MLZ is foreseen to meet future demand. ErwiN will address studies of energy-related systems and materials with respect to their structure and uniformity by means of bulk and spatially resolved neutron powder diffraction. A set of experimental options will be implemented, enabling time-resolved studies, rapid parametric measurements as a function of external parameters and studies of small samples using an adapted radial collimator. The proposed powder diffraction option ErwiN will bridge the gap in functionality between the high-resolution powder diffractometer SPODI and the time-of-flight diffractometers POWTEX and SAPHiR at the MLZ. PMID:29896055

  15. Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals

    DOE PAGES

    Dao, E. Han; Sierra, Raymond G.; Laksmono, Hartawan; ...

    2015-04-30

    In this work, we collected radiation-damage-free data from a set of cryo-cooled crystals for a novel 30S ribosomal subunit mutant using goniometer-based femtosecond crystallography. Crystal quality assessment for these samples was conducted at the X-ray Pump Probe end-station of the Linac Coherent Light Source (LCLS) using recently introduced goniometer-based instrumentation. These 30S subunit crystals were genetically engineered to omit a 26-residue protein, Thx, which is present in the wild-type Thermus thermophilus 30S ribosomal subunit. We are primarily interested in elucidating the contribution of this ribosomal protein to the overall 30S subunit structure. To assess the viability of this study, femtosecondmore » X-ray diffraction patterns from these crystals were recorded at the LCLS during a protein crystal screening beam time. During our data collection, we successfully observed diffraction from these difficult-to-grow 30S ribosomal subunit crystals. Most of our crystals were found to diffract to low resolution, while one crystal diffracted to 3.2 Å resolution. These data suggest the feasibility of pursuing high-resolution data collection as well as the need to improve sample preparation and handling in order to collect a complete radiation-damage-free data set using an X-ray Free Electron Laser.« less

  16. Flexible filamentous virus structure from fiber diffraction

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Stubbs, Gerald; Kendall, Amy; McDonald, Michele

    2008-10-24

    Fiber diffraction data have been obtained from Narcissus mosaic virus, a potexvirus from the family Flexiviridae, and soybean mosaic virus (SMV), a potyvirus from the family Potyviridae. Analysis of the data in conjunction with cryo-electron microscopy data allowed us to determine the symmetry of the viruses and to make reconstructions of SMV at 19 {angstrom} resolution and of another potexvirus, papaya mosaic virus, at 18 {angstrom} resolution. These data include the first well-ordered data ever obtained for the potyviruses and the best-ordered data from the potexviruses, and offer the promise of eventual high resolution structure determinations.

  17. High-resolution neutron-diffraction measurements to 8 kbar

    NASA Astrophysics Data System (ADS)

    Bull, C. L.; Fortes, A. D.; Ridley, C. J.; Wood, I. G.; Dobson, D. P.; Funnell, N. P.; Gibbs, A. S.; Goodway, C. M.; Sadykov, R.; Knight, K. S.

    2017-10-01

    We describe the capability to measure high-resolution neutron powder diffraction data to a pressure of at least 8 kbar. We have used the HRPD instrument at the ISIS neutron source and a piston-cylinder design of pressure cell machined from a null-scattering titanium zirconium alloy. Data were collected under hydrostatic conditions from an elpasolite perovskite La?NiMnO?; by virtue of a thinner cell wall on the incident-beam side of the cell, it was possible to obtain data in the instrument's highest resolution back-scattering detector banks up to a maximum pressure of 8.5 kbar.

  18. X-ray ptychography

    NASA Astrophysics Data System (ADS)

    Pfeiffer, Franz

    2018-01-01

    X-ray ptychographic microscopy combines the advantages of raster scanning X-ray microscopy with the more recently developed techniques of coherent diffraction imaging. It is limited neither by the fabricational challenges associated with X-ray optics nor by the requirements of isolated specimen preparation, and offers in principle wavelength-limited resolution, as well as stable access and solution to the phase problem. In this Review, we discuss the basic principles of X-ray ptychography and summarize the main milestones in the evolution of X-ray ptychographic microscopy and tomography over the past ten years, since its first demonstration with X-rays. We also highlight the potential for applications in the life and materials sciences, and discuss the latest advanced concepts and probable future developments.

  19. High resolution imaging

    NASA Astrophysics Data System (ADS)

    Goody, R.; Papaliolios, C.; Beletic, J.

    1986-09-01

    Diffraction-limited telescopic observations were obtained of solar system objects and a program of research into Uranus, Neptune, Pluto and the asteroids based upon the data obtained was persued. Two camera systems appropriate for this work were developed. Most importantly, the PAPA photon address camera was developed and proven. Algorithms were developed for both phase and amplitude recovery and were validated on theoretical and laboratory data and to a limited extent on telescopic data. A laboratory simulator was constructed that was used for development but is also available for controlled investigation of image reconstruction. During 1985 two successful expeditions were made to Hawaii and Cerro Tololo and a large body of data on Pluto, Uranus, Neptune and two asteroids are on tape.

  20. High resolution imaging

    NASA Technical Reports Server (NTRS)

    Goody, R.; Papaliolios, C.; Beletic, J.

    1986-01-01

    Diffraction-limited telescopic observations were obtained of solar system objects and a program of research into Uranus, Neptune, Pluto and the asteroids based upon the data obtained was persued. Two camera systems appropriate for this work were developed. Most importantly, the PAPA photon address camera was developed and proven. Algorithms were developed for both phase and amplitude recovery and were validated on theoretical and laboratory data and to a limited extent on telescopic data. A laboratory simulator was constructed that was used for development but is also available for controlled investigation of image reconstruction. During 1985 two successful expeditions were made to Hawaii and Cerro Tololo and a large body of data on Pluto, Uranus, Neptune and two asteroids are on tape.

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